单细胞单轮二重PCR诊断X-连锁鱼鳞病

Diagnosing X-linked Ichthyosis by Monoplast Single-round Duplex PCR

X-连锁鱼鳞病(X-linked ichthyosis,XLI)是隐性遗传病,是人类最常见的先天性代谢疾病之一.从单细胞水平诊断XLI,就可为开展XLI的胚胎植入前遗传学诊断(preimplantation genetic diagnosis,PGD)打下基础.采用单轮二重PCR扩增患者和正常女性的单淋巴细胞以及正常人单卵裂球的STS基因和amelogenin(Amel)基因,在正常人单淋巴细胞组和正常人单卵裂球组中STS基因扩增成功率分别为95.8%、90.9%,在患者单淋巴细胞组中的假阳性率为1.5%;在Amel-X和Amel-Y基因位点扩增成功率分别为91.9%、92.7%,污染率分别为1.4%、0;ADO的发生率为6.2%.结果表明单细胞单轮二重PCR诊断XLI具有较高的准确性和特异性,有助于我们进一步开展XLI的PGD.

X-linked ichthyosis （XLI） is a sort of sex linked recessive inherited diseases. It is one of the most common congenital metabolic diseases of the mankind. Diagnosing XLI on monoplast level using monoplast PCR, that can provide a basis for preimplantation genetic diagnosis（PGD） of XLI. The STS gene and the amelogenin （A reel） gene were amplified by single-round duplex PCR with single lymphocytes from the patient, the normal female and single blastomere from the normal person. The amplification success rates were 95.8% and 90.9% on STS gene in the groups of single lymphocytes and single blastomere from the normal person, respectively. The false positive rate was 1.5% on STS gene in the groups of single lymphocytes from the patient. The amplification success rates were 91.9% and 92.7% on the gene loci of Amel-X and Amel-Y, respectively. The contamination rates were 1.4% and 0 on the above 2 gene loci, respectively. The allele dropout rate （ADO） was 6.2% on the Amel loci. To some extent, the protocol of monoblast single-round duplex PCR for diagnosing XLI is accurate, specific and reliable. It will help us further develop the PGD of XLI.