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[Preclinical study of preimplantation genetic diagnosis in animal]
1Center of Reproductive Medicine, Changhai Hospital, Second Military Medical University, Shanghai 200433, China.
Objective:
To perform preimplantation genetic diagnosis (PGD) in transgenic mice.
Methods:
Single blastomere from murine heterozygous pre-embryo at 4 or 8-cell stage was sampled by microbiopsy for detection of the human gene delta F508 mutation of cystic fibrosis and biopsied pre-embryo was transferred to Swiss mice foster mothers. delta F508 mutation detection on the single blastomere was performed with nested polymerase chain reaction(PCR).
Results:
(1) Significant difference was not observed statistically for the in vitro hatching rate (n = 102), the birth rate (n = 79), birth weight and organ weight (n = 133) at 3 weeks after births between mice born from biopsied embryos and controls. (2) Thirty-two nested PCR were performed for the diagnosis of the cystic fibrosis delta F508 mutation, with 100.0% specificity and 100.0% sensitivity.
Conclusions:
Microbiopsy of pre-embryos with 4 or 8-cells did not alter their viability and further development. The technique of nested PCR on a single blastomere for the detection of cystic fibrosis delta F508 mutation is reliable.