23-plex Y-SNPs复合扩增体系在法医学应用的研究  

Development of 23-plex Y-SNPs Typing Method Based on Single-base Extension and Capillary Electrophoresis Technology and Its Application in Forensic Science

作　　者：张爱平[1,2,3] 刘超[1,2] 李越[1] 刘长晖[1] 陈丽伟[1,2] 葛璐璐[1] 陈晓晖[1] 

机构地区：[1]广州市公安局刑事科学技术研究所,广东广州510030 [2]中山大学中山医学院法医学系,广东广州510080 [3]佛山市顺德区公安局,广东佛山528300

出　　处：《中山大学学报（医学科学版）》2010年第5期685-691,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家十一五科技支撑计划课题(2006BAK07B01);公安部重点科技攻关项目(20034421401)

摘　　要：【目的】应用SNaPshot技术建立23-plexY-SNPs的复合扩增体系,研究其在法医学中的应用价值。【方法】从Y染色体单体群命名树上选择23个Y-SNP位点:P164、P203、P148、P145、M89、P151、M216、P128、P157、P149、P131、P199、P123、P191、P201、M111、M9、P132、P200、P197、M119、P136和M134,进行复合PCR扩增,PCR产物纯化后利用荧光标记单碱基延伸技术(SNaPshot试剂盒)结合毛细管电泳技术对单核苷酸多态性进行检测,应用该体系调查了290名广东地区无亲缘关系的男性个体。【结果】所建立的23个Y-SNPs检测体系,各位点均具有遗传多态性,基因多样性范围为0.0137～0.4912。在290名男性中检测到143个单体型,单体型多样性为0.9907。【结论】该23个Y-SNPs检测体系能快速而准确的对样本进行SNP分型,在法医学、人类学和相关学科的研究中具有较高的应用价值。Objective To develope a robust single nucleotide polymorphism（SNPs） typing assay with co-amplification and to type 23 human Y chromosome SNPs using SNaPshot kit.Methods Markers were drawn from the phylogenetic tree of Y chromosome：P164,P203,P148,P145,M89,P151,M216,P128,P157,P149,P131,P199,P123,P191,P201,M111,M9,P132,P200,P197,M119,P136,and M134.All 23 Y-SNPs in one reaction were amplified;the pooled PCR products were purified.The minisequencing reactions were performed simultaneously for all 23 Y-SNPs with fluorescein-labeled dideoxynucleotides.A total of 290 male Chinese in Guangdong were genotyped by capillary electrophoresis and multicolour fluorescence detection.Results A 23-plex Y-SNPs co-amplification system was developed.All of the Y-SNPs are polymorphic in Chinese.The gene diversity of the Y-SNPs ranged from 0.013 7-0.491 2.A total of 143 haplotypes were found,the haplotype diversity was calculated to be 0.990 7.Conclusion The 23-plex Y-SNPs system developed is efficient,high-throughput and appears to be suitable for forensic science and anthroponomy.

关 键 词：个体识别 单碱基延伸(微测序) SNaPshot试剂盒 单核苷酸多态性 Y染色体 

分 类 号：R325[医药卫生—人体解剖和组织胚胎学]