荧光标记复合扩增检测4个STR基因座多态性

目的建立荧光标记复合扩增D1S2142,D13S1492,D14S306,D15S659基因座检测分型方法,并对成都汉族群体4个基因座的遗传多态性进行调查。方法用6-FAM标记D1S2142和D15S659引物,HEX、TMR分别标记D14S306和D13S1492引物,PCR复合扩增,310基因分析仪电泳自动收集电泳结果数据,GeneScan Analysis Software3.7NT软件计算扩增产物片段相对大小,Genotyper(3.7NT软件进行样本基因型分型,建立了荧光标记复合扩增检测4个STR基因座基因型的方法,对145名成都汉族无关个体样本进行分型。结果荧光标记复合扩增D1S2142,D13S1492,D14S306,D15S659基因座,每个STR基因座都获得了清晰的基因型分型结果。145份样本,4个STR基因座分别检出10,14,7,12个等位基因和22,54,21,39种基因型,其基因型分布均符合Hardy-W e inberg平衡。4个基因座在成都汉族群体的杂合度分别依次为0.7793,0.8345,0.7793和0.8345;多态信息含量分别依次为:0.7656,0.8730,0.7470和0.8312。累计非父排除率为0.9783,累计个人识别机率为0.9999 917。结论荧光标记复合扩增D1S2142,D13S1492,D14S306,D15S659基因座,可实现对每个基因座准确分型;成都汉族群体该4个基因座的遗传学数据,可为群体遗传学和法医学研究与应用提供基础资料。

Study on the polymorphisms of four STR loci using fluorescence - labeled multiplex - PCR technique

Objective To establish a method for typing STR loci D1S2142, D13S1492, D14S306 andD15S659 by fluorescence-labeled multiplex-PCR technique, by which the genetic data of Han population in Chengdu area were obtained. Methods Primers of STR loci D1S2142 and D15S659 were labeled with 6-FAM, and primer of D14S306 and D13S1492 were labeled with HEX and TMR respectively. Multiplex-PCR technique was applied to amplifying the target DNA. The electrophoresis of PCR products was accomplished on the 310 Genetic Analyzer with the sample's electrophoresis data collected automatically using Data Collection Software 3.0. The relative fragment lengths of PCR products were calculated by the GeneScan Analysis Software 3.7NT and genotyped by the Genotyper~ 3.7NT Software. Therefore, a method of multiplex-PCR labeled with fluorescence to detect the genotypes of the four STR loci had been established. We used this method to investigate the genotype distributions and allele frequencies in 145 EDTA-blood specimens collected from unrelated individuals in Chengdu Han population. Results Four STR loci were exactly determined by fluorescence-labeled multiplex-PCR technique. Each locus was successfully genotyped in all 145 samples. 10, 14, 7, 12 alleles and 22, 54, 21, 39 genotypes were observed in D1S2142, D13S1492, D14S306, and D15S659 locus, respectively. The distributions of genotype for four STR loci in Chengdu Han population were in accordance with Hardy-Weinberg equilibrium. The heterozygotes and polymorphism information component (PIC) of the four STR loci were 0.7793, 0.8345, 0.7793, 0.8345 and 0.7656, 0.8730, 0.7470, 0.8712 respectively. The combined exclusion probability and the combined discrimination power of the four STR loci in Chengdu Han population were 0.9783 and 0.9999917. Conclusion The STR loci D1S2142, D13S1492, D14S306, and D15S659 could be correctly genotyped using fluorescence-labeled multiplex-PCR technique. The population genetic information of the four STR loci may be used as the basic data in the study and application in genetics and forensic practice.