KASP-PCR method to screen thrombophilia genetic risk factors

Thrombophilia is defined as the willingness of blood to clot easily in a situation of imbalances between fibrinolysis and coagulation. It is classified as inherited and acquired thrombophilia. Several studies revealed that the inherited thrombophilia is strongly associated with single nucleotide polymorphisms (SNP) or deletions on certain genes, such as FV Leiden (rs6025), MTHFR1 (rs1801133), MTHFR2 (rs1801131), Serpine-1 (rs1799768), and Factor II (rs1799963). This study aims to develop an SNP detection panel based on Kompetitive Allele Specific Primer-polymerase chain reaction (KASP-PCR) technique. Results revealed that 86.5% of susceptible patients (n = 111) contain at least one mutation; while seven of them harbor three thrombophilia-associated mutations simultaneously. A clear allelic discrimination was observed for all tested samples. The prevalence of each mutation among different countries and ethnic groups are in line with the findings of this study. Rather than expensive and time-consuming approaches, the current assay enables the cost-effective advantage of the KASP-PCR, which reduces the diagnostic cost with a fast and convenient way. After clinical validation and approval, it can be used in hospitals, research centers, and diagnostic laboratories to determine the genetic susceptibility of individuals to thrombosis and for research purposes.