A bovine protocol for training professionals in preimplantation genetics diagnosis using polymerase chain reactions

Abstract

Objective: To develop a bovine protocol for training in preimplantation genetic diagnosis (PGD) using PCR.Design: Randomized study.Setting: Human reproduction PCR laboratory.Patient(s): Cow ovaries obtained from slaughterhouses.Intervention(s): the ovaries were punctured and the oocytes were matured and submitted to in vitro fertilization. On the third day after fertilization, the embryos were biopsied and 1-2 blastomeres removed. A blastomere and the rest of the embryo were submitted to PCR for sex determination.Main Outcome Measure(S): Establishment of a possible training protocol.Result(s): A total of 50 embryos and 50 biopsied blastomeres were submitted to DNA amplification for sexing. of the 50 embryos, 41 (82%) achieved successful DNA amplification and 9 (18%) did not. of the 50 biopsies, 31 (62%) amplified and 19 (38%) did not. in 27 (65.9%) of the 41 embryos with DNA amplification, sex was identified as female and in 14 (34.1%) as male. in 40 cases (80%) amplification and sex determination were in both embryos and blastomeres. Sex was identical in all these cases.Conclusion(s): This training model seems to be useful in identifying mistakes and difficulties and improving the professional's performance in the various stages of preimplantation genetic diagnosis.