Development of a simple multiplex polymerase chain reaction for the simultaneous detection of factor V Leiden and prothrombin 20210A mutations.

BACKGROUND: The demand for thrombophilia testing at the molecular level is increasing, and consequently, the work load of the routine molecular laboratory is also increasing. Efforts to lighten the work load, economize on time, and strive for reduced costs while still maintaining quality assurance are thus necessary. METHODS AND RESULTS: A multiplex polymerase chain reaction (PCR) for the detection of factor V Leiden and prothrombin 20210A mutations was designed that enables the use of the same inexpensive restriction enzyme, controls for the digestion, and produces easily interpretable results. CONCLUSION: The use of this new multiplex PCR and digestion analysis enabled us to simultaneously perform a routine screen for factor V Leiden and prothrombin 20210A mutations.