A multiplex assay for detecting genetic variations in CYP2C9, VKORC1, and GGCX involved in warfarin metabolism.

Abstract:

BACKGROUND:Patients differ in responses to warfarin, which is commonly prescribed to treat thromboembolic events. Genetic variations in the cytochrome P450, family 2, subfamily C, polypeptide 9 (CYP2C9), vitamin K epoxide reductase complex, subunit 1 (VKORC1), and gamma-glutamyl carboxylase (GGCX) genes have been shown to contribute to impaired metabolism of warfarin. METHODS:We designed a custom multiplex single-nucleotide polymorphism (SNP) panel to interrogate the CYP2C9 *2, *3, VKORC1 (-1639G-->A), and GGCX (1181T-->G) alleles simultaneously in a single sample by use of single-base extension and capillary electrophoresis after genomic DNA extraction and PCR amplification. RESULTS:Our assay successfully detected various genotypes from known controls and 24 unknown samples. It was found to be 100% concordant with sequencing results. CONCLUSIONS:Our multiplexed SNP panel can be successfully used in genotyping of patient blood samples. Results can be combined with other clinical parameters in an algorithm for warfarin dosing. These data provide a proof-in-principle of multiplexed SNP analysis using rapid single-base extension and capillary electrophoresis, and warrant additional validation using a larger cohort of patient samples.

journal_name

Clin Chem

journal_title

Clinical chemistry

authors

Rai AJ,Udar N,Saad R,Fleisher M

doi

10.1373/clinchem.2008.118497

subject

Has Abstract

pub_date

2009-04-01 00:00:00

pages

823-6

issue

4

eissn

0009-9147

issn

1530-8561

pii

clinchem.2008.118497

journal_volume

55

pub_type

杂志文章
  • Low apparent creatine kinase activity and prolonged lag phases in serum of patients with metastatic disease: elimination by treatment of sera with sulfhydryl agents.

    abstract::We find lag phases exceeding 20 min in measuring creatine kinase activity, by using the kinetic creatin phosphate leads to creatine assay, in sera of some patients with carcinoma metastatic to the liver. Such long lag phases are accompanied by a decreased apparent enzyme activity. These problems are eliminated by addi...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Bruns DE,Morgan WS,Davis JE,Ladenson JH

    更新日期:1976-11-01 00:00:00

  • "Ultrasensitive" silver stains: their use exemplified in the study of normal human cerebrospinal fluid proteins separated by two-dimensional electrophoresis.

    abstract::We have developed a series of photochemical silver stains that are more than 100 times as sensitive as the Coomassie Blue stains. These stains facilitate the quantitative estimation of proteins in two-dimensional gel patterns. Recently, we discovered that by using light (photodevelopment) to catalyze the reduction of ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Merril CR,Harrington MG

    更新日期:1984-12-01 00:00:00

  • Induced gene transcription: implications for biomarkers.

    abstract::Numerous xenobiotics regulate cellular functions by altering transcription of target genes. Use of sensitive and specific biomarkers based on gene transcript levels may help clarify the shape of the dose-response curve in the low-dose region associated with human exposures to environmental concentrations of chemicals....

    journal_title:Clinical chemistry

    pub_type: 杂志文章,评审

    doi:

    authors: Sewall CH,Bell DA,Clark GC,Tritscher AM,Tully DB,Vanden Heuvel J,Lucier GW

    更新日期:1995-12-01 00:00:00

  • Improved biuret procedure for routine determination of urinary total proteins in clinical proteinuria.

    abstract::This communication describes and evaluates an improved routine methodology for quantitating clinical proteinuria. Based on investigations of Piscator and of Savory et al., a modified Tsuchiya's reagent (ethanolic HCI-phosphotungstic acid) is used to precipitate proteins at 56 degrees C, followed by biuret spectrophoto...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Rice EW

    更新日期:1975-03-01 00:00:00

  • Analytical performance evaluation of EMIT II monoclonal amphetamine/methamphetamine assay: more specificity than EMIT d.a.u. monoclonal amphetamine/methamphetamine assay.

    abstract::We evaluated a new EMIT II monoclonal amphetamine/methamphetamine assay for screening human urine by comparing it with the EMIT d.a.u. monoclonal amphetamine/methamphetamine assay and a fluorescence polarization assay. The EMIT II assay has a cutoff of 1 mg/L d-methamphetamine. The EMIT II and EMIT d.a.u. assays were ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Dasgupta A,Saldana S,Kinnaman G,Smith M,Johansen K

    更新日期:1993-01-01 00:00:00

  • The scope of mass spectrometry in clinical chemistry.

    abstract::Several aspects of mass spectrometry and gas chromatography-mass spectrometry are reviewed, including ionization methods, repetitive scanning, selected ion monitoring, and stable isotope measurement. I discuss the application of these and other mass spectrometry methods to the investigation of compound classes relevan...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Lawson AM

    更新日期:1975-06-01 00:00:00

  • Validated gas chromatographic-negative ion chemical ionization mass spectrometric method for delta(9)-tetrahydrocannabinol in sweat patches.

    abstract:BACKGROUND:A sensitive gas chromatography-negative ion chemical ionization mass spectrometry (GC/MS-NICI) method was developed and validated for the measurement of Delta(9)-tetrahydrocannabinol (THC) in human sweat patches. METHODS:THC-d(0) and THC-d(3) were added to worn blank sweat patches (PharmChek; PharmChem Inco...

    journal_title:Clinical chemistry

    pub_type: 临床试验,杂志文章

    doi:10.1373/clinchem.2004.034868

    authors: Saito T,Wtsadik A,Scheidweiler KB,Fortner N,Takeichi S,Huestis MA

    更新日期:2004-11-01 00:00:00

  • Advanced glycosylation end products in the mesenteric artery.

    abstract::We measured advanced glycosylation end products in the mesenteric artery of 37 patients (ages 29-82 years), 34 of whom were nondiabetic. Samples of arterial tissue were obtained during bowel resectioning. Advanced glycosylation end products were measured as collagen-linked fluorescence (excitation wavelength 370 nm, e...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: MacDonald E,Lee WK,Hepburn S,Bell J,Scott PJ,Dominiczak MH

    更新日期:1992-04-01 00:00:00

  • Clinical utility and validation of emerging biochemical markers for mammary adenocarcinoma.

    abstract::The clinical utilities of the emerging biochemical markers for mammary adenocarcinoma CA 15-3, CA 549, CA M26, CA M29, and MCA (mucin-like carcinoma-associated antigen) were assessed by a formal rating according to six desirable marker characteristics. All five indicators similarly have good specificities but limited ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章,评审

    doi:

    authors: Werner M,Faser C,Silverberg M

    更新日期:1993-11-01 00:00:00

  • Toward absolute methods in clinical chemistry: application of mass fragmentography to high-accuracy analyses.

    abstract::We review recently developed reference methods based on mass fragmentography (specific ion monitoring) with use of isotope-labeled internal standard, for determination of cholesterol, triglycerides, urea, glucose, cortisol, progesterone, and testosterone. With an optimal ratio between standard and material to be deter...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Björkhem I,Blomstrand R,Lantto O,Svensson L,Ohman G

    更新日期:1976-11-01 00:00:00

  • Composition of serum very-low-density and high-density lipoproteins in diabetes.

    abstract::We examined the cholesterol/protein ratio and the apoprotein composition of serum lipoproteins in a randomly selected population of maturity-onset diabetics and in a group of nondiabetics of similar age. We found no differences in cholesterol distribution between the groups as a whole, but diabetics with above-normal ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Gabor J,Spain M,Kalant N

    更新日期:1980-08-01 00:00:00

  • Stable-isotope dilution liquid chromatography-electrospray injection tandem mass spectrometry method for fast, selective measurement of S-adenosylmethionine and S-adenosylhomocysteine in plasma.

    abstract:BACKGROUND:It has been postulated that changes in S-adenosylhomocysteine (AdoHcy), a potent inhibitor of transmethylation, provide a mechanism by which increased homocysteine causes its detrimental effects. We aimed to develop a rapid and sensitive method to measure AdoHcy and its precursor S-adenosylmethionine (AdoMet...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2004.046995

    authors: Gellekink H,van Oppenraaij-Emmerzaal D,van Rooij A,Struys EA,den Heijer M,Blom HJ

    更新日期:2005-08-01 00:00:00

  • A more sensitive automated method for determination of ornithine carbamoyltransferase activity in human serum.

    abstract::Ornithine carbamoyltransferase (EC 2.1.3.3) activity is a sensitive, specific indicator of hepatocellular injury. This paper describes development of an improved automated procedure for measurement of this activity. Triethanolamine-ethylenediaminetetraacetate is used as a buffer, and activity is determined by measurin...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Clayson KJ,Fine JS,Strandjord PE

    更新日期:1975-05-01 00:00:00

  • Detection of mucopolysaccharidosis type II by measurement of iduronate-2-sulfatase in dried blood spots and plasma samples.

    abstract:BACKGROUND:Mucopolysaccharidosis type II (MPS II) is a lysosomal storage disorder related to a deficiency in the enzyme iduronate-2-sulfatase (IDS). Clinical trials of enzyme replacement therapy are in progress, but effective treatment will require screening assays to enable early detection and diagnosis of MPS II. Our...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2005.061838

    authors: Dean CJ,Bockmann MR,Hopwood JJ,Brooks DA,Meikle PJ

    更新日期:2006-04-01 00:00:00

  • Alanine aminotransferase and aspartate aminotransferase measurements with two automated analyzers, SMAC and the ABA-100, compared.

    abstract::Measurements of alanine aminotransferase and aspartate aminotransferase with the SMAC were evaluated for correlation with the ABA-100, precision, linearity, and carryover. We assayed 200 specimens with normal and abnormal aminotransferase activities with both the SMAC (y) and the ABA-100 (X). Linear regression analysi...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Purcell GV,Behenna DB,Walsh PR

    更新日期:1979-05-01 00:00:00

  • Newborn screening for cystic fibrosis by use of a multiplex immunoassay.

    abstract:BACKGROUND:Since its beginnings, newborn screening for cystic fibrosis (CF) using an assay for immunoreactive trypsinogen (IRT) has been plagued by a high rate of false-positive results (screen positive, diagnosis negative), despite attempts to reduce this rate by use of altered cutoffs and second-tier DNA testing. IRT...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2009.132480

    authors: Lindau-Shepard BA,Pass KA

    更新日期:2010-03-01 00:00:00

  • Clinical and genetic correlates of growth differentiation factor 15 in the community.

    abstract:BACKGROUND:Growth differentiation factor 15 (GDF15), a stress-responsive cytokine produced in cardiovascular cells under conditions of inflammation and oxidative stress, is emerging as an important prognostic marker in individuals with and without existing cardiovascular disease (CVD). We therefore examined the clinica...

    journal_title:Clinical chemistry

    pub_type: 杂志文章,meta分析

    doi:10.1373/clinchem.2012.190322

    authors: Ho JE,Mahajan A,Chen MH,Larson MG,McCabe EL,Ghorbani A,Cheng S,Johnson AD,Lindgren CM,Kempf T,Lind L,Ingelsson E,Vasan RS,Januzzi J,Wollert KC,Morris AP,Wang TJ

    更新日期:2012-11-01 00:00:00

  • Outliers as a cause of false cardiac troponin results: investigating the robustness of 4 contemporary assays.

    abstract:BACKGROUND:It is important that cardiac troponin be measured accurately with a robust method to limit false results with potentially adverse clinical outcomes. In this study, we characterized the robustness of 4 analytical platforms by measuring the outlier rate between duplicate results. METHODS:We measured cardiac t...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2010.159830

    authors: Pretorius CJ,Dimeski G,O'Rourke PK,Marquart L,Tyack SA,Wilgen U,Ungerer JP

    更新日期:2011-05-01 00:00:00

  • RNA chip: quality assessment of RNA by microchannel linear gel electrophoresis in injection-molded plastic chips.

    abstract::Two major components of rRNA (18S and 28S rRNA) were separated by electrophoresis in injection-molded acrylic chips with a microchannel 100 microm in width, 40 microm in depth, and with 1 cm of separation distance. Microchannels were filled with 4 g/L hydroxypropylmethylcellulose as sieving polymer and 5 mg/L ethidium...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Ogura M,Agata Y,Watanabe K,McCormick RM,Hamaguchi Y,Aso Y,Mitsuhashi M

    更新日期:1998-11-01 00:00:00

  • Ten commercial kits compared for assay of thyrotropin in the normal and thyrotoxic range.

    abstract::I developed a standard procedure for assessing sensitivity, intra-assay precision, parallelism of sample dilutions, and assay drift, using a single trial assay kit. I used this to compare the performance of 10 commercial kits with an in-house radioimmunoassay for human thyrotropin. No one kit stood out as clearly supe...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Evans MC

    更新日期:1988-01-01 00:00:00

  • Diagnostic accuracy of pancreatic enzymes evaluated by use of multivariate data analysis.

    abstract::We analyzed pancreatic enzyme data from 508 patients with suspected pancreatitis by neural network analysis, by an Expert multirule generation protocol, and by receiver-operator characteristic (ROC) curve analysis of a single test result. Neural network analysis showed that use of lipase provided the best means for di...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Kazmierczak SC,Catrou PG,Van Lente F

    更新日期:1993-09-01 00:00:00

  • Human kallikrein gene 5 (KLK5) expression by quantitative PCR: an independent indicator of poor prognosis in breast cancer.

    abstract:BACKGROUND:KLK5 is a newly discovered human kallikrein gene. Many kallikrein genes have been found to be differentially expressed in various malignancies, and prostate-specific antigen (PSA; encoded by the KLK3 gene) is the best tumor marker for prostate cancer. Like the genes that encode PSA and other kallikreins, the...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Yousef GM,Scorilas A,Kyriakopoulou LG,Rendl L,Diamandis M,Ponzone R,Biglia N,Giai M,Roagna R,Sismondi P,Diamandis EP

    更新日期:2002-08-01 00:00:00

  • Applications of recombinant DNA to pathologic diagnosis.

    abstract::Recombinant DNA techniques are contributing to the understanding of the pathogeneses of genetic, neoplastic, and viral diseases, and are used in the diagnosis of certain genetic and viral diseases. Such techniques will have wider application in the future and will play an increasing role in the clinical laboratory. Th...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Sauls CD,Caskey CT

    更新日期:1985-06-01 00:00:00

  • Meconium nicotine and metabolites by liquid chromatography-tandem mass spectrometry: differentiation of passive and nonexposure and correlation with neonatal outcome measures.

    abstract:BACKGROUND:Meconium analysis is a diagnostically sensitive and objective alternative to maternal self-report for detecting prenatal tobacco exposure. Nicotine and metabolite disposition in meconium is poorly characterized, and correlation of analytes' concentrations with neonatal outcomes is unexplored. Our objectives ...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:10.1373/clinchem.2008.109173

    authors: Gray TR,Magri R,Shakleya DM,Huestis MA

    更新日期:2008-12-01 00:00:00

  • Five creatine kinase isoenzymes in serum of a patient with severe heart disease.

    abstract::Abnormal creatine kinase (CK) isoenzyme patterns were observed in the serum of a 64-year-old woman with severe heart disease. Agarose electrophoresis revealed the presence of all the usual CK isoenzymes (MM, MB, and BB) plus an extra band between MM and MB. Total serum CK activity was within the normal range. Within 2...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Csako G,Papadopoulos NM,Jett GK,McIntosh CL

    更新日期:1982-10-01 00:00:00

  • Analytical performance of EMIT cyclosporine assay evaluated.

    abstract::We evaluated the EMIT Cyclosporine Assay (Syva Co., Palo Alto, CA), using the Cobas-Mira analyzer to assess the precision, accuracy, and analytical recovery from whole-blood samples supplemented with cyclosporine. We also performed comparative analysis of whole-blood samples containing cyclosporine from liver and kidn...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Dasgupta A,Saldana S,Desai M

    更新日期:1991-12-01 00:00:00

  • Critical assessment of phospholipid measurement in amniotic fluid.

    abstract::We assessed several methods of inorganic phosphate assay for their suitability in estimating phospholipids in digested extracts of amniotic fluids. The extraction and digestion procedures used for phospholipids from amniotic fluid were also examined critically. The effect of contamination by blood or obstetric cream h...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Badham LP,Worth HG

    更新日期:1975-09-01 00:00:00

  • Meaning of serum free-thyroxin values in nonthyroidal illnesses: seven methods compared.

    abstract::Serum free thyroxin (FT4) was determined in 40 patients with various nonthyroidal illnesses. We studied seven methods: (1) a free thyroxin index calculated from total T4 and triiodothyronine resin uptake; (2) a free T4 index determined by enzyme inhibitor assays (Abbott's "Tetrazyme" and "Thyrozyme"); (3) a free T4 in...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Chan DW,Waud JM,Taylor E,Stem J,Drew H,Oropeza M,Sucupira MS

    更新日期:1983-12-01 00:00:00

  • Measurement of elastase and cysteine proteinases in synovial fluid of patients with rheumatoid arthritis, sero-negative spondylarthropathies, and osteoarthritis.

    abstract::Synovial fluid samples were collected from 45 patients with rheumatoid arthritis, spondylarthropathy, or osteoarthritis, to study their content of elastase (EC 3.4.21.37) and of cysteine proteinases (EC 3.4.22.1, 3.4.22.15). We measured both elastase complexed with alpha 1-proteinase inhibitor and elastase activity to...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Huet G,Flipo RM,Richet C,Thiebaut C,Demeyer D,Balduyck M,Duquesnoy B,Degand P

    更新日期:1992-09-01 00:00:00

  • Rapid liquid chromatography-tandem mass spectrometry method for routine analysis of cyclosporin A over an extended concentration range.

    abstract:BACKGROUND:Cyclosporin A (CsA) is commonly measured by immunoassay techniques that have a limited analytical range. The consequence of this is that low CsA concentrations that may be clinically significant are difficult to measure and that high concentrations require sample dilution, which introduces error and increase...

    journal_title:Clinical chemistry

    pub_type: 杂志文章

    doi:

    authors: Keevil BG,Tierney DP,Cooper DP,Morris MR

    更新日期:2002-01-01 00:00:00