title: An automated heteroduplex assay for the Pi(A) polymorphism of glycoprotein IIb/IIIa, multiplexed with two prothrombotic genetic markers.
abstract: Screening for the PIA polymorphism has been made faster and simpler with the advent of heteroduplex technology. Simultaneous screening for three common prothrombotic polymorphisms pl(A), factor V Leiden, and MTHFR(C677T) has been achieved with multiplex heteroduplex analysis. We describe a quick and simple method for PlA heteroduplex probe production. The probe was multiplexed with heteroduplex probes for MTHFR(C677T) and factor V Leiden polymorphisms in a one tube assay, allowing rapid automated genotyping of all three. This automated multiplex assay was applied to a cohort of 165 patients and showed excellent correlation with gel-based assays, both PAGE and RFLP. This approach will facilitate the analysis of multiple polymorphisms in complex disease in large populations.
authors.affiliation: Department of Clinical Pharmacology, Royal College of Surgeons in Ireland, Dublin.
authors.forename: F
authors.initials: F
authors.lastname: O'Connor
authors.forename: D J
authors.initials: DJ
authors.lastname: Fitzgerald
authors.forename: R P
authors.initials: RP
authors.lastname: Murphy
mesh_terms: D015331:Cohort Studies; D001330:Electronic Data Processing; D019075:Electrophoresis, Capillary; D004591:Electrophoresis, Polyacrylamide Gel; D005165:Factor V / Q000235:genetics*; D005787:Gene Frequency; D005819:Genetic Markers; D020180:Heteroduplex Analysis / Q000379:methods*; D006801:Humans; D042965:Methylenetetrahydrofolate Reductase (NADPH2); D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000097:blood / Q000235:genetics*; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D005819:Genetic Markers; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; C095381:factor V Leiden; D005165:Factor V; D000587:Oxidoreductases Acting on CH-NH Group Donors; D042965:Methylenetetrahydrofolate Reductase (NADPH2)