title: Detection of genomic polymorphisms associated with venous thrombosis using the invader biplex assay.
abstract: A multi-site study to assess the accuracy and performance of the biplex Invader assay for genotyping five polymorphisms implicated in venous thrombosis was carried out in seven laboratories. Genotyping results obtained using the Invader biplex assay were compared to those obtained from a reference method, either allele-specific polymerase chain reaction (AS-PCR), restriction fragment length polymorphism (PCR-RFLP) or PCR-mass spectrometry. Results were compared for five loci associated with venous thrombosis: Factor V Leiden, Factor II (prothrombin) G20210A, methylenetetrahydrofolate reductase (MTHFR) C677T and A1298C, and plasminogen activator inhibitor (PAI-1) 4G/5G. Of a total of 1448 genotypes tested in this study, there were 22 samples that gave different results between the Invader biplex assay and the PCR-based methods. On further testing, 21 were determined to be correctly genotyped by the Invader Assay and only a single discrepancy was resolved in favor of the PCR-based assays. The compiled results demonstrate that the Invader biplex assay provides results more than 99.9% concordant with standard PCR-based techniques and is a rapid and highly accurate alternative to target amplification-based methods.
authors.affiliation: Specialty Laboratories Inc., Santa Monica, California, USA. meeta@Pathwaydx.com
authors.forename: Madhumita
authors.initials: M
authors.lastname: Patnaik
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Dlott
authors.forename: Robert N
authors.initials: RN
authors.lastname: Fontaine
authors.forename: M T
authors.initials: MT
authors.lastname: Subbiah
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Hessner
authors.forename: Kelly A
authors.initials: KA
authors.lastname: Joyner
authors.forename: Marlies R
authors.initials: MR
authors.lastname: Ledford
authors.forename: Eduardo C
authors.initials: EC
authors.lastname: Lau
authors.forename: Cynthia
authors.initials: C
authors.lastname: Moehlenkamp
authors.forename: Jean
authors.initials: J
authors.lastname: Amos
authors.forename: Bailing
authors.initials: B
authors.lastname: Zhang
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Williams
mesh_terms: D004247:DNA / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D005165:Factor V / Q000235:genetics; D005456:Fluorescent Dyes; D005838:Genotype; D006801:Humans; D013058:Mass Spectrometry; D042965:Methylenetetrahydrofolate Reductase (NADPH2) / Q000235:genetics; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D011516:Prothrombin / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D020246:Venous Thrombosis / Q000175:diagnosis* / Q000235:genetics*
chemical_list: D005456:Fluorescent Dyes; D017395:Plasminogen Activator Inhibitor 1; C095381:factor V Leiden; D005165:Factor V; D011516:Prothrombin; D004247:DNA; D042965:Methylenetetrahydrofolate Reductase (NADPH2)