多重置换扩增在植入前胚胎性别诊断中的应用

Application of MDA in sex diagnosis of preimplanted embryos

目的用多重置换扩增(MDA)方法,对植入前胚胎的单个卵裂球全基因组扩增后进行植入前胚胎性别的遗传学诊断。方法用单细胞多重置换扩增方法对20个胚胎共20个单卵裂球进行全基因组扩增,用荧光PCR检测SRY、AMEL、XHPRT及X22 4个性别相关的基因座,检测胚胎性别,同时用同一胚胎另一卵裂球FISH结果作为对照。结果 20个卵裂球MDA均扩增成功,扩增成功率100%。20个胚胎均得到诊断,性别诊断成功率100%,其中12例为男性胚胎,8例为女性胚胎,与FISH结果一致率100%,诊断准确率100%。结论可以用单卵裂球MDA后荧光PCR检测多个基因座的方法对胚胎性别进行快速、准确的植入前诊断。

【Objective】 To determine the gender of preimplanted embryos using multiple displacement amplification（MDA） protocol of single blastomere.【Methods】 We removed 20 single blastomeres from 20 embryos,and proceeded whole genomic amplification using MDA,then detected SRY,AMEL,XHPRT,X22 makers using fluorescent PCR to determine sex.Blastomeres from the same embryo were proceeded fluorescent in situ hybridization（FISH） at the same time as positive control.【Results】 All the 20 blastomeres were successful amplified and 20 of them achieved gender determination outcome,12 of them were male,and 8 were female.Compared with FISH,the accuracy rate of fluorescent PCR was 100%.【Conclusion】 This is an efficient and accurate protocol for preimplantation gender determination using SRY,XHPRT,AMEL and X22 makers by fluorescent PCR afer multiple displacement amplification.