A bovine protocol for training professionals in preimplantation genetic diagnosis using polymerase chain reaction.
OBJECTIVE
To develop a bovine protocol for training in preimplantation genetic diagnosis (PGD) using PCR.


METHODS
Randomized study.


METHODS
Human reproduction PCR laboratory.


METHODS
Cow ovaries obtained from slaughterhouses.


METHODS
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.


METHODS
Establishment of a possible training protocol.


RESULTS
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 successful in both embryos and blastomeres. Sex was identical in all these cases.


CONCLUSIONS
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.
Almodin
Carlos Gilberto
CG
Materbaby--Reprodução Humana e Genética, Maringá, Paraná, Brazil. almodin@materbaby.com.br
Moron
Antonio Fernandes
AF
Kulay
Luiz
L
Minguetti-Câmara
Vânia Cibele
VC
Moraes
Andréa Cristina
AC
Torloni
Maria Regina
MR
D000818:Animals; D002417:Cattle; D004517:Education, Professional / Q000379:methods; D004622:Embryo, Mammalian / Q000502:physiology; D005260:Female; D006801:Humans; D017056:Medical Laboratory Personnel / Q000193:education*; D016133:Polymerase Chain Reaction / Q000379:methods*; D019836:Preimplantation Diagnosis / Q000379:methods*; D019849:Sex Determination Processes; D013663:Teaching / Q000379:methods