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目的：建立一种对性连锁遗传病胎儿进行产前性别诊断的方法及探讨利用孕妇外周血中胎儿细胞检测ＺＦＹ基因的可行性。方法：用计算机辅助设计的两对寡核苷酸引物位于染色体ＺＦＹ基因保守区内，并建立了巢式聚合酶链反应（ｎｅｓｔｅｄＰＣＲ）技术扩增男性特异性的ＺＦＹ基因。结果：扩增产物分别为３５４ｂｐ和３０７ｂｐ，对不同标本的扩增结果表明，该引物具有高度的敏感性和特异性，２０份男性标本中均发现特异扩增片段，但在２０份女性标本中均未发现。应用该引物对３１份孕妇外周血连续监测，胎儿ＺＦＹ基因检出率随孕周增加而增高，总准确率达９６８％（３０／３１），４５份绒毛标本的阴性／阳性之比为１：１１４３，接近实际的胎儿性别比。结论：用巢式ＰＣＲ技术检测ＺＦＹ基因具有特异、敏感、快速、准确的优点，提示该法检测胎儿细胞ＤＮＡ是可行的，在临床上可用于性连锁遗传病的产前性别诊断。
OBJECTIVE: To establish a method for prenatal diagnosis of sexually transmitted diseases in fetuses and to explore the feasibility of using the fetal cells in pregnant women to detect ZFY gene. METHODS: Two pairs of oligonucleotide primers designed by computer were located in the conserved region of ZFY gene and a nested-PCR technique was established to amplify the male-specific ZFY gene. Results: The amplified products were 354bp and 307bp, respectively. The amplification results of different samples showed that the primers were highly sensitive and specific. Specific amplified fragments were found in 20 male samples. However, in 20 female samples Have not found. Using this primer, the peripheral blood of 31 pregnant women was continuously monitored. The detection rate of fetus ZFY gene increased with the increase of gestational weeks, with a total accuracy of 968% (30/31). The negative / positive ratio of 45 samples was 1: 1 143, close to the actual fetal sex ratio. Conclusion: The detection of ZFY gene by nested PCR has the advantages of specificity, sensitivity, speed and accuracy. It suggests that this method is feasible for the detection of fetal cell DNA and is clinically useful for prenatal diagnosis of sex-linked genetic diseases.