First preimplantation genetic diagnosis of hereditary retinoblastoma using informative microsatellite markers. Girardet A, Hamamah S, Anahory T, Déchaud H, Sarda P, Hédon B, Demaille J, Claustres M. Mol Hum Rep 2003;9:111–116

Abstract

Retinoblastoma occurs when both alleles of the retinoblastoma gene (RB1) are inactivated. In this study two highly polymorphic microsatellite markers, RB1.20 and D13S284, located within and close to the RB1 gene, were used to clinically obtain a preimplantation genetic diagnosis for hereditary retinoblastoma. Duplex polymerase chain reactions (PCRs) were tested on more than 300 single lymphocytes from heterozygous individuals at both loci to test the accuracy and reliability of the single-cell protocol. This procedure requires a nested PCR and the analysis of fluorescently labeled PCR products on an automatic DNA sequencer. Amplification efficiency and allele dropout rates ranged from 96.17 to 98.4% and 3.7 to 5.4%, respectively. This test was found to be accurate and reliable enough to be applied to the study of human blastomeres. Subsequently, this approach was used in a preimplantation genetic diagnosis treatment cycle for a couple who already had a child affected with hereditary retinoblastoma and was found to be informative for both microsatellite markers.—Hans E. Grossniklaus