多重荧光错配引物PCR进行单核苷酸多态性检测分析

Single nucleotide polymorphisms analysis using multiplex fluorescence mismatching PCR

目的发展一种快速、检测位点数适中(5～20)、既适用于中小规模科研也适用于临床检测的单核苷酸多态性(SNPs)分型方法。方法通过改进等位基因特异性聚合酶链反应(PCR),加入公用荧光引物,发展一种利用毛细管电泳的多重荧光错配引物PCR方法,对已知基因型的8份健康者样本的5个线粒体SNPs位点进行分型。结果一条公用荧光引物即可同时完成5个SNPs位点的分型,其结果与已知基因型完全一致。结论多重荧光错配引物PCR在中小规模科研及临床检测中具有经济、快速的优势,该方法尤其适用于微量DNA样本的检测。

Objective To develop a new method for multiple typing of single nucleotide polymorphisms（SNPs） suitable for clinical detection.Methods This method was developed from allele-specific polymerase chain reaction（ASPCR） by introducing universal fluorescent primers to construct a new multiplex fluorescence mismatching PCR utilizing capillary electrophoresis.Samples from eight healthy subjects were detected for five mitochondria SNPs types by newly constructed method.Results Five SNPs could be typed simultaneously by using one universal fluorescent primer,and the detected results was fully consistent with known genotypes.Conclusion Multiplex fluorescence mismatching PCR could be economic and rapid for small and medium-sized scientific research and clinical detection,which might be especially suitable for detecting minor DNA samples.