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目的研究多重PCR-片段分析方法进行快速检测胎儿染色体异常的可行性。方法选择21、18、13、X和Y染色体分布大致均匀区域,设计38对特异性引物和通用引物共同介导的多重基因片段进行同等效率扩增,经对产物片段大小进行定量分析,使用专用软件对染色体的整倍性进行判读。采用该方法检测羊水细胞标本112例,同时与经典核型分析结果对照分析。结果建立了多重-PCR片段分析5类非整倍性染色体异常的产前诊断方法,且24小时内同时完成多个标本。112例样本进行多重PCR片段分析全部成功,52例正常女性胎儿和男性胎儿与核型分析结果完全一致;5例21-三体、4例18-三体、1例47,XXX和1例47,XYY与核型分析结果完全一致;培养失败无法进行核型分析2例,经多重-PCR片段分析排除了5类非整倍体染色体异常;多态性遗传标记35例、结构异常11例和1例嵌合体,经多重PCR-片段分析均为正常。结论建立的多重PCR-片段分析检测5类非整倍性染色体异常与传统核型分析具有符合性;并结合软件数据分析方法简便、准确、快速、通量高,可作为产前诊断5类非整倍体染色体异常的一种可行性方法。
Objective To study the feasibility of multiplex PCR-fragment analysis for rapid detection of fetal chromosomal abnormalities. Methods The chromosomes of chromosomes 21, 18, 13, X and Y were distributed in a roughly uniform region. 38 pairs of specific primers and common primers were designed to amplify the multiple gene fragments equally. After analyzing the size of the fragments quantitatively, The software interprets the euploidy of chromosomes. Using this method, 112 cases of amniotic fluid cell samples were detected, and the results were compared with the classical karyotype analysis. Results A multiplex pre-PCR method was established for the prenatal diagnosis of 5 aneuploidy chromosomal abnormalities. Multiple samples were simultaneously completed in 24 hours. The results of multiplex PCR analysis of 112 cases were all successful. The results of 52 cases of normal fetuses and fetuses were completely consistent with karyotype analysis. Five cases of 21 trisomy, 4 cases of 18-trisomy, 1 case of 47, XXX and 1 case of 47 , XYY was completely consistent with karyotyping results; 2 cases could not be karyotyped after culturing failure, and 5 kinds of aneuploidy chromosomal abnormalities were excluded by multiple-PCR fragment analysis; 35 cases were polymorphic genetic markers, 11 cases were structural abnormalities One chimera was normal by multiplex PCR-fragment analysis. Conclusions The established multiplex PCR-fragment analysis of 5 types of aneuploidy chromosomal abnormalities with the traditional karyotype analysis is consistent with; and software data analysis method is simple, accurate, fast, high throughput, can be used as prenatal diagnosis of 5 types of non- A feasible method for euploid chromosomal abnormalities.