COMPARISON OF FISH, PRINS, AND CONVENTIONAL AND FLUORESCENT PCR FOR SINGLE-CELL SEXING, SUITABILITY FOR PREIMPLANTATION GENETIC DIAGNOSIS

Purpose: Although conventional polymerase chain reaction (PCR) was the first method used for sexing in preimplantation genetic diagnosis, fl uorescent in situ hybridization (FISH) has become the method of choice . Recently two new techniques, primed in situ synthesis (PRINS) and fl uorescent PCR, have been developed. This study compares the reliabilit y and accuracy of these four techniques in single cells. Results: In b uccal cells, fluorescent PCR and FISH had similar reliability (94 and 93%) and accuracy (97 and 96%) rates. The reliability and accuracy of PRINS (91 and 25%) and conventional PCR (79 and 89%) were lower. In hu man blastomeres, FISH and flourescent PCR had similar reliability (100 %, 7/7; 95%, 190/201) rates. Accuracy rates,cere 71 % (5/7) and 99% (1 88/190)for FISH and fluorescent PCR, respecticely, however; too few bl astomeres were analyzed by FISH for meaningful comparison. However; wh en these data are compared,with published data, the method of choice f or blastomere sexing appears to be fluorescent PCR.Conclusions: Flouro scent PCR has major implications for PGD.