Method Of Sex Identification In Preimplantation Genetic Diagnosis Of Single Gene Disorders In The Study

At present, more than 200 diseases which have been identified are X-linked,these diseases only affect male fetuses. Therefore,we developed two methods(nested PCR and primer extension pre-amplification) to diagnosis embryo's sex in preimplantation genetic diagnosis to discard male embryos and implant female embryos in uteruses. to investigate sensitivity, reliability, accuracy of nested PCR and primer extension pre-amplification in single cells sex determination, we amplified 31 single male Lymphocytes and 31 single female Lymphocytes by nested PCR and primer extension pre-amplification respectively, further,we amplified 31 single blastomeres by primer extension pre-amplification. The results: by nested PCR,96.8％(30/31) single male Lymphocytes were amplified successfully, single female Lymphocytes were all negative(0/31). By extension pre-amplification, all single Lymphocytes were amplified successfully. 96.8％(30/31) single male Lymphocytes were amplified successfully,6.5％(2/31) single female Lymphocytes were false positive; 93.5％(29/31) embryo cells were amplified successfully, 77.4％(24/31) embryo cells were diagnosised male and 16.1％(5/31) embryo cells were diagnosised female. we get the conclusions: nested PCR and primer extension pre-amplification are reliable in sexing single cells, and can be applied in PGD, moreover,extension pre-amplification can have better perspective than nested PCR.