荧光定量PCR技术研究线粒体DNA nt16519单核苷酸多态性

目的应用荧光定量PCR技术，建立mtDNA nt16519位点的PCR分型方法。方法应用Primer Express3．0软件，设计1对Taqman MGB探针和1埘扩增引物，在探针的5’端分别标记FAM和VIC报告荧光，应用7500荧光定量PCR和直接测序两种方法，分别榆测62份血样和18份毛发样本的mtDNA nt16519单核苷酸多态性，比较二种方法结果的一致性。结果62份血样和18份毛发样本均得到了可靠的mtDNA nt16519分型结果。结论建立的荧光定量PCR的分型方法操作简单、结果准确，适用于法医DNA检案。

mtDNA nt16519 polymorphism analysis based on real-time quantitative PCR

Objective To establish an efficient mtDNA nt16519 typing method using real-time quantitative PCR technique. Methods A pair of Taqman MGB probes labelled with fluorescent dye FAM and VIC at the 5′ end respectively, and a pair of specific primers, targeting the mtDNA nt16519, were designed using the Primer Express 3.0 software. Then, the mtDNA nt16519 polymorpism was detected using the real-time quantitative PCR and sequencing methods respectively in 80 biological samples including 62 blood samples and 18 hair shaft samples. Results All of the 80 samples were typed successfully, and all of the type results were consistent with two different methods used. Conclusion The mtDNA nt16519 polymorphism typing method based on the real-time quantitative PCR is simple and valid, and can be applied to forensic practice.