【摘要】 种植前遗传学诊断（ｐｒｅｉｍｐｌａｎｔａｔｉｏｎ ｇｅｎｅｔｉｃｄｉａｇｎｏｓｉｓ，ＰＧＤ） 又称胚胎种植前遗传学诊断，是基于体外受精（ｉｎ ｖｉｔｒｏｆｅｒｔｉｌｉｚａｔｉｏｎ，ＩＶＦ）和早期胚胎活检（ｅａｒｌｙｅｍｂｒｙｏ ｂｉｏｐｓｙ）技术基础之上的一种新的遗传病产前诊断的方法。假肥大型肌营养不良症（ＤＭＤ／ＢＭＤ）作为一种常见的Ｘ－ 连琐隐性致死性遗传病，是国际上ＰＧＤ界最早选择进入ＰＧＤ 临床应用的几种遗传病之一。本文报道我们所建立的ＤＭＤ基因和ＳＲＹ 基因（（ｓｅｘ － ｄｅｔｅｒｍｉｎａｔｉｏｎ ｒｅｇｉｏｎ Ｙ，ＳＲＹ） 单淋巴细胞三重（ＤＭＤ） 基因外显子１９，４８ 及ＳＲＹ基因）及四重（ＤＭＤ基因外显子１７ 、４４ 、４５ 及ＳＲＹ 基因） 套式ＰＣＲ技术及其可靠性分析。采取外周血标本制备单淋巴细胞悬液，应用上述三重或四重套式ＰＣＲ 分析５０ 个正常男性单淋巴细胞，结果显示ＤＭＤ 基因外显子１７ 、１９、４４ 、４５、４８ 及ＳＲＹ 基因单淋巴细胞ＰＣＲ 扩增成功率分别为９２ ％ 、９６ ％ 、９４％ 、９６％ 、９４ ％ 以及９４－ ９６％ 。进一步分析５０ 个正常女性以及５０ 个ＤＭＤ 男性患者（缺失ＤＭＤ 基因外显子４４ ?更多还原

【Abstract】 Preimplantation genetic diagnosis (PGD), a new prenatal diagnosis which can be offered to the couples who are at risk to give births to genetically affected offspring,is based on the techniques of in vitro fertilization (IVF) and early embryo biopsy.As one of the most common X?linked lethal genetic diseases,Duchenne muscular dystrophy(DMD) is one of the first group of genetic diseases which were chosen to be evaluated the possibility of clinical PGD.Here we present that the development of 3?multiplex(including DMD gene exon19,48 and SRY gene)and 4?multiplex(including DMD gene exon17 44,45and SRY gene)PCR amplifying both DMD gene and SRY gene on single lymphocytes and the analysis of their reliability.After analysis of 50 single normal male lymphocytes with these techniques,we found the successful amplification rates on single lymphocytes for DND gene exon17,19,44,45,48,and SRY gene are 92%,96%,94%,96%,94%and 94?96%respectively.We also found that the total amplification accurate rate for 3?multiplex and 4?multiplex are 96%(607/630)and 96%(807/840) respectively along with total false positive amplification rate 4.4%(4/90)and 3.3%(4/120) respectively after further analysis of 50 normal female lymphocytes and 50 DMD patient lymphocytes carrying a DMD gene deletion mutation involved exon44?49. Our results suggested that these techniques can be used for clinical PGD application which can analyze both sex and deletion mutation of DMD gene of the embryos at risk of being affected in DMD families carrying DMD gene deletion mutations involved exon17,19,44,45 and 48.These techniques are better than those only based on either sexdetermination of DMD deletion analysis which are available so far.更多还原