多重PCR联合DHPLC诊断21三体综合征方法的建立

目的初步建立多重PCR联合DHPLC技术检测21-三体综合征的方法。方法针对21号染色体上特异的3个STR位点(D21S1435、D21S1411、D21S11)设计引物,对100例外周血标本(其中25例21三体标本)提取基因组DNA进行多重PCR扩增,得到的PCR产物通过DHPLC仪进行结果分析。结果 DHPLC分析100例标本中21例为21-三体标本,其中20例21-三体标本的诊断结果与染色体核型分析结果一致,DHPLC技术检测21-三体异常灵敏度和特异度分别为80%,98.67%。结论 DHPLC技术诊断外周血标本结果与染色体核型诊断结果具有同一性,DHPLC技术可以对21三体综合症作出快速、较为准确的诊断。

Development of multiplex PCR-DHPLC diagnosing method for Trisomy 21

Objective: To study the rapid diagnosis of trisomy 21 by Multiplex PCR-DHPLC(Denaturing high performance liquid chromatography).Method:Specific primers were designed and synthesized according to the 3 STR sites(D21S1435,D21S11 and D21S1411) on chromosome 21,DNA samples were extracted from peripheral blood of 100 cases(25 cases were trisomy 21 samples) and amplified using Multiplex PCR,and then the amplification products were analyzed by Denaturing high performance liquid chromatography.Results: Multiplex PCR-DHPLC showed that 21 out of 100 cases were identified as trisomy 21,and 20 cases were coincident with chromosome karyotype analysis.The sensitivity and specificity of Multiplex PCR-DHPLC in the detection of trisomy 21 were 80% and 98.67%.Conclusion: The diagnosis using Multiplex PCR-DHPLC or chromosome karyotype analysis show high concordance,and Multiplex PCR-DHPLC is a reliable method in rapid diagnosis of trisomy 21.