多重定量荧光PCR在常见染色体非整倍体疾病快速产前诊断中的应用研究

Application of Multiplex Quantitative Fluorescence Polymerase Chain Reaction (QF-PCR) for the Rapid Prenatal Diagnosis of Common Chromosome Anueploidies

目的:建立适合于中国人群特点的常见染色体非整倍体疾病快速产前诊断技术。方法:随机抽取50例正常无关个体的外周静脉血和9例羊水标本以及3例血清学筛查为阳性的胎儿羊水,应用定量荧光PCR技术分析13、18、21、X/Y染色体上的短串联重复序列(STR)的多态信息含量(PIC)。结果:中国人13、18、21、X/Y染色体上STR:D13S631、D13S634、D13S742、D18S386、D18S391、D18S535、D21S11、D21S1270、IFNAR、X22、XHPRT的PIC分别为0.800、0.749、0.821、0.914、0.623、0.760、0.876、0.848、0.848和0.807。应用以上STR对血清学筛查阳性的胎儿羊水DNA标本进行诊断,证实了18-三体综合征、21-三体综合征和Klinefelter综合征胎儿各1例,与染色体核型分析结果相一致。结论:本文研究所选STR位标在中国人群中呈现较高的遗传多态性,符合Hardy-Weinberg平衡定律,建立了快速产前诊断常见染色体非整倍体疾病的多重定量荧光PCR技术。

Objective: To apply the technique of multiplex quantitative fluorescence polymerase chain reac- tion (QF-PCR) for the rapid prenatal diagnosis of common chromosome anueploidies in Chinese population. Methods: We chose the high polymorphic short tandom repeat (STR) markers in chromosomes of 13, 18, 21, X/Y, in which aneuploidies occurred more often than others. DNA was extracted from 50 unrelated normal people, 9-amniotic fluid DNA samples for QF-PCR to establish the technique for the rapid prenatal diagnosis. Results: D13S631, D13S634,D13S742,D18S386,D18S391,D18S535,D21S11,D21S1270,IFNAR,X22,XHPRTwere analyzed by QF-PCR, whose PIC were 0.800,0.749,0.821,0.914,0.623,0.760,0.876,0.848,0.848,0.807 respectively. In addition, we diagnosed three abnormal amniotic fluid sanyles of aneuploidies, 18 trisomy, 21 trisomy and Klinefelter Syndrome, by serum screening. The results were in accordance with karyotype. Conclusion: The result of our study showed that the STR markers we chose were all highly polymorphic and suitable for prenatal diagnosis of chromosome anueploidies in accordance with Hardy-Weinberg.