运用单细胞聚合酶链式反应进行植入前诊断的性别鉴定

目的 :建立一种高效、快速、可靠的植入前诊断的性别鉴定方法。方法 :取成人外周血单个淋巴细胞和人胚胎单卵裂球 ,采用多重荧光PCR和多重巢式PCR同时扩增性别识别位点Amelogenin和 5号染色体上的基因IL 3。结果 :男性单个淋巴细胞荧光PCR扩增效率为 72 % ,巢式PCR扩增效率为 6 4 .7% ;女性单个淋巴细胞荧光PCR扩增效率为 76 % ,巢式PCR扩增效率为 6 5 % ,两者与供体外周血gDNA扩增结果的一致性均达 10 0 %。单卵裂球荧光PCR扩增效率为 86 .9% ,巢式PCR扩增效率为 85 .7% ,来源于同一胚胎的不同单卵裂球扩增结果的一致性达 10 0 %。巢式PCR与荧光PCR的扩增效率比较无显著性差异 (P >0 .0 5 )。结论 :巢式PCR和荧光PCR各有优劣 ,但荧光PCR操作简便 ,耗时短、灵敏度高 ,为今后的发展方向

Single cell PCR for sex determination in preimplantation genetic diagnosis

Objective To develop an efficient, rapid and reliable method for sex determination in preimplantation genetic diagnosis. Methods Nested PCR and fluorescent PCR were used to amplify amelogenin and IL 3 genes from human single lymphocytes and blastomeres. Results The amplification efficiencies of nested PCR and fluorescent PCR of man single lymphocyte/woman single lymphocyte/blastomere were 64.7%/65%/85.7%, and 72%/76%/86.9% respectively. The PCR products of the lymphocyte were exactly the same as those of the donor gDNA. The PCR products of blastomeres coming from the same embryo were all identical. Conclusion The amplification efficiencies of nested PCR and fluorescent PCR are similar while fluorescent PCR is more conventional, time sparing and sensitive.