US Pat. No. 9,046,539

LIPOPROTEIN ANALYSIS BY DIFFERENTIAL CHARGED-PARTICLE MOBILITY

QUEST DIAGNOSTICS INVESTM...

1. A method for purifying lipoproteins for differential charged particle mobility analysis, said method comprising:
a) incubating a solution comprising lipoproteins, non-lipoproteins, dextran sulfate and a solid support comprising a paramagnetic
particle, under conditions for said lipoproteins to bind to said solid support;

b) isolating said solid support from the solution thereby separating said lipoproteins from said non-lipoproteins; and
c) releasing said lipoproteins from said solid support, wherein said released lipoproteins are suitable for differential charged
particle mobility analysis that determines size distribution of said lipoproteins;

and d) subjecting said lipoproteins to differential charged particle mobility analysis
wherein:
said lipoproteins comprise HDL and one or more selected from the group consisting of LDL, Lp(a), IDL and VLDL; and
said method does not include centrifugation.
US Pat. No. 9,187,788

METHODS FOR DETECTING GENE DYSREGULATION BY INTRAGENIC DIFFERENTIAL EXPRESSION

QUEST DIAGNOSTICS INVESTM...

1. A method for diagnosing acute myeloid leukemia (AML) or a susceptibility to AML in a subject comprising:
(a) amplifying a 5? region of a target gene transcript, if present, in a biological sample with one or more 5? target primer
pairs which are complementary to the 5? region of the target gene;

(b) amplifying a 3? region of the target gene transcript, if present, in the biological sample with one or more 3? target
primer pairs which are complementary to the 3? region of the target gene;

(c) detecting the amounts of amplification products produced by the one or more 5? target primer pairs and the one or more
3? target primer pairs;

(d) comparing the relative expression of the 5? region to the 3? region of the target gene in the biological sample to the
relative expression of the 5? region to the 3? region of the target gene in a reference sample to determine if the target
gene is dysregulated; and

(e) diagnosing the subject as having AML or a susceptibility to AML when the comparison of step (d) indicates that the target
gene is dysregulated.

US Pat. No. 9,594,074

C PEPTIDE DETECTION BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of C peptide in a sample by tandem mass spectrometry, the method comprising:
(a) subjecting a sample suspected of containing C peptide to high performance liquid chromatography (HPLC) to obtain a fraction
enriched in C peptide;

(b) subjecting the enriched C peptide to an ionization source under conditions suitable to generate one or more C peptide
ions detectable by mass spectrometry;

(c) determining the amount of one or more C peptide ions by tandem mass spectrometry;
wherein the amount of ions determined in step (c) is related to the amount of a C peptide in said sample.
US Pat. No. 9,228,237

CYSTIC FIBROSIS GENE MUTATIONS

QUEST DIAGNOSTICS INVESTM...

1. A method of detecting a deletion mutant cystic fibrosis transmembrane (CFTR) nucleic acid in an individual, comprising:
(a) contacting a biological sample comprising a CFTR nucleic acid from an individual with a detectably labeled nucleic acid
probe that specifically hybridizes to a mutant CFTR nucleic acid comprising the deletion mutation but not to a wild-type CFTR
nucleic acid; and the probe comprises the deletion mutation; and

(b) detecting the CFTR deletion mutation in the individual when a hybrid is formed between the detectably labeled nucleic
acid probe and the mutant CFTR nucleic acid,

wherein the deletion mutation is selected from the group consisting of c.2817—2821del and c. 1066—1071 del.

US Pat. No. 9,200,331

METHODS FOR THE DIAGNOSIS OF BACTERIAL VAGINOSIS

QUEST DIAGNOSTICS INVESTM...

1. A method for identifying bacterial vaginosis in a human female subject, the method comprising:
(a) obtaining a vaginal swab sample from the subject;
(b) measuring levels of Atopobium vaginae, Megasphaera genus and one or more Lactobacilli species selected from the group consisting of Lactobacillus acidophilus, Lactobacillus crispatus, and Lactobacillus jensenii in the sample, wherein the levels of Atopobium vaginae, Megasphaera genus and of one or more Lactobacilli species are detected with one or more oligonucleotides capable of specifically hybridizing to a target nucleic acid sequence
from Atopobium vaginae, Megasphaera genus, Lactobacillus acidophilus, Lactobacillus crispatus, and/or Lactobacillus jensenii, wherein levels of Gardnerella vaginalis and Mobiluncus sp are not measured; wherein the levels are determined with one or more oligonucleotide primers selected from SEQ ID NOs:
1, 2, 4, 5, 11, 12, 14 and 15, and complements thereof;

(c) calculating a diagnostic score as the ratio of a logarithmic function of the levels of the one or more Lactobacilli species and a logarithmic function of the levels of the Atopobium vaginae and Megasphaera genus; and

(d) identifying the subject as having bacterial vaginosis with the diagnostic score lower than a reference score.
US Pat. No. 9,651,556

APTAMERS AND DIAGNOSTIC METHODS FOR DETECTING THE EGF RECEPTOR

QUEST DIAGNOSTICS INVESTM...

1. A method for determining the amount of free Epidermal Growth Factor Receptor (EGFR) in a sample comprising:
(a) contacting a sample comprising EGFR with an aptamer that specifically binds to free EGFR under conditions that allow binding
of the aptamer to the free EGFR in the sample to form an aptamer-EGFR complex, wherein the free EGFR is EGFR that is not bound
to a therapeutic molecule if present in the sample, and wherein the aptamer comprises the structure defined by SEQ ID NO:
55;

(b) detecting the amount of the free EGFR bound by the aptamer; and
(c) determining the amount of free EGFR in the sample based on the amount of the free EGFR bound by the aptamer.

US Pat. No. 9,422,606

CYSTIC FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR GENE MUTATIONS

QUEST DIAGNOSTICS INVESTM...

1. A method of detecting the presence of a gIVS6a+415_IVS10+2987Dup26817bp mutation in the cystic fibrosis transmembrane regulatory
(CFTR) gene in a human individual comprising:
(a) amplifying a nucleic acid molecule comprising the sequence of SEQ ID NO: 2 from a nucleic acid sample obtained from the
human individual using:

(i) a forward primer that comprises the sequence of SEQ ID NO: 89 and a reverse primer that hybridizes to intron 6b or exon
6b of the CFTR gene; or

(ii) a forward primer that hybridizes to intron 9 or exon 10 of the CFTR gene and a reverse primer that comprises the sequence
of SEQ ID NO: 90; and

(b) detecting the amplified nucleic acid molecule, if present, thereby detecting the gIVS6a+415_IVS10+2987Dup26817bp mutation,
wherein detecting the amplified nucleic acid comprising the sequence of SEQ ID NO: 2 indicates the presence of the gIVS6a+415_IVS10+2987Dup26817bp
mutation.

US Pat. No. 9,177,766

MASS SPECTROMETRIC QUANTITATION ASSAY FOR METABOLITES OF LEFLUNOMIDE

Quest Diagnostics Investm...

1. A method for determining the amount of teriflunomide in a sample by mass spectrometry, said method comprising:
a. subjecting the sample to ionization in positive ion mode under conditions suitable to produce one or more ions detectable
by mass spectrometry;

b. determining the amount of one or more ions by mass spectrometry, wherein said one or more ions determined by mass spectrometry
are selected from the group consisting of ions with mass to charge ratios (m/z) of 271.3 ±0.50, 162.1 ±0.50, and 142.2 ±0.50;
and

c. using the amount of the one or more ions determined in step (b) to determine the amount of teriflunomide in the sample.
US Pat. No. 9,097,732

MASS SPECTROMETRIC DETERMINATION OF EICOSAPENTAENOIC ACID AND DOCOSAHEXAENOIC ACID

Quest Diagnostics Investm...

1. A method for determining the amount of docosahexaenoic acid (DHA) in a human serum or plasma sample by mass spectrometry,
the method comprising:
(i) ionizing DHA from the sample to generate one or more DHA ions detectable by mass spectrometry;
(ii) determining the amount of said one or more DHA ions by single mass spectrometry; and
(iii) relating the amount of DHA ions to the amount of DHA in the sample;wherein said method has a limit of detection for DHA in the sample at a concentration of about 5 ?mol/L or less.

US Pat. No. 9,057,732

MASS SPECTROMETRY ASSAY FOR CONGENITAL ADRENAL HYPERPLASIA

Quest Diagnostics Investm...

1. A method for determining the amount of analytes in a sample by mass spectrometry, the method comprising:
subjecting the sample to an ionization source under conditions suitable to produce one or more ions detectable by mass spectrometry
from each of the analytes, wherein the analytes are not derivatized prior to ionization;

determining by tandem mass spectrometry the amount of the one or more ions from each of the analytes; and
using the determined amount of the one or more ions to determine the amount of each of the analytes in the sample;
wherein the analytes comprise at least two analytes selected from pregnenolone, 17-OH pregnenolone, progesterone, 17-OH progesterone,
dehydroepiandrosterone (DHEA), androstenedione, testosterone, 11-deoxycortisol, deoxycorticosterone, corticosterone, cortisone,
and cortisol.

US Pat. No. 9,244,084

METHODS FOR DETECTING VITAMIN D METABOLITES BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of 25-hydroxyvitamin D3 or 25-hydroxyvitamin D2 or both in a sample by tandem mass spectrometry, comprising:
(a) generating a precursor ion of underivatized 25-hydroxyvitamin D3 or 25-hydroxyvitamin D2 by ionizing said 25-hydroxyvitamin D3 or 25-hydroxyvitamin D2 with atmospheric pressure chemical ionization (APCI), wherein said precursor ion comprises an ion having a mass to charge
ratio of 383.16±0.5 for 25-hydroxyvitamin D3 or 395.30±0.5 for 25-hydroxyvitamin D2;

(b) generating one or more fragment ions of the precursor ion;
(c) detecting the amount of one or more of the ions generated in step (a) or (b) or both and relating the detected ions to
the amount of 25-hydroxyvitamin D3 or 25-hydroxyvitamin D2 or both in the sample;

wherein the method is capable of quantitating 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 at concentrations of less than 4 ng/ml in the sample.

US Pat. No. 9,464,331

DIRECT AMPLIFICATION AND DETECTION OF VIRAL AND BACTERIAL PATHOGENS

QUEST DIAGNOSTICS INVESTM...

1. A method for identifying the presence or absence of a target nucleic acid from a microorganism in a biological sample obtained
from a human, said method comprising:
(a) contacting the sample with a DNA polymerase and a buffer under conditions suitable for amplification of the target nucleic
acid from the sample without extracting the target nucleic acid from the sample;

(b) thermocycling the sample from step (a) such that the target nucleic acid, if present, is amplified; and
(c) detecting the amplified target nucleic acid, if present, produced from step (b),
wherein nucleic acid in the sample is not extracted from the sample prior to amplification, and wherein said buffer comprises
a cationic surfactant and a scorpion primer specific for the target nucleic acid.

US Pat. No. 9,459,267

QUANTITATION OF TAMOXIFEN AND METABOLITES THEREOF BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining an amount of norendoxifen in a human sample by mass spectrometry, comprising:
(a) ionizing said norendoxifen to produce one or more norendoxifen ions detectable by mass spectrometry;
(b) detecting the amount of the norendoxifen ion(s) from step (a) by mass spectrometry; wherein the amount of the ion(s) detected
is related to the amount of norendoxifen in said sample, wherein the method has a limit of quantitation less than or equal
to 5 ng/mL.

US Pat. No. 9,354,200

DETECTION APPARATUS FOR DIFFERENTIAL-CHARGED PARTICLE MOBILITY ANALYZER

QUEST DIAGNOSTICS INVESTM...

1. A method for absolute quantitation of a number of lipoprotein particles in a biological sample, said method comprising:
a) contacting said sample with a first fluorophore labeled entity capable of integrating into a lipoprotein particle of interest
in the sample to generate a first fluorescently labeled lipoprotein particle;

b) introducing the first fluorescently labeled lipoprotein particles to an apparatus for differential-charged particle mobility
analysis and fluorescence detection; and

c) quantitating the number of fluorescently labeled lipoprotein particles in the apparatus;
wherein said lipoprotein particles are Lipoprotein (a) particles; and said fluorophore labeled entity is an aptamer or antibody
capable of specifically binding Apolipoprotein (a).

US Pat. No. 9,234,901

MASS SPECTROMETRY METHOD FOR MEASURING VITAMIN B6 IN BODY FLUIDS

Quest Diagnostics Investm...

1. A method for detecting the presence or amount of pyridoxal 5?-phosphate in a body fluid sample by tandem mass spectrometry,
comprising:
(i) purifying said sample, wherein purifying comprises:
(a) applying a body fluid sample and a first solvent to an extraction column under conditions suitable to reversibly retain
pyridoxal 5?-phosphate on said extraction column;

(b) applying a second solvent to said extraction column under conditions suitable to elute retained pyridoxal 5?-phosphate
from the extraction column, wherein said first and second solvents are different;

(c) applying eluted pyridoxal 5?-phosphate from the extraction column to an analytical column for chromatographic separation;
and

(d) applying a third solvent to said analytical column to elute pyridoxal 5?-phosphate from the analytical column;
(ii) generating a parent ion of said pyridoxal 5?-phosphate from said purified sample;
(iii) generating one or more daughter ions of said parent ion; and
(iv) detecting the presence or amount of one or more said ions generated in step (ii) or step (iii) or both, and relating
the detected ions to the presence or amount of said pyridoxal 5?-phosphate in said sample.

US Pat. No. 9,207,221

METHODS OF DETECTING REVERSE TRIIODOTHYRONINE BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of reverse triiodothyronine (rT3) in a sample by tandem mass spectrometry, said method
comprising:
a. subjecting the rT3 from the sample to liquid chromatography;
b. ionizing rT3 from the sample and an internal standard to generate at least one rT3 ion and at least one internal standard
ion detectable by tandem mass spectrometry;

c. determining the amount of said at least one rT3 ion and the amount of said at least one internal standard ion by tandem
mass spectrometry; and

d. determining the amount of rT3 in the sample, comprising comparing the amount of said at least one rT3 ion and the amount
of said at least one internal standard ion.

US Pat. No. 9,481,914

METHODS FOR THE DIAGNOSIS OF BACTERIAL VAGINOSIS

Quest Diagnostics Investm...

1. A method for identifying bacterial vaginosis in a human female subject, the method comprising:
a. obtaining a vaginal swab sample from the subject;
b. measuring levels of at least two of Atopobium vaginae, Megasphaera genus, or Gardnerella vaginalis, and one or more Lactobacilli species selected from the group consisting of Lactobacillus acidophilus, Lactobacillus crispatus, and Lactobacillus jensenii in the sample, wherein the levels of at least two of Atopobium vaginae, Megasphaera genus, and/or Gardneralla vaginalis, and of one or more Lactobacilli species are detected with one or more oligonucleotides capable of specifically hybridizing to a target nucleic acid sequence
from Atopobium vaginae, Megasphaera genus, Gardneralla vaginalis, Lactobacillus acidophilus, Lactobacillus crispatus, and/or Lactobacillus jensenii, wherein levels of Mobiluncus sp are not measured; wherein the levels are determined with one or more oligonucleotide primers selected from SEQ ID NOs:
1, 2, 4, 5, 11, 12, 14 15, 17, and 18 and complements thereof;

c. calculating a diagnostic score as the ratio of a logarithmic function of the levels of the one or more Lactobacilli species and a logarithmic function of the levels of at least two of Atopobium vaginae, Megasphaera genus, and/or Gardneralla vaginalis; and

d. identifying the subject as having bacterial vaginosis with the diagnostic score lower than a reference score.
US Pat. No. 9,315,802

RNA ISOLATION FROM SOLUBLE URINE FRACTIONS

QUEST DIAGNOSTICS INVESTM...

1. A method for processing urine for detection of RNA expression associated with a benign prostate hyperplasia, comprising:
a) separating urine sediment from the soluble urine fraction of a urine sample obtained from an individual suspected of having
cancer, wherein RNA associated with the benign prostate hyperplasia is present in the soluble urine fraction; and

b) concentrating the soluble urine fraction by ultrafiltration to produce a soluble urine concentrate, wherein the volume
of the soluble urine concentrate is reduced at least 50% from the original urine volume,

wherein the benign prostate hyperplasia-associated RNA is selected from the group consisting of HSPD1, IMPDH2, PDLIM5, and
UAP1.

US Pat. No. 9,194,006

DIRECT AMPLIFICATION AND DETECTION OF VIRAL AND BACTERIAL PATHOGENS

QUEST DIAGNOSTICS INVESTM...

1. A method for identifying the presence or absence of a target nucleic acid from a microorganism in a biological sample obtained
from a human, said method comprising:
(a) contacting the biological sample with a reaction mixture containing a DNA polymerase and a buffer to form a sample mixture
under conditions suitable for amplification of the target nucleic acid from the biological sample without extracting the target
nucleic acid from the biological sample;

(b) thermocycling the sample mixture from step (a) such that the target nucleic acid, if present, is amplified; and
(c) detecting the amplified target nucleic acid, if present, produced from step (b),
wherein the biological sample is selected from the group consisting of whole blood, plasma, serum, and cerebrospinal fluid
(CSF),

wherein the nucleic acid in the biological sample is not extracted from the biological sample prior to amplification;
wherein the biological sample is not diluted prior to contacting the biological sample with the reaction mixture;
wherein 20-30% of the total volume of the sample mixture following step (a) is the biological sample;
wherein the target nucleic acid is not present in a biological sample that does not contain the microorganism; and
wherein the reaction mixture contains a cationic surfactant.
US Pat. No. 9,145,591

DETECTION OF MYCOBACTERIUM TUBERCULOSIS COMPLEX NUCLEIC ACIDS

QUEST DIAGNOSTICS INVESTM...

1. A kit for identifying Mycobacterium tuberculosis complex (MT complex) nucleic acid in a sample, said kit comprising:
(a) a forward oligonucleotide primer that hybridizes to a sequence of at least 15 nucleotides of nucleotide 81-99 of SEQ ID
NO: 5 or the complement of a sequence of at least 15 nucleotides of nucleotides 81-99 of SEQ ID NO: 5, and

(b) a reverse oligonucleotide primer that hybridizes to a sequence of at least 15 nucleotides 165-182 of SEQ ID NO: 5 or the
complement of a sequence of at least 15 nucleotides of nucleotides 165-182,

and further comprising an oligonucleotide probe comprising SEQ ID NO: 3 or the complement of SEQ ID NO: 3, wherein said oligonucleotide
probe is labeled with a donor fluorophore and a quenching moiety.

US Pat. No. 10,072,308

MOLECULAR DETECTION OF ENTEROVIRUS AND PARECHOVIRUS

Quest Diagnostics Investm...

1. A method for determining the presence or absence of an enterovirus and/or a parechovirus in a sample, the method comprising:(a) amplifying enteroviral nucleic acids, if present in the sample, with at least one first pair of primers, wherein the first pair of primers comprises:
(i)a first primer comprising SEQ ID NO:1, and
(ii) a second primer comprising a 5? quencher dye, a first probe sequence flanked by two self-complementary nucleotide sequences of at least four nucleotides in length, a fluorophore and a primer sequence comprising SEQ ID NO:2; and/or
(b) amplifying parechoviral nucleic acids, if present in the sample, with at least one second pair of primers, wherein the second pair of primers comprises:
(i) first primer comprising SEQ ID NO:4, and
(ii) a second primer comprising a 5? quencher dye, a second probe sequence flanked by two self-complementary nucleotide sequences of at least four nucleotides in length, a fluorophore and a primer sequence comprising SEQ ID NO: 5 or the full complement thereof,
wherein detection of amplified enteroviral and/or parechoviral nucleic acids indicates the presence of enterovirus and/or parechovirus, respectively.

US Pat. No. 9,535,077

VITAMIN D METABOLITE DETERMINATION UTILIZING MASS SPECTROMETRY FOLLOWING DERIVATIZATION

Quest Diagnostics Investm...

1. A method for determining an amount of one or more vitamin D metabolites in a sample by mass spectrometry, the method comprising:
(i) subjecting the sample and an internal standard to a Cookson-type derivatizing reagent under conditions sufficient to generate
one or more Cookson-type vitamin D metabolite derivatives and one or more Cookson-type internal standard derivatives, wherein
the internal standard comprises at least one of 25OHD2-[6, 19, 19]-2H3, 25OHD2-[26, 26, 26, 27, 27, 27]-2H6, 25OHD3[6, 19, 19]-2H3, 25OHD3-[26, 26, 26, 27, 27, 27]-2H6, 1?,25(OH)2D3-[6, 19, 19]-2H3, 1?,25(OH)2D2-[26, 26, 26, 27, 27, 27]-2H6, 1?,25(OH)2D3-[6, 19, 19]-2H3 and 1?,25(OH)2D2-[6, 26, 26, 27, 27, 27]-2H6;

(ii) subjecting the one or more Cookson-type vitamin D metabolite derivatives and the one or more Cookson-type internal standard
derivatives to turbulent flow liquid chromatography (TFLC);

(iii) ionizing the one or more Cookson-type vitamin D metabolite derivatives and the one or more Cookson-type internal standard
derivative to generate respectively one or more Cookson-type vitamin D metabolite derivative ions and one or more Cookson-type
internal standard derivative ions detectable by mass spectrometry;

(iv) determining an amount of the one or more of the Cookson-type vitamin D metabolite derivative ions and an amount of the
one or more Cookson-type internal standard derivative ions by mass spectrometry; and

(v) relating the amounts of the Cookson-type vitamin D metabolite derivative ions and the one or more Cookson-type internal
standard derivative ions determined in (iv) to the amount of a vitamin D metabolite in the sample.

US Pat. No. 9,488,656

BCR-ABL TRUNCATION MUTATIONS

QUEST DIAGNOSTICS INVESTM...

1. A method for determining the prognosis of a human patient diagnosed as having a myeloproliferative disease and having a
BCR-ABL gene translocation, comprising:
(a) assaying a nucleic acid sample comprising a BCR-ABL nucleic acid obtained from the patient to determine the presence of
a BCR-ABL Del 2595-2779 deletion mutation, wherein assaying comprises:

(i) contacting the BCR-ABL nucleic acid sample or a BCR-ABL nucleic acid isolated therefrom with a detectably labeled nucleic
acid probe that specifically hybridizes to a mutant BCR-ABL nucleic acid comprising the deletion mutation, if present, but
not to a wild-type BCR-ABL nucleic acid comprising SEQ ID NO: 1, wherein the detectable labeled nucleic acid probe comprises
25 contiguous nucleotides of SEQ ID NO: 4; and

(ii) detecting the BCR-ABL Del 2595-2779 deletion mutation when a hybrid is formed between the detectably labeled nucleic
acid probe and the mutant BCR-ABL; and

(b) identifying the patient as having a poor prognosis when the BCR-ABL Del 2595-2779 deletion mutation is present.

US Pat. No. 9,449,801

MASS SPECTROMETRIC DETERMINATION OF FATTY ACIDS

Quest Diagnostics Investm...

1. A method for determining an amount of one or more very long chain fatty acids (VLCFA) and/or branched chain fatty acids
(BCFA) in a sample by mass spectrometry, the method comprising:
(a) subjecting the sample to an ionization source operating in positive ionization mode to generate one or more VLCFA and/or
BCFA ions detectable by mass spectrometry;

(b) determining the amount of the one or more VLCFA and/or BCFA ions by mass spectrometry; and
(c) relating the amount of the one or more VLCFA and/or BCFA ions determined in step (b) to the amount of the VLCFA and/or
BCFA in the sample;

wherein the VLCFA and/or BCFA are underivatized prior to ionization.
US Pat. No. 9,410,203

CYSTIC FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR GENE MUTATIONS

QUEST DIAGNOSTICS INVESTM...

1. A method of detecting a CFTR mutation, comprising:
(a) contacting a biological sample comprising a CFTR nucleic acid obtained from a human with a detectably labeled nucleic
acid probe that specifically hybridizes to a mutant CFTR nucleic acid comprising a 4177delG deletion mutation, if present,
but not to a wild-type CFTR nucleic acid, wherein the detectably labeled nucleic acid probe comprises a sequence of 8-20 nucleotides
that is fully complementary to a portion of the mutant CFTR nucleic acid and comprises the 4177delG mutation: and

(b) detecting the CFTR 4177delG deletion mutation when a hybrid is formed between the detectably labeled nucleic acid probe
and the mutant CFTR nucleic acid.

US Pat. No. 9,228,230

FLUORESCENCE ENERGY TRANSFER BY COMPETITIVE HYBRIDIZATION

QUEST DIAGNOSTICS INVESTM...

1. A method for real-time monitoring of nucleic acid amplification comprising:
(a) reverse transcribing a viral RNA to DNA;
(b) amplifying a target nucleic acid sequence of the DNA, using a thermostable nucleic acid polymerase, in the presence of
a first oligonucleotide probe and a second oligonucleotide probe, wherein

said first probe;
i) is capable of hybridizing to said target nucleic acid; and
ii) comprises a fluorophore;
said second probe;
i) is capable of hybridizing to said first probe;
ii) has a quencher molecule which quenches said first probe fluorophore when said first and second probes are hybridized to
each other, wherein said fluorophore and said quencher are within fifteen base pairs of each other when said first probe and
said second probe are hybridized to each other; and

iii) is at least two nucleotides shorter in length than said first probe; and
wherein the amplification is carried out with an annealing temperature that is about 5° C. to about 10° C. below the melting
temperature of said probes; and

(c) detecting fluorescence of said first probe fluorophore in real-time to monitor amplification, wherein an increase in fluorescence
correlates with amplification.

US Pat. No. 9,063,119

METHODS FOR DETECTING VITAMIN C BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining an amount of vitamin C in a test sample by mass spectrometry, the method comprising:
(i) ionizing vitamin C in the test sample to generate at least one vitamin C ion detectable by mass spectrometry;
(ii) determining the amount of the at least one vitamin C ion by mass spectrometry; and
(iii) using the amount of vitamin C ion(s) from step (ii) to determine the amount of vitamin C in the test sample.
US Pat. No. 9,583,322

KISSPEPTIN-54 DETECTION BY TANDEM MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the presence of or risk of developing preeclampsia by mass spectrometry, said method comprising:
(a) subjecting a body fluid or tissue sample obtained from a patient to ionization under conditions suitable to produce one
or more multiply charged kisspeptin-54-derived peptide ions detectable by mass spectrometry;

(b) determining by mass spectrometry the amount of one or more multiply charged ions selected from the group of ions with
a charge consisting of 4+, 5+, 6+, and 7+ from each of said one or more kisspeptin-54-derived peptides; and

(c) using the amount of the determined ions to determine the amounts of the corresponding one or more kisspeptin-54-derived
peptides in the sample;
wherein elevated levels of one or more kisspeptin-54-derived peptides in the sample relative to normal levels indicate the
presence of or risk of developing preeclampsia.

US Pat. No. 9,250,211

MAGNETIC SEPARATION OF LIPOPROTEINS USING DEXTRAN SULFATE

QUEST DIAGNOSTICS INVESTM...

1. A method for purifying lipoproteins suitable for differential charged-particle mobility analysis of lipoprotein class and
subclass, said method comprising:
(a) adding one or more cations and dextran sulfate to a sample comprising lipoproteins to form a precipitation mixture;
(b) contacting the precipitation mixture with a microbead under conditions for one or more classes or subclasses of lipoproteins
in said mixture to bind to an inert outer layer of the microbead, wherein said inert outer layer comprises carboxylate or
NH2, but does not include a lipoprotein binding lipoprotein-capture ligand;

(c) separating lipoproteins bound to the microbead from said mixture to obtain a microbead with bound lipoproteins; and
(d) removing said bound lipoproteins from the microbead;
wherein said lipoproteins bound to the microbead comprise HDL and one or more selected from the group consisting of LDL, Lp(a),
IDL and VLDL.

US Pat. No. 9,506,937

MASS SPECTROMETRY OF STEROIDAL COMPOUNDS IN MULTIPLEXED PATIENT SAMPLES

Quest Diagnostics Investm...

1. A method for determining the amount of a steroidal compound in each of a plurality of human samples with a single mass
spectrometric assay, the method comprising:
i) subjecting each of a plurality of human samples to a different Cookson-type derivatizing agent to generate a differently
derivatized steroidal compound in each of the plurality of samples;

ii) combining the plurality of samples to form a multiplex sample; and
iii) quantifying the amount of the steroidal compound in each sample by mass spectrometry.

US Pat. No. 9,382,586

METHOD TO DETECT REPEAT SEQUENCE MOTIFS IN NUCLEIC ACID

QUEST DIAGNOSTICS INVESTM...

19. A method for determining the presence or absence of an expansion of the CGG repeat tract in the 5?-untranslated region
(5?-UTR) of fragile X related mental retardation 1 (FMRI) gene in an individual, said method comprising:
a) amplifying all or a portion of said 5?-UTR CGG repeat tract in the presence of a deoxy GTP analog, using as template genomic
DNA comprising the FMR1 gene obtained from a biological sample from said individual, said amplifying being achieved with at
least a primer pair, said primer pair comprising an upstream primer and a downstream primer, wherein:

(i) the downstream primer comprises a CCG repeat segment that comprises at least two CCG triplets and less than four consecutive
CCG triplets, and hybridizes to the junction between the 3? end of the CGG repeat tract in the FMR1 gene and sequence directly
3? thereto, wherein said CCG repeat segment in said downstream primer comprises a nucleotide sequence of any one of SEQ ID
NOs. 79-94; or

(ii) the upstream primer comprises a CGG repeat segment that comprises at least two CGG triplets and less than four consecutive
CGG triplets, and hybridizes to the junction between the 5? end of the CGG repeat tract in the FMR1 gene and sequence directly
5? thereto, wherein said CGG repeat segment in said upstream primer comprises a nucleotide sequence of any one of SEQ ID NOs.
97-112;

said amplifying generates amplicons that contain a portion of the CGG repeat tract;
b) separating said amplicons according to size by capillary electrophoresis; and
c) detecting the presence or absence of an expansion of the CGG repeat tract, wherein the size of said amplicons indicates
the presence or absence of expansion of said CGG repeat tract in said 5?-UTR of FMRI gene.

US Pat. No. 9,255,926

HEMATOPOIETIC CELL PHENOTYPING USING CIRCULATING CELL-FREE MARKERS

QUEST DIAGNOSTICS INVESTM...

1. A method for predicting survival or remission duration in a patient with myelodysplastic syndrome (MDS), the method comprising:
(a) assaying a blood plasma sample from a patient diagnosed as having MDS to determine the level of circulating cell-free
CD4 marker in said sample, wherein assaying comprises contacting the blood plasma sample with an antibody specific for CD4,

(b) correlating the level of circulating cell-free CD4 marker determined in step (a) with the patient's predicted survival
or remission duration, and

(c) identifying the patient as
(i) having a longer predicted survival or remission duration if the level of cell-free CD4 is determined to be greater than
1029 U/?l, whereby a level greater than 1029 U/?l indicates that the patient will have a longer predicted survival or remission
duration than is indicated by a level less than 1029 U/?l; or

(ii) having a shorter predicted survival or remission duration if the level of cell-free CD4 is determined to be less than
1029 U/?l, whereby a level less than 1029 U/?l indicates that the patient will have a shorter survival or remission duration
than is indicated by a level greater than 1029 U/?l.

US Pat. No. 9,696,325

MASS SPECTROMETRIC DETERMINATION OF EICOSAPENTAENOIC ACID AND DOCOSAHEXAENOIC ACID

Quest Diagnostics Investm...

1. A method for determining the amount of docosahexaenoic acid (DHA) in a human serum or plasma sample by mass spectrometry,
the method comprising:
(i) ionizing DHA from the sample to generate one or more DHA ions detectable by mass spectrometry;
(ii) determining the amount of said one or more DHA ions by single mass spectrometry, wherein said one or more ions comprises
an ion with a mass to charge ratio (m/z) of 327.2±0.5; and

(iii) relating the amount of DHA ions to the amount of DHA in the sample, wherein said method has a limit of detection for
DHA in the sample of about 5 ?mol/L or less.

US Pat. No. 9,580,740

THYROGLOBULIN QUANTITATION BY MASS SPECTROSCOPY

Quest Diagnostics Investm...

1. A method of determining the amount of thyroglobulin in a test sample, comprising:
(a) digesting thyroglobulin (Tg) in the test sample;
(b) enriching a digested Tg peptides comprising an amino acid sequence VIFDANAPVAVR (SEQ ID NO:1);
(c) ionizing the Tg peptide and an isotopically labeled internal standard, wherein the internal standard comprises a synthetic
winged peptide comprising an amino acid sequence VIFDANAPVAVR (SEQ ID NO:1) that is digested with thyroglobulin from the test
sample; and

(d) quantifying the amount of the Tg peptide ion from step (c) by mass spectrometry; wherein the amount of the Tg peptide
ion detected in step (d) is related to the amount of thyroglobulin in said test sample.

US Pat. No. 9,447,471

MICRORNA PROFILING FOR DIAGNOSIS OF DYSPLASTIC NEVI AND MELANOMA

QUEST DIAGNOSTICS INVESTM...

1. A method for distinguishing melanoma cells from dysplastic nevi cells in a skin lesion sample from a subject comprising:
(a) performing reverse transcriptase polymerase chain reaction (RT-PCR) to amplify nucleic acids from a skin sample isolated
from a skin lesion, wherein the skin sample is previously identified as containing melanoma cells and/or dysplastic nevi cells;

(b) assaying the expression level of two or more miRNAs in the skin sample by detecting the amplified nucleic acids of (a),
wherein the two or more miRNAs are selected from the group consisting of:

i. miR-150;
ii. miR-149-star;
iii. miR-1308;
iv. miR-191;
v. miR-1228-star;
vi. ENSG00000199411_s;
vii. miR-1268;
viii. miR-923;
ix. miR-23a;
x. miR-132;
xi. miR-1207.5p;
xii. miR-342.3p;
xiii. U38B; and
xiv. miR-155,
wherein assaying the level of expression comprises using a detectably labeled probe specific for each miRNA assayed;
(c) quantitatively comparing the levels of the two or more miRNAs in the skin lesion sample as assayed in (b) to the levels
of each corresponding miRNA in a normal skin or nevus reference sample; and

(d) identifying the skin sample as containing:
(i) melanoma cells if the levels of each of the two or more miRNAs in the skin sample differs from the normal skin or nevus
reference sample, wherein miR-150, miR-191, miR-23a, miR-132, miR-342.3p, and miR-155 are increased and miR-149-star, miR-1308,
miR-1228-star, ENSG00000199411_s, miR-1268, miR-923, miR-1207.5p, and U38B are decreased relative to the normal skin or nevus
reference sample; or

(ii) dysplastic nevi cells and/or no melanoma cells if the levels of each of the two or more miRNAs in the skin sample do
not differ from the normal skin or nevus reference sample.

US Pat. No. 9,175,350

EML4-ALK TRANSLOCATIONS IN LUNG CANCER

QUEST DIAGNOSTICS INVESTM...

1. A method for diagnosing a non-small cell lung cancer or susceptibility to non-small cell lung cancer in human subject comprising:
(a) performing a nucleic acid detection assay on a nucleic acid sample from a human subject to detect the presence of an EML4-ALK
gene fusion in the nucleic acid sample, wherein the EML4-ALK gene fusion is (i) an E17;ins30A20 gene fusion between exon 17
of EML4 and intron 19 of ALK having a breakpoint region comprising SEQ ID NO: 1, or (ii) an E17ins30;ins65A20 gene fusion
between exon 17 of EML4 and intron 19 of ALK having a breakpoint region comprising SEQ ID NO:2; and

(b) diagnosing the subject as having or being susceptible to non-small cell lung cancer based on the presence of the EML4-ALK
gene fusion in the nucleic acid sample,

wherein the nucleic acid detection assay comprises amplification of a nucleic acid molecule with at least a primer pair, said
primer pair comprising a forward primer comprising the nucleotide sequence set forth in SEQ ID NO: 19 and a reverse primer
that hybridizes to exon 20 of ALK to produce amplified nucleic acid, and wherein the amplified nucleic acid comprises the
sequence of SEQ ID NO: 1 or 2.

US Pat. No. 9,733,249

MPL MUTATIONS IN JAK2 V617F NEGATIVE PATIENTS WITH MYELOPROLIFERATIVE DISEASE

Quest Diagnostics Investm...

1. A method of detecting a T1588 T1599 del/ins6 mutation in the myeloproliferative leukemia (MPL) gene in an individual, comprising:
(a) contacting a sample containing a MPL nucleic acid from an individual suspected of having a mutation in the MPL gene with
a detectably labeled nucleic acid probe that hybridizes to a mutant MPL nucleic acid comprising the T1588 T1599 del/ins6 mutation
but not to a wild type MPL nucleic acid, wherein the nucleic acid probe comprises the sequence set forth in SEQ ID NO:10;
and

(b) detecting a hybrid formed between the detectably labeled nucleic acid probe and the mutant MPL nucleic acid.

US Pat. No. 9,638,705

PROGNOSTIC ASSAYS FOR MAINTENANCE HEMODIALYSIS PATIENTS

QUEST DIAGNOSTICS INVESTM...

1. A method for predicting mortality risk in a maintenance hemodialysis (MHD) patient comprising:
a. separating lipoprotein particles from non-lipoprotein particles in a plasma sample from an MHD patient;
b. irradiating with alpha radiation and fractionating the lipoprotein particles according to particle diameter using on mobility
lipoprotein fractionation;

c. determining a very small low density lipoprotein (vs-LDL) particle concentration in the plasma sample by detecting and
counting the fractionated lipoprotein particles having a particle diameter of 180.0-208.2 A;

d. determining a large low density lipoprotein (1-LDL) particle concentration in the plasma sample by detecting and counting
the fractionated lipoprotein particles having a particle diameter of 220.0-233.3 A; and

e. identifying the MHD patient as having:
i. increased risk of mortality when the sample vs-LDL particle concentration is greater than a reference vs-LDL particle concentration;
ii. reduced risk of mortality when the sample 1-LDL particle concentration is greater than a reference 1-LDL particle concentration;
or

iii. no change in the risk of mortality when the sample vs-LDL particle concentration is less than or equal to the reference
vs-LDL particle concentration and the sample 1-LDL particle concentration is less than the reference 1-LDL concentration,

whereby a vs-LDL particle concentration in a plasma sample from a MHD patient that is greater than the reference vs-LDL particle
concentration and a 1-LDL

particle concentration in a plasma sample from a MHD patient that is greater than the reference 1-LDL particle concentration
indicates an reduced risk of mortality in the MHD patient.

US Pat. No. 9,593,375

NUCLEIC ACID ANALYSIS USING EMULSION PCR

QUEST DIAGNOSTICS INVESTM...

1. A method of nucleic acid analysis comprising:
(a) creating an emulsion from an aqueous nucleic acid sample comprising at least one chromosome or a fragment thereof greater
than 10 kilobase pairs in length and containing a target region, wherein each emulsion droplet contains an average of between
0-2 nucleic acids from the aqueous nucleic acid sample and wherein the at least one chromosome or a fragment thereof is contained
within an emulsion droplet of the emulsion;

(b) amplifying the target region within the emulsion;
(c) quantifying the number of emulsion droplets that comprise an amplified target region; and
(d) calculating the ratio of droplets containing amplified nucleic acid from the target region to total droplets.
US Pat. No. 10,093,980

BCR-ABL TRUNCATION MUTATIONS

Quest Diagnostics Investm...

1. A method for determining the prognosis of a human patient diagnosed as having a myeloproliferative disease and having a BCR-ABL gene translocation, comprising:(a) assaying a nucleic acid sample comprising a BCR-ABL nucleic acid obtained from the patient to determine the presence of one or more BCR-ABL truncation mutations selected from the group consisting of 2417insCAGG, Del 2596-2597, and C2506T, wherein assaying comprises:
(i) performing a nucleic acid amplification reaction on a BCR-ABL mRNA sample, or cDNA derived therefrom, with at least one primer pair, comprising (1) a forward primer comprising SEQ ID NO: 23 and a reverse primer that binds to the junction of ABL exon 9 and exon 10 or (2) a forward primer that binds to ABL exon 4 and a reverse primer comprising SEQ ID NO: 22;
(ii) contacting the amplification product of (i) with a detectably labeled nucleic acid probe that specifically hybridizes to a mutant BCR-ABL nucleic acid comprising the mutation, if present, but not to a wild-type BCR-ABL nucleic acid comprising SEQ ID NO: 1; and
(iii) detecting the BCR-ABL truncation mutation when a hybrid is formed between the detectably labeled nucleic acid probe and the amplification product of (i); and
(b) identifying the patient as having a poor prognosis when the BCR-ABL truncation mutation is present.
US Pat. No. 10,030,276

DETECTION OF MYCOBACTERIUM TUBERCULOSIS COMPLEX NUCLEIC ACIDS

Quest Diagnostics Investm...

1. A solution for amplification of a Mycobacterium tuberculosis complex (MT complex) nucleic acid in a sample, comprising:(a) a forward oligonucleotide primer that hybridizes to a sequence of at least 15 nucleotides of nucleotides 81-99 of SEQ ID NO:5 or the complement of a sequence of at least 15 nucleotides of nucleotides 81-99 of SEQ ID NO:5, and
(b) a reverse oligonucleotide primer that that hybridizes to a sequence of at least 15 nucleotides of nucleotides 165-182 of SEQ ID NO:5 or the complement of a sequence of at least 15 nucleotides of nucleotides 165-182; and
(c) an oligonucleotide probe that detects an amplicon produced by amplification with the forward oligonucleotide primer and the reverse oligonucleotide primer, wherein the oligonucleotide probe is labeled with a reporter dye.
US Pat. No. 9,874,569

KISSPEPTIN-54 DETECTION BY TANDEM MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining by mass spectrometry the amount in a sample of one or more kisspeptin-54-derived peptides, said
method comprising:
(a) enriching the concentration of the one or more kisspeptin-54-derived peptides in a sample with an antibody specific for
the N-terminal portion of kisspeptin-54

(b) ionizing the enriched sample to produce one or more multiply charged kisspeptin-54-derived peptide ions detectable by
mass spectrometry;

(c) determining the amount of one or more multiply charged ions selected from the group of ions with a charge consisting of
4+,5+,6+, and 7+ by mass spectrometry, wherein the amount of the ions is used to determine the amounts of the corresponding
one or more kisspeptin-54-derived peptides in the sample.

US Pat. No. 9,863,000

CYSTIC FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR GENE MUTATIONS

Quest Diagnostics Investm...

1. A method for detecting the presence of a gIVS6a+415_IVS10+2987Dup26817 bp mutation in the cystic fibrosis transmembrane
regulatory (CFTR) gene in a human individual comprising:
(a) contacting a nucleic acid sample comprising a CFTR nucleic acid with a detectably labeled nucleic acid probe that hybridizes
to a mutant CFTR nucleic acid comprising the gIVS6a+415_IVS10+2987Dup26817 bp mutation, if present, but not to a wild-type
CFTR nucleic acid, wherein the detectably labeled nucleic acid probe comprises SEQ ID NO: 2; and

(b) detecting the gIVS6a+415_IVS10+2987Dup26817 bp mutation when a hybrid is formed between the detectably labeled nucleic
acid probe and the mutant CFTR nucleic acid.

US Pat. No. 9,541,562

MASS SPECTROMETRY ASSAY FOR CONGENITAL ADRENAL HYPERPLASIA

Quest Diagnostics Investm...

1. A method for determining the amount of a panel of analytes in a sample by mass spectrometry, the method comprising:
ionizing the sample under conditions suitable to produce one or more ions detectable by mass spectrometry from each of the
analytes;

determining by tandem mass spectrometry the amount of the one or more ions from each of the analytes; and
using the determined amount of the one or more ions to determine the amount of each of the analytes in the sample;
wherein the analytes comprise 17-OH progesterone and androstenedione.

US Pat. No. 9,791,464

LIPOPROTEIN ANALYSIS BY DIFFERENTIAL CHARGED-PARTICLE MOBILITY

QUEST DIAGNOSTICS INVESTM...

1. A method for obtaining purified Lp(a), said method comprising:
(a) preparing a centrifuge tube containing a first solution underneath a sample, said sample comprising one or more lipoproteins
and non-lipoprotein components, said first solution having a first density greater than 1.00 g/mL and less than or equal to
about 1.21 g/mL;

(b) subjecting said tube to centrifugation sufficient to cause said non-lipoprotein components to migrate toward the bottom
of the tube and away from said lipoproteins, and collecting the lipoproteins;

(c) forming a precipitate of Lp(a) by admixing the collected lipoproteins with a precipitant for Apo B-containing lipoprotein
under conditions sufficient to cause precipitation of Lp(a);

(d) isolating said Lp(a) containing precipitate from said sample:
(e) solubilizing said Lp(a) containing precipitate;
(f) admixing said solubilized Lp(a) with a solid-support reagent containing a lectin attached to a solid support to allow
formation of a Lp(a)-lectin complex;

(g) isolating said Lp(a)-lectin complex; and
(h) releasing said Lp(a) from said Lp(a)-lectin complex, thereby providing purified Lp(a);
wherein said lectin is selected from the group consisting of wheat germ agglutinin (WGA), lima bean agglutinin (LGA), phytohemagglutinin
(PHA), and horseshoe crab lectin (HCL); and

wherein said releasing comprises disulfide reduction of said Lp(a)-lectin complex.
US Pat. No. 9,783,854

NUCLEIC ACID DETECTION COMBINING AMPLIFICATION WITH FRAGMENTATION

Quest Diagnostics Investm...

35. A method for detecting the presence or absence of a target nucleotide sequence in a sample potentially comprising both
target and non-target nucleotide sequences, wherein the non-target nucleotide sequence comprises a wild-type nucleotide sequence,
and the target nucleic acid comprises a mutant version of the wild-type nucleotide sequence lacking at least one fragmentation
site present in the wild-type nucleotide sequence, and wherein the sample comprises only unamplified nucleic acids, comprising:
a) fragmenting the nucleic acids in the sample under conditions such that fragmentation occurs at the at least one fragmentation
site present in the non-target nucleotide sequence but absent from the target nucleotide sequence;

b) subsequent to step a), combining the product of a) with a polymerase chain reaction (PCR) mixture comprising a pair of
primers specific for the target nucleotide sequence, wherein one primer spans or overlaps the at least one fragmentation site
present in the non-target nucleotide sequence but absent from the target nucleotide sequence, and wherein fragmentation of
the non-target nucleotide sequence in part a) prevents hybridization of said one primer to the non-target nucleotide sequence;

c) subsequent to step b), performing an amplification reaction on the PCR mixture of b); and
d) detecting presence or absence of an amplification product in c), wherein the presence of an amplification product in d)
indicates the presence of the target nucleotide sequence in the sample.

US Pat. No. 9,749,394

AUTOMATED DELIVERY OF ALERTS WITH CONFIRMATION OF RECEIPT

QUEST DIAGNOSTICS INVESTM...

1. A method of transmitting an alert from a computer system that comprises at least one processor, at least one network interface
operatively coupled to at least one of the processors, and a computer-readable storage medium operatively coupled to at least
one of the processors, the method comprising:
transmitting through at least one of the network interfaces, to a client that is a computer system, first information that
comprises an alert;

receiving through at least one of the network interfaces second information indicating that the client has executed instructions
to cause the client to present the alert visually, on an electronic display directly coupled to the client, at a first time,
wherein the second information includes the first time and does not indicate whether the client has received user input acknowledging
receipt of the alert;

in response to receiving the second information, storing in the computer-readable storage medium a record that the first information
was presented, the record comprising the first time;

waiting a predetermined time immediately following transmission of the first information, during which no information is received
through any of the network interfaces indicating that the client has received user input acknowledging receipt of the alert;
and

transmitting through at least one of the network interfaces, to a destination that is not the client, third information indicating
that the alert has not been acknowledged within the predetermined time.

US Pat. No. 9,909,183

CYSTIC FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR GENE MUTATIONS

Quest Diagnostics Investm...

1. A method of detecting a cystic fibrosis transmembrane regulatory (CFTR) mutation, comprising:
(a) contacting a biological sample comprising a CFTR nucleic acid obtained from a human with a detectably labeled nucleic
acid probe that specifically hybridizes to a CFTR nucleic acid comprising a 1342-2A>G mutation, wherein the detectably labeled
nucleic acid probe comprises a sequence of 8-20 nucleotides that is fully complementary to a portion of the CFTR nucleic acid
that comprises the mutation; and

(b) detecting the 1342-2A>G mutation when a hybrid is formed between the probe and the CFTR nucleic acid comprising the 1342-2A>G
mutation.

US Pat. No. 9,797,018

KITS FOR DETECTING MYCOBACTERIUM AVIUM/INTRACELLULARE NUCLEIC ACID

Quest Diagnostics Investm...

1. A kit comprising:
a first oligonucleotide primer that is 20-100 nucleotides in length and comprises SEQ ID NO:4 or the full complement thereof
and a second oligonucleotide primer that is 21-100 nucleotides in length and comprises SEQ ID NO:5 or the full complement
thereof, that, together, constitute a primer pair capable of amplifying wherein both oligonucleotide primers hybridize to
a first target nucleic acid that is the insertion sequence transposase (IS1245) of M. avium or a region thereof;

a third oligonucleotide primer that is 15-100 nucleotides in length and a fourth oligonucleotide primer that is 15-100 nucleotides
in length, that, together constitute a primer pair capable of amplifying wherein both oligonucleotide primers hybridize to
a second target nucleic acid that is the DT1 gene of M. intracellulare, M. avium serovar 2 and M. avium serovar 3 or a region thereof and wherein each primer is at least 75% identical to the corresponding region of the DT1 gene
with which it aligns; and

a fifth oligonucleotide that is 15-70 nucleotides in length, is at least 75% identical to a corresponding region of IS1245,
and hybridizes to an IS1245 amplicon produced with the first and second oligonucleotide primers, wherein the fifth oligonucleotide
comprises one or more detectable labels selected from the group consisting of fluorescent dyes, 32P, 35S, 3H, 14C, 125I, 131I,
electron-dense reagents, enzymes, colorimetric labels, magnetic labels, biotin, dioxigenin, haptens, proteins for which antisera
or monoclonal antibodies are available, and ligands capable of forming a complex with a corresponding receptor; and wherein
the components of the kit are used to detect Mycobacterium avium complex.

US Pat. No. 9,685,311

METHODS FOR DETECTING REVERSE TRIIODOTHYRONINE BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of reverse triiodothyronine (rT3) in a sample by tandem mass spectrometry, said method
comprising:
a. adding an internal standard to the sample;
b. subjecting the sample to protein precipitation;
c. ionizing rT3 and the internal standard to generate at least one rT3 ion and at least one internal standard ion;
d. determining the amount of said at least one rT3 ion and the amount of said at least one internal standard ion by tandem
mass spectrometry; wherein the amount of rT3 in the sample is determined from the amount of said at least one rT3 ion and
the amount of said at least one internal standard ion.

US Pat. No. 9,645,158

METHODS FOR DETECTING VITAMIN C BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining an amount of vitamin C in a sample by tandem mass spectrometry (MS/MS), the method comprising:
(i) purifying vitamin C in the sample;
(ii) ionizing vitamin C to generate at least one vitamin C ion detectable by mass spectrometry;
(iii) determining the amount of the at least one vitamin C ion by tandem mass spectrometry;
wherein the amount of vitamin C in the sample is determined from the amount of vitamin C ion(s) from step (ii).

US Pat. No. 9,619,622

PRINTING FROM A HANDHELD DEVICE VIA A REMOTE SERVER

QUEST DIAGNOSTICS INVESTM...

1. A method of causing printing of a document in response to a request from a handheld device, the method being performed
by a networked computer system that comprises one or more processors, one or more interfaces operatively coupled to at least
one of the processors, and one or more databases operatively coupled to at least one of the processors, the method comprising:
receiving from the handheld device, through one or more of the interfaces, a first one or more queries comprising information
that identifies a document and location information that specifies the location of the handheld device;

retrieving from at least one of the databases information describing a plurality of printing systems;
based on the retrieved information and the location information, identifying a plurality of candidate printing systems;
transmitting to the handheld device, through one or more of the interfaces, information related to the candidate printing
systems;

receiving from the handheld device, through one or more of the interfaces, a second one or more queries comprising information
indicating selection of one of the candidate printing systems; and

transmitting to the selected printing system, through one of the interfaces, information sufficient to cause the selected
printing system to print the document;

wherein the computer system does not receive the document from the handheld device.

US Pat. No. 10,138,519

UNIVERSAL SANGER SEQUENCING FROM NEXT-GEN SEQUENCING AMPLICONS

Quest Diagnostics Investm...

1. A composition comprising:a first oligonucleotide that comprises, in 5? to 3? order,
(a) a first region suitable for use as a Sanger sequencing primer;
(b) a spacer that is between about 5 and about 15 nucleotides long; and
(c) a third region suitable for use as a next generation sequencing (NGS) primer, and
a second oligonucleotide that comprises, in 5? to 3? order,
(a) a first region that is substantially identical to the third region of the first oligonucleotide; and
(b) a second region that is suitable for use as a polymerase chain reaction (PCR) primer to amplify a target nucleotide sequence,
wherein the third region of the first oligonucleotide and the first region of the second oligonucleotide have a melting temperature (Tm) that is at least about 5° C. higher than the Tm of the second region of the second oligonucleotide,
wherein the total length of the spacer and the first and second regions of the second oligonucleotide is at least about 45 nucleotides (nt) long,
wherein the third region of the first oligonucleotide and the first region of the second oligonucleotide are selected from Table 2, and
wherein the third region of the first oligonucleotide and the first region of the second oligonucleotide do not include any sequence that is specific to the target nucleotide sequence.

US Pat. No. 9,921,231

MASS SPECTROMETRY ASSAY FOR CONGENITAL ADRENAL HYPERPLASIA

Quest Diagnostics Investm...

1. A method for determining the amount of a panel of analytes in a sample by mass spectrometry, the method comprising:
(i) purifying the sample by liquid chromatography;
(ii) ionizing the sample under conditions suitable to produce one or more ions detectable by mass spectrometry from each of
the analytes comprising 17-OH progesterone, cortisol, and androstenedione;

(iii) determining by tandem mass spectrometry the amount of the one or more ions from each of the analytes; and
(iv) using the determined amount of the one or more ions to determine the amount of each of the analytes in the sample.
US Pat. No. 9,840,740

CYSTIC FIBROSIS GENE MUTATIONS

QUEST DIAGNOSTICS INVESTM...

1. A method of detecting a mutant cystic fibrosis transmembrane (CFTR) nucleic acid in an individual, comprising: (a) contacting
a biological sample comprising a CFTR nucleic acid from an individual with a detectably labeled nucleic acid probe that specifically
hybridizes to a mutant CFTR nucleic acid comprising the mutation but not to a wildtype CFTR nucleic acid; and the probe comprises
the substitution mutation; and (b) detecting the CFTR substitution mutation in the individual when a hybrid is formed between
the detectably labeled nucleic acid probe and the mutant CFTR nucleic acid, wherein the mutation is c.3017C>A.

US Pat. No. 9,840,741

CIRCULATING MICRORNA AS A MARKER FOR HEPATOCELLULAR CARCINOMA

QUEST DIAGNOSTICS INVESTM...

1. A detection method comprising:
(a) detecting a level of miR-16 miRNA in a serum sample from a human subject diagnosed as having liver cirrhosis by
(i) performing real-time reverse transcription polymerase chain reaction (RT-PCR) on the serum sample to measure a Ct value
for miR-16 miRNA and a control Ct value for U6 small RNA, wherein the Ct value is measured using a detectably labeled probe
specific for each miRNA assayed; and

(ii) calculating a delta Ct value by subtracting the U6 control Ct value from the miR-16 Ct value; and
(b) detecting at least one of the level of ?-fetoprotein (AFP), the level of des-gamma-carboxyprothrombin (DCP), and the percentage
of AFP-L3 in the serum sample by

(i) contacting the serum sample with an antibody that binds to AFP, an antibody that binds to DCP and/or an antibody that
binds to AFP-L3; and

(ii) detecting the binding between AFP and the antibody that binds to AFP, the binding between DCP and the antibody that binds
to DCP and/or the binding between AFP-L3 and the antibody that binds to AFP-L3.

US Pat. No. 9,835,606

QUANTITATION OF TAMOXIFEN AND METABOLITES THEREOF BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of norendoxifen in a human sample by mass spectrometry, comprising:
(a) purifying the sample by liquid chromatography;
(b) ionizing said norendoxifen to produce one or more norendoxifen ions detectable by mass spectrometry;
(c) detecting an amount of the norendoxifen ion(s) from step (b) by mass spectrometry; wherein the amount of the ion(s) detected
is related to the amount of norendoxifen in said sample,

wherein the method has a limit of quantitation of less than or equal to 5 ng/mL.
US Pat. No. 9,702,877

BCR-ABL VARIANTS

QUEST DIAGNOSTICS INVESTM...

1. A method for detecting a 35 INS BCR-ABL splice variant comprising SEQ ID NO:1, comprising:
(a) performing a nucleic acid amplification reaction on a bcr-abl mRNA sample, or cDNA derived therefrom with at least one
primer pair, comprising:

(i) a forward primer that hybridizes to exon 8 of the abl gene and a reverse primer comprising SEQ ID NO: 23; or
(ii) a forward primer comprising SEQ ID NO: 22 that hybridizes to exon 9 of the abl gene, wherein one or both primers of the
primer pair is labeled;

(b) detecting an amplification product that comprises SEQ ID NO: 1, thereby detecting the bcr-abl splice variant.
US Pat. No. 9,663,781

DIAGNOSIS OF PROSTATE CANCER

QUEST DIAGNOSTICS INVESTM...

1. A method for diagnosing prostate cancer in an individual, said method comprising:
(a) separating urine sediment from a soluble urine fraction of a urine sample obtained from an individual suspected of having
prostate cancer;

(b) concentrating the soluble urine fraction by ultrafiltration to produce a soluble urine concentrate, wherein the volume
of soluble urine concentrate is reduced at least 50% from the original urine volume,

(c) detecting the amount of RNA from each of heat shock 60 kDa protein 1 (HSPD1), inosine monophosphate dehydrogenase 2 (IMPDH2),
PDZ and LIM domain 5 (PDLIM5), and UDP-N-acetylglucosamine pyrophosphorylase 1 (UAP1) in the concentrated soluble urine fraction;

(d) comparing the amount of each said RNA to a reference value for each said RNA, wherein said reference value is derived
from the amount of each RNA in a corresponding sample from one or more individuals that do not have prostate cancer; and

(e) diagnosing said individual as having prostate cancer when the amount of said RNA is greater than said reference value
for each RNA.

US Pat. No. 9,617,579

DDR2 MUTATIONS AS TARGETABLE FEATURES OF MELANOMA OR BASAL CELL CARCINOMA

QUEST DIAGNOSTICS INVESTM...

1. A method of detecting a DDR2 mutation in a human suspected of having melanoma, comprising sequencing a nucleic acid encoding
DDR2 protein in a sample from the human, wherein the nucleic acid encodes DDR2 protein that differs from SEQ ID NO:170 by
a mutation selected from the group consisting of R105C, P321L, R458H, S467F, P476S, I488S, F574C, S667F, S674F, R680L, L701F,
R742Q, and T836A.
US Pat. No. 9,914,973

DETECTION OF GENE FUSIONS BY INTRAGENIC DIFFERENTIAL EXPRESSION (IDE) USING AVERAGE CYCLE THRESHOLDS

Quest Diagnostics Investm...

1. A method for detecting a dysregulation in a target gene comprising:
(a) amplifying portions of a 5? region of a transcript of the target gene or a cDNA derived therefrom, if present in a test
sample, with two or more different 5? target primer pairs that are directed to the portions of the 5? region of the target
gene,

(b) amplifying portions of a 3? region of the transcript of the target gene or a cDNA derived therefrom, if present in the
test sample, with two or more different 3? target primer pairs that are directed to the portions of the 3? region of the target
gene,

(c) detecting the amplification products produced by the two or more 5? target primer pairs and the two or more 3? target
primer pairs,

(d) determining the average cycle threshold (Ct) among the two or more 5? target primer pairs and the average Ct among the
two or more 3? target primer pairs, and

(e) identifying the nucleic acid sample as having a gene dysregulation when the average Ct among the two or more 5? target
primer pairs and the average Ct among the two or more 3? target primer pairs as determined in step (d) indicate that the target
gene is dysregulated.

US Pat. No. 9,529,004

METHODS FOR DETECTING VITAMIN D METABOLITES BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of a vitamin D metabolite in a sample by tandem mass spectrometry, comprising:
(a) ionizing the vitamin D metabolite to generate a precursor ion;
(b) generating one or more fragment ions from the precursor ion having a mass-to-charge ratio of 251.30±0.5, 211.35±0.5, 209.20±0.5,
or 179.10±0.5;

(c) detecting the amount of one or more of the ions generated in step (b) and relating the detected ions to the amount of
the vitamin D metabolite in the sample.

US Pat. No. 10,093,985

METHODS FOR DETECTING GENE DYSREGULATION BY INTRAGENIC DIFFERENTIAL EXPRESSION

Quest Diagnostics Investm...

1. A kit comprising:(a) a 5? target primer pair that is complementary to a 5? region of a TMPRSS2 gene transcript, wherein a first primer of the 5? target primer pair comprises the sequence of SEQ ID NO: 1 and a second primer of the 5? target primer pair comprises the sequence of SEQ ID NO: 2;
(b) an oligonucleotide probe that is complementary to an amplification product generated using the 5? target primer pair, and labeled with a donor fluorophore and a quenching moiety;
(c) a 3? target primer pair that is complementary to a 3? region of the TMPRSS2 gene transcript, wherein a first primer of the 3? target primer pair comprises the sequence of SEQ ID NO: 5 and a second primer of the 3? target primer pair comprises the sequence of SEQ ID NO: 6; and
(d) an oligonucleotide probe that is complementary to an amplification product generated using the 3? target primer pair, and labeled with a donor fluorophore and a quenching moiety.

US Pat. No. 9,966,153

GRAPHICAL PRESENTATION OF MEDICAL DATA

Quest Diagnostics Investm...

1. A computerized method of presenting information graphically, comprising:presenting on an electronic display device a flowsheet, comprising
a plurality of identifiers, each of which textually identifies exactly one of at least a plurality of stored series of one or more related data points, wherein each data point comprises a value and temporal information, the temporal information comprising a time, a date, or a time and a date, each of the identifiers being displayed in association with one or more controls, the associated controls together permitting a user to choose between associating the series with a left axis, associating the series with a right axis, and associating the series with no axis;
a plurality of temporal labels, each temporal label indicating at least a time, a date, or a time and a date, the temporal labels together spanning a period of time;
a plurality of numeric values, each numeric value representing one or more data points from a respective exactly one of the plurality of stored series, each numeric value being displayed in visual association with the identifier that identifies the respective series from which the data points were taken, each numeric value further being displayed in visual association with exactly one of the temporal labels, no more than one of the numeric values that are associated with a respective identifier being associated with any respective temporal label; and
a zoom control configured to allow a user to modify the period of time spanned by the temporal labels;
in response to input from a user, selecting a first one or more of the plurality of identifiers and, for each of the first one or more of the plurality of identifiers, associating the respective series associated with the identifier with the left axis or the right axis;
in response to input from a user, selecting a second one or more of the plurality of identifiers;
in response to input from the user, presenting on the electronic display device a graphical display that comprises:
a first graph that comprises a first horizontal axis indicating a time interval; a first left vertical axis indicating a first range of values; a first right vertical axis indicating a second range of values; and at least one set of symbols, each set comprising at least one symbol that corresponds to at least one of the data points comprised by the stored series represented by one of the first selected one or more identifiers; wherein the horizontal placement of each symbol indicates the temporal information associated with the corresponding data point, the vertical placement of each symbol indicates the value associated with the corresponding data point, and, for each set of symbols comprised by the first graph, all symbols within the set correspond to data points comprised by the same stored series; and
a second graph that comprises a second horizontal axis indicating a time interval; a second vertical axis indicating a third range of values; and at least one set of symbols, each set comprising at least one symbol that corresponds to at least one of the data points comprised by the stored series represented by one of the second selected one or more identifiers; wherein the horizontal placement of each symbol indicates the temporal information associated with the corresponding data point, the vertical placement of each symbol indicates the value associated with the corresponding data point, and, for each set of symbols comprised by the second graph, all symbols within the set correspond to data points comprised by the same stored series;
wherein:
the first graph is presented adjacent to the second graph and either above it or below it; and
the first horizontal axis and the second horizontal axis are aligned vertically so that any position indicating a time relative to the first horizontal axis indicates the same time relative to the second horizontal axis.
US Pat. No. 9,880,180

METHODS FOR DETECTING VITAMIN D METABOLITES BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of a vitamin D metabolite in a sample by tandem mass spectrometry, comprising:
(a) generating a protonated and dehydrated precursor ion of said vitamin D metabolite;
(b) generating one or more fragment ions of said precursor ion; and
(c) detecting the amount of one or more of said ions generated in step (a) or (b) or both and relating the detected ions to
the presence or amount of said vitamin D metabolite in said sample;

wherein said vitamin D metabolite is 25-hydroxyvitamin D3 or 25-hydroxyvitamin D2 and wherein said sample or said vitamin D metabolite is not subjected to gas chromatography prior to said ionization step
(a).

US Pat. No. 10,067,133

HEMATOPOIETIC CELL PHENOTYPING USING CIRCULATING CELL-FREE MARKERS

Quest Diagnostics Investm...

1. A method for predicting survival or remission duration in a patient with myelodysplastic syndrome (MDS) or acute myeloid leukemia (AML), the method comprising:(a) assaying a blood plasma sample from a patient diagnosed as having MDS or AML to determine the level of a circulating cell-free CD34 marker in said sample, wherein assaying comprises contacting the blood plasma sample with an antibody specific for the CD34 marker,
(b) correlating the level of circulating the cell-free CD34 marker determined in step (a) with the patient's predicted survival or remission duration, and
(c) identifying the patient as
(i) having a shorter predicted survival or remission duration if the level of the cell-free CD34 marker is determined to be greater than 10,845 U/?l, whereby a level greater than 10,845 U/?l indicates that the patient will have a shorter predicted survival or remission duration than is indicated by a level less than 10,845 U/?l; or
(ii) having a longer predicted survival or remission duration if the level of the cell-free CD34 marker is determined to be less than 10,845 U/?l, whereby a level less than 10,845 U/?l indicates that the patient will have a longer survival or remission duration than is indicated by a level greater than 10,845 U/?l.
US Pat. No. 10,066,226

RNA ISOLATION FROM SOLUBLE URINE FRACTIONS

Quest Diagnostics Investm...

1. A method for processing urine for detection of RNA expression associated with a benign prostate hyperplasia or prostate cancer, comprising:a) separating urine sediment from the soluble urine fraction of a urine sample obtained from an individual suspected of having cancer, wherein RNA associated with the benign prostate hyperplasia or prostate cancer is present in the soluble urine fraction;
b) concentrating the soluble urine fraction by ultrafiltration to produce a soluble urine concentrate, wherein the volume of the soluble urine concentrate is reduced at least 50% from the original urine volume and said ultrafiltration comprises a filter having a nominal molecular weight limit of about 3,000 daltons; and
c) detecting the RNA associated with a benign prostate hyperplasia or prostate cancer in the soluble urine concentrate.

US Pat. No. 9,964,546

C PEPTIDE DETECTION BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of C peptide in a sample by tandem mass spectrometry, the method comprising:(a) subjecting a sample suspected of containing C peptide to solid phase extraction (SPE) to obtain a fraction enriched in C peptide;
(b) subjecting the enriched C peptide to an ionization source under conditions suitable to generate one or more C peptide ions detectable by mass spectrometry;
(c) determining the amount of one or more C peptide ions by tandem mass spectrometry;
wherein the amount of ions determined in step (c) is related to the amount of a C peptide in said sample.
US Pat. No. 9,957,574

BCR-ABL1 SPLICE VARIANTS AND USES THEREOF

QUEST DIAGNOSTICS INVESTM...

1. A method of detecting the presence of a nucleic acid encoding a 243-INS bcr-abl splice variant in a sample comprising bcr-abl nucleic acids, comprising contacting the sample with a labeled nucleic acid probe that hybridizes to the 5? junction site or the 3? junction site of the intron 6 insertion of the 243-INS bcr-abl splice variant and detecting the hybridized labeled nucleic acid probe to detect the bcr-abl splice variant.
US Pat. No. 9,915,663

THYROGLOBULIN QUANTITATION BY MASS SPECTROSCOPY

Quest Diagnostics Investm...

1. A method for determining the amount of thyroglobulin in a test sample, comprising:
(a) digesting thyroglobulin (Tg) and an added isotopically labeled thyroglobulin peptide standard in said test sample to form
Tg peptides and Tg peptide standard products, wherein the thyroglobulin peptide standard comprises SEQ ID NO: 2 (KVPESKVIFDANAPVAVRSKVPDS);

(b) ionizing the Tg peptide and the isotopically labeled internal standard; and
(c) quantifying the amount of the Tg peptide ion from step (b) by mass spectrometry; wherein the amount of the Tg peptide
ion detected is related to the amount of thyroglobulin in said test sample.

US Pat. No. 9,963,695

DIAGNOSIS OF PROSTATE CANCER

QUEST DIAGNOSTICS INVESTM...

1. A detection method comprising:(a) separating urine sediment from a soluble urine fraction of a urine sample obtained from an individual suspected of having prostate cancer;
(b) concentrating the soluble urine fraction by ultrafiltration to produce a soluble urine concentrate, wherein the volume of soluble urine concentrate is reduced at least 50% from the original urine volume;
(c) detecting the amount of RNA from each of heat shock 60 kDa protein 1 (HSPD1), inosine monophosphate dehydrogenase 2 (IMPDH2), PDZ and LIM domain 5 (PDLIM5), and UDP-N-acetylglucosamine pyrophosphorylase 1 (UAP1) in the concentrated soluble urine fraction.
US Pat. No. 9,957,573

EML4-ALK TRANSLOCATIONS IN LUNG CANCER

QUEST DIAGNOSTICS INVESTM...

1. A kit for detecting a EML4-ALK fusion mutation in a nucleic acid sample comprising at least one oligonucleotide comprising the sequence of SEQ ID NOs: 27 or 28, or complements thereof, wherein the at least one oligonucleotide is labeled with a reporter dye.

US Pat. No. 9,929,985

SYSTEMS AND METHODS FOR ELECTRONICALLY DISTRIBUTING INFORMATION

Quest Diagnostics Investm...

1. A computerized method for electronic distribution of information, the method being performed by a computer system that comprises one or more processors and one or more interfaces operatively connected to at least one of the processors, the method comprising:receiving through at least one of the interfaces first information from a recipient that uniquely identifies a first device associated with the recipient, the first information being received automatically as a consequence of input from the recipient and without additional human intervention;
receiving through at least one of the interfaces second information from the recipient that uniquely identifies a second device associated with the recipient, the second information being received automatically as a consequence of input from the recipient and without additional human intervention, the second device being different from the first device, and the second information being received separately from the first information;
receiving through at least one of the interfaces third information from the recipient that comprises one or more criteria according to which messages are to be sent to the first device, the second device, or both, the third information being received automatically as a consequence of input from the recipient and without additional human intervention;
receiving through at least one of the interfaces a message from a sender that comprises text, a reference to additional information, and information that identifies the recipient, the text, the reference, and the identifying information each being distinct from each other;
responsive to receipt of the message, executing instructions on at least one of the processors to identify a device or devices for receipt of the message, the identification being based on one or more of the criteria; and
transmitting through at least one of the interfaces information to cause the identified device or devices to present information comprised by the message.

US Pat. No. 9,885,724

C PEPTIDE DETECTION BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of C peptide in a sample by tandem mass spectrometry, the method comprising:
(a) subjecting a sample suspected of containing C peptide to high performance liquid chromatography (HPLC) to obtain a fraction
enriched in C peptide;

(b) subjecting the enriched C peptide to an ionization source under conditions suitable to generate one or more C peptide
ions detectable by mass spectrometry;

(c) determining the amount of one or more C peptide ions by tandem mass spectrometry, wherein said determined ions comprise
a precursor ion with a mass to charge ratio of 1007.5±0.5 and one or more fragment ions selected from the group of ions with
mass to charge ratios consisting of 927.6±0.5, 785.4±0.5, and 646.1±0.5;

wherein the amount of ions determined in step (c) is related to the amount of a C peptide in said sample.

US Pat. No. 10,309,972

C PEPTIDE DETECTION BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of C peptide in a sample by tandem mass spectrometry, the method comprising:(a) subjecting a sample suspected of containing C peptide to high performance liquid chromatography (HPLC) to obtain a fraction enriched in C peptide;
(b) subjecting the enriched C peptide to an ionization source under conditions suitable to generate one or more C peptide ions detectable by mass spectrometry;
(c) determining the amount of one or more C peptide ions by tandem mass spectrometry, wherein said determined ions comprise one or more fragment ions selected from the group of ions with mass to charge ratios consisting of 785.4±0.5 and 646.1±0.5;
wherein the amount of ions determined in step (c) is related to the amount of a C peptide in said sample.

US Pat. No. 10,209,265

CHAIN OF CUSTODY FORMS AND METHODS

Quest Diagnostics Investm...

1. A method of maintaining a chain of custody, comprising:causing a printer to print demographic information and test information upon a form that consists of a single printable sheet that is not affixed to any other sheets, the single printable sheet consisting of only a single ply that is not affixed to any other plies, the sheet comprising
at least a first portion and a second portion,
one or more seals within the second portion, each seal being removable from the sheet and capable of being affixed to a closed container so that contents of the container cannot be tampered with without either removing the seal or causing visible damage to the seal, and
an identifier that is unique to the form and is recorded at least once within the first portion and at least once upon each of the one or more seals in the second portion;
collecting from a donor at least one specimen comprising biological material, the collecting taking place under supervision of an administrator;
placing the biological material comprised by one of the specimens into a container; and
closing the container and affixing at least one of the one or more seals to the container so that contents of the container cannot be tampered with without either removing the seal or causing visible damage to the seal.
US Pat. No. 10,203,337

KISSPEPTIN-54 DETECTION BY TANDEM MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining by mass spectrometry the amount in a sample of one or more kisspeptin-54-derived peptides, said method comprising:(a) subjecting the sample to acidification;
(b) ionizing the enriched sample to produce one or more kisspeptin-54-derived peptide ions detectable by mass spectrometry, wherein said ions are selected from the group of ions with m/z of 1172.4±0.5, 977.2±0.5, 837.7±0.5, 1450.5±0.5, 1160.6±0.5, 967.3±0.5, 1143.2±0.5, 952.7±0.5, 816.9±0.5, 1112.0±0.5, 926.6±0.5, 1083.6±0.5, 903.1±0.5, 902.8±0.5, 1071.6±0.5, 1053.9±0.5, 1054.1±0.5, 878.5±0.5, 878.3±0.5, 1278.2±0.5 and 1022.9±0.5;
(c) determining the amount of one or more ions by mass spectrometry, wherein the amount of the ions is used to determine the amounts of the corresponding one or more kisspeptin-54-derived peptides in the sample.

US Pat. No. 10,194,049

PRINTING FROM A HANDHELD DEVICE VIA A REMOTE SERVER

Quest Diagnostics Investm...

1. A method of causing printing of a document in response to a request from a handheld device, the method being performed by a networked computer system that comprises one or more processors, one or more interfaces operatively coupled to at least one of the processors, and one or more databases operatively coupled to at least one of the processors, the method comprising:receiving from the handheld device, through one or more of the interfaces, a first one or more queries comprising transaction information that is related to a transaction and location information that specifies the location of the handheld device;
approving the transaction based on the transaction information;
retrieving from at least one of the databases information describing a plurality of printing systems;
based on the retrieved information and the location information, identifying a plurality of candidate printing systems;
transmitting to the handheld device, through one or more of the interfaces, information related to the candidate printing systems;
receiving from the handheld device, through one or more of the interfaces, a second one or more queries comprising information indicating selection of one of the candidate printing systems; and
transmitting to the selected printing system, through one of the interfaces, information sufficient to allow the selected printing system to render a document that is related to the transaction.
US Pat. No. 10,191,064

THYROGLOBULIN QUANTITATION BY MASS SPECTROSCOPY

Quest Diagnostics Investm...

1. A method for determining the amount of thyroglobulin in a test sample, comprising:(a) adding a thyroglobulin peptide standard in said test sample containing thyroglobulin peptides, wherein one or more valine of the thyroglobulin peptide standard is isotopically labeled with 13C, 15N;
(b) enriching said thyroglobulin peptides and thyroglobulin peptide standard from step (a);
(c) ionizing said thyroglobulin peptides and thyroglobulin peptide standard from step (b) to produce one or more thyroglobulin peptide ions and thyroglobulin peptide standard ions detectable by mass spectrometry; and
(d) detecting the amount of the ion(s) from step (c) by mass spectrometry; wherein the amount of the ion(s) detected in step (c) is related to the amount of thyroglobulin in said test sample and the amount of thyroglobulin peptide standard.

US Pat. No. 10,191,065

C PEPTIDE DETECTION BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of C peptide in a sample by high resolution/high accuracy mass spectrometry, the method comprising:(a) subjecting a sample suspected of containing C peptide to solid phase extraction (SPE) to obtain a fraction enriched in C peptide;
(b) subjecting the enriched C peptide to an ionization source under conditions suitable to generate one or more C peptide ions detectable by mass spectrometry;
(c) determining the amount of one or more C peptide ions by high resolution/high accuracy mass spectrometry;
wherein the amount of ions determined in step (c) is related to the amount of a C peptide in said sample.
US Pat. No. 10,053,738

MICRORNA PROFILING FOR DIAGNOSIS OF DYSPLASTIC NEVI AND MELANOMA

Quest Diagnostics Investm...

1. A method for distinguishing melanoma cells from normal cells in a skin lesion sample from a subject, comprising:(a) performing reverse transcriptase polymerase chain reaction (RT-PCR) to amplify nucleic acids from a skin sample isolated from a skin lesion, wherein the skin sample is suspected of containing melanoma cells;
(b) assaying the expression level of miRNAs in the skin sample by detecting the amplified nucleic acids of (a), wherein the miRNAs comprise:
i. miR-146a;
ii. hsa-let-7i;
iii hsa-let-7d; and
iv. miR-768-5p,
wherein assaying the level of expression comprises using a detectably labeled probe specific for each miRNA assayed;
(c) quantitatively comparing the levels of the miRNAs in the skin lesion sample as assayed in (b) to the levels of each corresponding miRNA in a normal skin reference sample; and
(d) identifying the skin sample as containing:
(i) melanoma cells if the levels of each of the-miRNAs in the skin sample differs from the normal skin reference sample, wherein miR-146a, hsa-let-7i, and hsa-let-7d are increased and the level of miR-768-5p is decreased relative to the normal skin reference sample; or
(ii) no melanoma cells if the levels of each of the miRNAs in the skin sample do not differ from the normal skin reference sample.
US Pat. No. 9,988,689

MICRORNA PROFILING FOR DIAGNOSIS OF DYSPLASTIC NEVI AND MELANOMA

QUEST DIAGNOSTICS INVESTM...

1. A kit for distinguishing melanoma cells from dysplastic nevi cells in a skin lesion sample from a subject comprising primer pairs that can be used for amplifying at least two miRNA selected from the group consisting of:i. miR-150;
ii. miR-149-star;
iii. miR-1308;
iv. miR-191;
v. miR-1228-star;
vi. ENSG00000199411_s;
vii. miR-1268;
viii. miR-923;
ix. miR-23a;
x. miR-132;
xi. miR-1207.5p;
xii. miR-342.3p;
xiii. U38B; and
xiv. miR-155,wherein at least one primer of each primer pair is detectably labeled.

US Pat. No. 10,325,067

STATISTICAL QUALITY CONTROL OF MEDICAL LABORATORY RESULTS

QUEST DIAGNOSTICS INVESTM...

1. A method of measuring respective values of an analyte in each of a plurality of samples of biological material, each sample being obtained at a respective time on a respective date from a respective one of a plurality of patients, the method being performed by a computer system that comprises one or more processors, one or more computer-readable storage media operatively coupled to at least one of the processors, and one or more interfaces operatively coupled to at least one of the processors, and the method comprising:receiving through at least one of the interfaces a plurality of prior values, each prior value being a measurement of a respective value of the analyte in a respective member of a population comprising a plurality of individuals;
dividing the population into a plurality of subpopulations by applying one or more criteria to the plurality of individuals, each subpopulation comprising a plurality of the plurality of individuals;
associating each of the prior values with a respective one of the subpopulations by applying the one or more criteria to the respective individual in whom the respective prior value was measured;
for each subpopulation, determining that a statistical property of the prior values associated with that subpopulation varies according to a time-periodic function, the time-periodic function being a mathematical function such that for all times t and a fixed time interval T, the value of the function at time t equals the value of the function at time t+T;
subdividing the interval T into a plurality of subintervals;
for each subpopulation and for each subinterval, based on the determined variation of the statistical property of the prior values associated with that subpopulation, calculating a reference range for the analyte that applies to the subpopulation and the subinterval;
receiving through at least one of the interfaces a plurality of values, each value being a measurement of the analyte in a respective one of the plurality of samples, the values representing respective results of an assay performed on the samples as a batch;
for each sample, determining that the sample was obtained within a particular subinterval and that the patient from whom the sample was obtained is a member of a respective subpopulation;
for each sample, identifying an applicable reference range of values based on the subinterval within which the sample was obtained and the subpopulation of which the patient from whom the sample was obtained is a member;
calculating a single quality-control grade for the batch as a whole based on, for each sample, comparing the respective measured value with the respective applicable reference range for that sample; and
based on the quality-control grade, determining to do exactly one of: 1) releasing the values for diagnostic use, and 2) repeating the assay on the samples.
US Pat. No. 10,308,680

MAGNETIC SEPARATION OF LIPOPROTEINS USING DEXTRAN SULFATE

QUEST DIAGNOSTICS INVESTM...

1. A method for purifying lipoproteins suitable for differential charged-particle mobility analysis of lipoprotein class and subclass, said method not including density gradient centrifugation; said method comprising:combining glycine with a sample comprising lipoproteins to provide a glycine mixture;
adding one or more cations and dextran sulfate to the glycine mixture to form a precipitation mixture;
contacting the precipitation mixture with an insoluble solid support, wherein one or more classes or subclasses of lipoproteins from said sample precipitate from said mixture to form precipitated lipoproteins onto said insoluble solid support;
separating said precipitated lipoproteins on said insoluble solid support from said mixture; and
removing said precipitated lipoproteins from said insoluble solid support;
wherein said precipitated lipoproteins on said insoluble solid support comprise HDL and one or more selected from the group consisting of LDL, Lp(a), IDL and VLDL;
wherein said insoluble solid support comprises magnetic beads and binds precipitated lipoproteins without an immunoaffinity agent; and
wherein said dextran sulfate is not biotinylated and said precipitated lipoproteins are not bound to said insoluble solid support by streptavidin.

US Pat. No. 10,309,971

THYROGLOBULIN QUANTITATION BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of thyroglobulin in a test sample, comprising:(a) enriching a digested thyroglobultin (Tg) peptide consisting of an amino acid sequence VIFDANAPVAVR (SEQ. ID NO: 4);
(b) ionizing the Tg peptide to generate one or more ions detectable by mass spectrometry
(c) quantifying the amount of the one or more Tg peptide ions from step (b) by mass spectrometry, wherein the amount of the one or more Tg peptide ions is related to the amount of thyroglobulin in said test sample.
US Pat. No. 10,242,852

MASS SPECTROMETRIC DETERMINATION OF FATTY ACIDS

Quest Diagnostics Investm...

1. A method for determining an amount of one or more very long chain fatty acids (VLCFA) and/or branched chain fatty acids (BCFA) in a sample by mass spectrometry, the method comprising:(a) subjecting the sample containing an amount of VLCFA and/or BCFA to an ionization source to generate one or more VLCFA and/or BCFA ions detectable by mass spectrometry, wherein said one or more ions comprise ions with a mass to charge ratio (m/z) selected from 339.3±0.5, 367.3±0.5, 395.4±0.5, 297.3±0.5, and 311.2±0.5;
(b) determining the amount of the one or more VLCFA and/or BCFA ions by mass spectrometry; and
(c) determining the amount of the VLCFA and/or BCFA in the sample from the amount of the one or more VLCFA and/or BCFA ions determined in step (b).
US Pat. No. 10,215,765

DETECTION OF VITAMINS A AND E BY TANDEM MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of vitamin A in a sample by tandem mass spectrometry, said method comprising:a. subjecting vitamin A from a sample to ionization under conditions suitable to produce one or more ions detectable by mass spectrometry; and
b. determining the amount of one or more of said ions by tandem mass spectrometry, wherein tandem mass spectrometry comprises fragmenting a vitamin A precursor ion having a mass to charge ratio of about 269.30±0.80 into one or more fragment ions comprising a fragment ion having a mass to charge ratio of about 105.00±0.80;
wherein the amount of the one or more fragment ions determined in step (b) is related to the amount of vitamin A in the sample.
US Pat. No. 10,175,208

ANALYSIS OF AMINO ACIDS IN BODY FLUID BY LIQUID CHROMOTOGRAPHY-MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of one or more amino acids in a sample by tandem mass spectrometry, the method comprising:(a) purifying the one or more amino acids by liquid chromatography (LC); and
(b) ionizing the one or more amino acids under conditions suitable to produce one or more ions detectable by mass spectrometry;
(c) detecting the amount of ions of the one or more amino acids by tandem mass spectrometry (MS/MS);
(d) determining the amount of the one or more amino acids in the sample from the amount of ions determined in step (c);
wherein the one or more amino acids comprise at least one amino acid selected from allo-isoleucine and phosphoserine.

US Pat. No. 10,164,926

SYSTEMS AND METHODS FOR ELECTRONICALLY DISTRIBUTING INFORMATION

Quest Diagnostics Investm...

1. A computer system for electronic distribution of information, the computer system comprising:one or more processors;
one or more interfaces operatively connected to at least one of the processors; and
one or more computer-readable storage media operatively connected to at least one of the processors and encoded with instructions that, when executed by at least one of the processors, cause the computer system at least to:
receive through at least one of the interfaces first information from a recipient that uniquely identifies a first device associated with the recipient, the first information being received automatically as a consequence of input from the recipient and without additional human intervention;
receive through at least one of the interfaces second information from the recipient that uniquely identifies a second device associated with the recipient, the second information being received automatically as a consequence of input from the recipient and without additional human intervention, the second device being different from the first device, and the second information being received separately from the first information;
receive through at least one of the interfaces third information from the recipient that comprises one or more criteria according to which messages are to be sent to the first device, the second device, or both, the third information being received automatically as a consequence of input from the recipient and without additional human intervention;
receive through at least one of the interfaces a message from a sender that comprises text, a reference to additional information, and information that identifies the recipient, the text, the reference, and the identifying information each being distinct from each other;
responsive to receipt of the message, identify a device or devices for receipt of the message, the identification being based on one or more of the criteria; and
transmit through at least one of the interfaces information to cause the identified device or devices to present information comprised by the message.

US Pat. No. 10,119,971

DETECTION APPARATUS FOR DIFFERENTIAL-CHARGED PARTICLE MOBILITY ANALYZER

QUEST DIAGNOSTICS INVESTM...

1. An apparatus for differential-charged particle mobility analysis and detection of fluorescence, the apparatus comprising:one or more pumps adapted to transport through a capillary;
an ionizer adapted to charge particles of a fluorophore labeled sample as the sample flows within a capillary;
a differential-charged particle mobility analyzer adapted to:
receive a sample of fluorophore labeled charged particles from an ionizer; and
perform a differential-charged particle mobility analysis on a sample of charged particles; and
a fluorescence detection system adapted to detect fluorescence of the charged particles.

US Pat. No. 10,106,857

NUCLEIC ACID DETECTION COMBINING AMPLIFICATION WITH FRAGMENTATION

Quest Diagnostics Investm...

1. A method for diagnosing an individual with cancer by determining if an individual has a mutant nucleic acid associated with the cancer, said method comprising:a) providing a sample comprising unamplified nucleic acids from the individual, wherein said unamplified nucleic acids potentially contain the mutant nucleic acid in the presence of non-mutant nucleic acid, wherein said mutant nucleic acid lacks at least one fragmentation site present in a non-mutant nucleic acid and said mutant nucleic acid and said non-mutant nucleic acid are at least 50% identical at aligned nucleotide positions;
b) fragmenting the unamplified nucleic acids under conditions such that a subsequent amplification directed to the mutant nucleic acid results in an increased detection of said mutant nucleic acid over said non-mutant nucleic acid as compared to amplification without fragmentation;
c) combining the unamplified nucleic acids of step b) with a polymerase chain reaction (PCR) mixture;
d) amplifying the mutant nucleic acid from the unamplified nucleic acids combined with the PCR mixture in step c) with a pair of primers specific for the mutant nucleic acid, wherein one primer spans or overlaps the at least one fragmentation site present in the non-mutant nucleotide sequence but absent from the mutant nucleotide sequence, and wherein fragmentation of the non-mutant nucleotide sequence in part b) prevents hybridization of said one primer to the non-mutant nucleic acid; and
e) detecting the presence or absence of an amplification product from step d) containing the mutant nucleic acid, wherein diagnosis of cancer is determined by the presence absence or amount of amplification product containing the mutant sequence;wherein the non-mutant nucleic acid comprises a wild-type nucleotide sequence and the mutant nucleic acid comprises a mutated version of the wild-type nucleic acid sequence, and wherein step b) is performed prior to step c).
US Pat. No. 10,267,810

METHODS FOR DETECTING VITAMIN D METABOLITES BY MASS SPECTROMETRY

Quest Diagnostics Investm...

1. A method for determining the amount of a vitamin D metabolite in a sample by tandem mass spectrometry, comprising:(a) generating a protonated and dehydrated precursor ion of said vitamin D metabolite by atmospheric pressure chemical ionization (APCI);
(b) generating one or more fragment ions of said precursor ion; and
(c) detecting the amount of one or more of said ions generated in step (a) or (b) or both and relating the detected ions to the presence or amount of said vitamin D metabolite in said sample;
wherein said vitamin D metabolite is 25-hydroxyvitamin D3 or 25-hydroxyvitamin D2.
US Pat. No. 10,260,113

MULTIPLEX DETECTION ASSAY FOR INFLUENZA AND RSV VIRUSES

QUEST DIAGNOSTICS INVESTM...

1. A multiplex PCR kit consisting essentially of:(a) is a primer pair comprising two of SEQ ID NO: 13, 14, or 15, or a full complement thereof and a first probe consisting of SEQ ID NO: 16 or 40, a sequence of 16-18 nucleotides and 90% identical to SEQ ID NO: 16 or 40, or a complement thereof that is 16-18 nucleotides long; and
(b) is a primer pair comprising (i) SEQ ID NO: 17 or a full complement thereof, and (ii) SEQ ID NO: 18, or 19, or a full complement thereof, and a second probe consisting of SEQ ID NO: 20, a sequence of 21-25 nucleotides and 90% identical to SEQ ID NO: 20, or a complement thereof that is 21-25 nucleotides long; and at least one of
(c) is a primer pair comprising two of SEQ ID NO: 21, 22, or 23, or a full complement thereof and a third probe consisting of SEQ ID NO: 24, a sequence of 19-23 nucleotides and 90% identical to SEQ ID NO: 24, or a complement thereof that is 19-23 nucleotides long; or
(d) is a primer pair comprising two of SEQ ID NO: 25, 26, or 27, or a full complement thereof and a fourth probe consisting of SEQ ID NO: 28, a sequence of 28-34 nucleotides and 90% identical to SEQ ID NO: 28, or a complement thereof that is 28-34 nucleotides long;
wherein the probes are attached to one or more detectable labels selected from the group consisting of fluorescent dyes, 32P, 35S, 3H, 14C, 125I, 131I, electron-dense reagents, enzymes, colorimetric labels, magnetic labels, biotin, dioxigenin, haptens, proteins for which antisera or monoclonal antibodies are available, and ligands capable of forming a complex with a corresponding receptor.

US Pat. No. 10,169,538

GRAPHICAL PRESENTATION OF MEDICAL DATA

Quest Diagnostics Investm...

1. A computer system programmed to perform a method of presenting information graphically, the method comprising:presenting on an electronic display device a flowsheet, comprising
a plurality of identifiers, each of which textually identifies exactly one of at least a plurality of stored series of one or more related data points, wherein each data point comprises a value and temporal information, the temporal information comprising a time, a date, or a time and a date, each of the identifiers being displayed in association with one or more controls, the associated controls together permitting a user to choose between associating the series with a left axis, associating the series with a right axis, and associating the series with no axis;
a plurality of temporal labels, each temporal label indicating at least a time, a date, or a time and a date, the temporal labels together spanning a period of time;
a plurality of numeric values, each numeric value representing one or more data points from a respective exactly one of the plurality of stored series, each numeric value being displayed in visual association with the identifier that identifies the respective series from which the data points were taken, each numeric value further being displayed in visual association with exactly one of the temporal labels, no more than one of the numeric values that are associated with a respective identifier being associated with any respective temporal label; and
a zoom control configured to allow a user to modify the period of time spanned by the temporal labels;
in response to input from a user, selecting a first one or more of the plurality of identifiers and, for each of the first one or more of the plurality of identifiers, associating the respective series associated with the identifier with the left axis or the right axis;
in response to input from a user, selecting a second one or more of the plurality of identifiers;
in response to input from the user, presenting on the electronic display device a graphical display that comprises:
a first graph that comprises a first horizontal axis indicating a time interval; a first left vertical axis indicating a first range of values; a first right vertical axis indicating a second range of values; and at least one set of symbols, each set comprising at least one symbol that corresponds to at least one of the data points comprised by the stored series represented by one of the first selected one or more identifiers; wherein the horizontal placement of each symbol indicates the temporal information associated with the corresponding data point, the vertical placement of each symbol indicates the value associated with the corresponding data point, and, for each set of symbols comprised by the first graph, all symbols within the set correspond to data points comprised by the same stored series; and
a second graph that comprises a second horizontal axis indicating a time interval; a second vertical axis indicating a third range of values; and at least one set of symbols, each set comprising at least one symbol that corresponds to at least one of the data points comprised by the stored series represented by one of the second selected one or more identifiers; wherein the horizontal placement of each symbol indicates the temporal information associated with the corresponding data point, the vertical placement of each symbol indicates the value associated with the corresponding data point, and, for each set of symbols comprised by the second graph, all symbols within the set correspond to data points comprised by the same stored series;
wherein:
the first graph is presented adjacent to the second graph and either above it or below it; and
the first horizontal axis and the second horizontal axis are aligned vertically so that any position indicating a time relative to the first horizontal axis indicates the same time relative to the second horizontal axis.
US Pat. No. 10,100,361

METHOD FOR DETECTING CYSTIC FIBROSIS

Quest Diagnostics Investm...

1. A method for determining the nucleotide sequence of a sample CFTR nucleic acid comprising:(a) producing an adapter-tagged amplicon library by amplifying multiple target segments of the sample CFTR nucleic acid, wherein each target segment is amplified with a pair of oligonucleotide primers, wherein at least one primer of the primer pair is selected from the group consisting of SEQ ID NOS: 3-8, 55-60, 107, 108, 141, and 142; and
(b) determining the nucleotide sequences of the target segments by sequencing the amplicons in the amplicon library using high throughput massively parallel sequencing.
US Pat. No. 10,100,365

COMPOSITIONS AND METHODS FOR DETECTING MUTATIONS IN JAK2 NUCLEIC ACID

Quest Diagnostics Investm...

1. A method for detecting a mutated JAK2 nucleic acid in a sample obtained from a human, comprising:(a) contacting the sample with a detectably labeled nucleic acid probe that specifically hybridizes to a mutant JAK2 nucleic acid but not to a wild-type JAK2 nucleic acid and comprises the mutation, wherein the mutation is a cytosine to thymine mutation at a nucleotide position 2035 of SEQ ID NO: 1; and
(b) detecting the mutant JAK2 nucleic acid when a hybrid is formed between the detectably labeled nucleic acid probe and the mutant JAK2 nucleic acid.

US Pat. No. 10,317,377

MONOLITHIC COLUMN CHROMATOGRAPHY

QUEST DIAGNOSTICS INVESTM...

1. A chromatographic method for detecting one or more analytes in a body fluid sample containing one or more other substances, said method comprising:a) applying a body fluid sample to a monolithic chromatography column comprising a monolithic sorbent having macropores and mesopores;
b) applying a first mobile phase to said monolithic chromatography column after applying said body fluid sample under conditions such that one or more analytes are retained on said monolithic chromatography column and one or more other substances present in said body fluid sample are removed, wherein said one or more retained analytes have a molecular weight of less than about 5,000 Daltons;
c) eluting said one or more retained analytes by applying a second mobile phase to said monolithic chromatography column to provide said one or more eluted analytes;
c1) directing said one or more eluted analytes to a packed capillary spray tip in-line with said monolithic chromatography column; and
d) detecting said one or more eluted analytes;
wherein the monolithic sorbent comprises a homogeneous network of nodules consisting of microparticles of polyacrylamide or functionalized polyacrylamide with an average diameter of 2 ?m.