Single cell detection of inherited retinoblastoma predisposition

Retinoblastoma susceptibility is an autosomal dominantly inherited cancer p redisposition which also confers a life-long increased risk for various non -ocular malignancies. We developed a protocol for single cell detection of this disorder which enables its preimplantation genetic diagnosis as an alt ernative to prenatal diagnosis with attendant pregnancy termination. The pr esented method detects the underlying mutation of the disease, a linked int ragenic polymorphism (p88PR0.6) and an independent marker (D21S1411) for ge netic fingerprinting allowing detection of contamination. The strategy is b ased on the combination of nested tripler polymerase chain reaction, single strand conformation polymorphism analysis by conventional polyacrylamide g el electrophoresis and fragment size determination with automated laser flu orescence. Copyright (C) 1999 John Wiley & Sons, Ltd.