Factor V Leiden Detection by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism with Mutagenic Primers in a Multiplex Reaction with Pro G20210A--A Novel Technique
- 1 January 2003
- journal article
- research article
- Published by Taylor & Francis in Hematology
- Vol. 8 (2) , 73-75
- https://doi.org/10.1080/1024533031000084222
Abstract
Factor V Leiden (FVL) R506Q and Prothrombin G20210A are clinically important genetic mutations associated with increased susceptibility to venous thrombosis. The objective of our study was to design a polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) reaction that allows simultaneous detection of these two mutations. The reaction can be used in routine diagnostic settings. We have analysed 4504 alleles for each mutation with a mutagenic primer-based PCR system with a low failure rate. The system eliminates the false positive FVL G1691A results associated with other PCR/RFLP caused by rare confounding mutations adjacent to restriction endonuclease recognition sites. This multiplex PCR/RFLP reaction is rapid, robust and dependable.Keywords
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