Mutation analysis in fatal pulmonary thromboembolism—postmortem validation study and beyond

Abstract Sudden fatal pulmonary thromboembolism (PE) is very common in Caucasians and results in over ∼ 120,000 deaths per year. There are both acquired and inherited risk factors for PE. The three most common mutations are Factor V Leiden G1691A, Prothrombin G20210A, and MTHFR C677T. We have developed an in-house molecular testing methodology using polymerase chain reaction (PCR) and automated DNA sequencing technologies. The method was validated on postmortem tissue samples, such as heart, spleen, and liver. Tissues were stored in RNAlater solution for up to 2 years. The method has also been validated on blood specimens, which were dried on staincards and stored at room temperature for up to 2 years. We obtained results for all tested specimens, including those displaying varying degrees of decomposition. The analytic sensitivity, specificity, and reproducibility show that the method is highly sensitive, and very specific for all three mutations.