An Automated Heteroduplex Assay for the Pl^A Polymorphism of Glycoprotein IIb/IIIa, Multiplexed with Two Prothrombotic Genetic Markers^[*]

Summary

Screening for the Pl^A 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 Pl^A 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.

Keywords

PlA - factor V Leiden - thermolabile MTHFR polymorphisms - automated heteroduplex assay

^* Supported by grants from the Wellcome Trust, the Higher Education Authority (HEA) and the Health Research Board (HRB) of Ireland. Dr. Murphy was a HRB Postdoctoral Fellow during the performance of this work.

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