Direct DNA amplification by PCR from oral mucosa epithelial cells

The aim of the study was to develop rapid and economical method for detection genetic diseases using PCR technology. 20 samples of buccal swabs were collected and screened for the presence of Factor V Leiden and Prothrombin 2020A mutations, which are known to be common risk alleles for venous thrombosis. We have derived conditions that enable specific and reproducible amplification of segments of Factor V and prothrombin genes directly from oral mucosa epithelial cells without previous DNA extraction as well as subsequent restriction digestion. 1 sample was found to be heterozygous for the presence of FV Leiden mutation. Our results demonstrates that use of direct PCR of oral mucosa epithelial cells offers significant advantages over the other screening methods, including both non-invasive sampling and reduction of time needed, therefore increasing the number of samples that could be handled conveniently.