Applicability of hot start PCR in the PCR-based preimplantation genetic diagnosis: improvement of sensitivity, specificity, and yield in multiplex nested PCR

Abstract

OBJECTIVE: Pre-implantation genetic diagnosis (PGD) should achieve a higher level of accuracy and reliability than any other method. Hereupon, hot start PCR reduces non-specific amplification and increases PCR product target yield to inhibit DNA polymerase activity during PCR reaction cycling. We have developed a test which can directly be used for preimplantation genetic diagnosis of the dystrophin gene deletion. It consists of a multiplex nested PCR using hot start PCR, allowing the amplification of four DMD exons as well as the ZFX/ZFY gene.