DNA-BASED ASSAY FORMATS FOR DETECTING THROMBOSIS FACTOR-V GENOTYPES

Substitution at factor V nucleotide 1691, the Leiden mutation, is the most common known genetic cause of venous thrombosis, DNA-based assays for the detection of factor V Leiden alleles follow common strategies , The Amplify, Interrogate, and Detect strategy includes polymerase ch ain reaction (PCR) amplification of a factor V gene fragment, interrog ation with a sequence-specific reagent, such as restriction endonuclea se or oligonucleotide probe, and detection of the results of interroga tion, The Differentially Amplify and Detect strategy uses sequence-spe cific PCR primers to selectively amplify factor V Leiden or normal all eles, followed by detection of the amplified fragments, Several assays have been validated in comparison studies and are reliable if perform ed properly. The choice of assay format depends upon laboratory experi ence and sample volume.