Gc亚型检测的复合MS-PCR法及其应用

Study of Gc-subtype genotyping by multiplexed MS-PCR and application

目的探讨建立Gc亚型检测的复合MS-PCR法及其应用价值。方法根据Gc基因中的2处点突变,设计2对片段相差5bp的等位基因特异性引物和1条公共引物进行复合MS-PCR,分析Gc多态性,并调查武汉地区218例汉族无关个体Gc多态性和鉴定10例亲子关系。结果复合MS-PCR检测的Gc基因型,与AmpliTypePM试剂盒的分型结果一致;武汉地区汉族人群Gc基因的3个常见等位基因Gc1F、Gc1S、Gc2的基因频率分别为0.4816、0.2592、0.2592,观察杂合度(Hobs)、期望杂合度(Hexp)、多态性信息含量(PIC)、个人识别能力(DP)、非父排除率(PE)分别为0.6193、0.6359、0.6253、0.7974、0.3480,基因型分布符合Hardy-Weinberg平衡;真三联体和非真三联体亲子鉴定各5例,前者不排除父子关系,与常规STR分型一致,后者经Gc-MS-PCR分型排除2例。结论建立复合MS-PCR法检测Gc亚型在法医物证鉴定中有实用价值。

Objective To establish a new multiplexed MS-PCR method for Gc-sub typing and study the application. Methods Gc was genotyped by multiplexed MS-PCR involving two allele-specific primers differed by 5bp length and a common reverse primer corresponding to two point mutations.218 unrelated individuals of Han in Wuhan and 10 parentage testing cases were genotyped using multiplexed MS-PCR. Results Typing results by multiplexed MS-PCR were distinct and consistent with those by Ampli Type PM PCR. The allele frequency of Gc*1F 、Gc*1S and Gc*2 were 0.48、0.26、0.26 respectively. The observed heterozygosity (Hobs)、 the expected heterozygosity (Hexp)、the polymorphism information content (PIC)、 the discrimination power (DP) and the probability of exclusion (PE) were 0.6193、0.6359、 0.6253 、0.7974 and 0.3480 respectively. The population data fitted the Hardy-Weinberg law. The results of Gc genotyped by multiplexed MS-PCR of 5 true-trios were consistent with STR analysis. 2 cases were excluded by Gc-MS-PCR in 5 false-trios. Conclusion multiplexed MS-PCR method is reliable and useful for forensic identification.