An automated heteroduplex assay for the Pi(A) polymorphism of glycoprotein IIb/IIIa, multiplexed with two prothrombotic genetic markers.
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.
O'Connor
F
F
Department of Clinical Pharmacology, Royal College of Surgeons in Ireland, Dublin.
Fitzgerald
D J
DJ
Murphy
R P
RP
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
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)