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法医学杂志 2016, Vol. 32 Issue (2) :109-113    DOI: 10.3969/j.issn.1004-5619.2016.02.008
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六色荧光标记快速PCR扩增体系的建立及验证
刘亚举1,张俊涛2,金海英3,石美森4
(1. 许昌市公安局刑事科学技术研究所,河南 许昌 461000; 2. 襄城县公安局刑侦大队,河南 襄城 461700; 3. 宁波海尔施基因科技有限公司,浙江 宁波 315000; 4. 中国政法大学 证据科学教育部重点实验室,北京 100088)
Establishment and Verification of 6-color Fluorescent-labeled Rapid PCR Amplification System
LIU YA-JU1, ZHANG JUN-TAO2, JIN HAI-YING3, SHI MEI-SEN4
(1. Institute of Criminal Science and Technology, Xuchang Public Security Bureau, Xuchang 461000, China; 2. Criminal Investigation Team, Xiangcheng Public Security Bureau, Xiangcheng 461700, China; 3. HEALTH Gene Technologies Incorporation, Ningbo 315000, China; 4. Key Laboratory of Evidence Science, Ministry of Education, China University of Political Science and Law, Beijing 100088, China)

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摘要 目的 建立PCR快速扩增程序和体系,并对其技术指标进行验证。 方法 采用六色荧光标记技术,对24个常染色体STR基因座、1个Y-STR基因座和Amelogenin、Y-InDel基因座进行复合扩增及毛细管电泳检测,同时考察体系的灵敏度、特异性、同一性、稳定性、混合样本及批量样本测试,并观察各种常见检材及降解、脱落细胞检材的分型情况。 结果 所建立的体系灵敏度达0.062 5 ng,快速扩增仅耗时65 min就可获得准确分型;种属特异性高;各种纸质血样本和混合、降解、脱落细胞检材的分型正确。 结论 本研究建立的快速扩增体系可显著提升检验速率,分型准确、稳定,对建立STR数据库、研究群体遗传学和进行法医学鉴定有重要意义。
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刘亚举
张俊涛
金海英
关键词法医遗传学   短串联重复序列   快速PCR   复合STR扩增试剂     
Abstract: Objective To establish the rapid PCR amplification program and system and to verify the technical indexes. Methods PCR multiplex and capillary electrophoresis detection of 24 autosomal STR loci and one Y-STR loci using the 6-color fluorescence marking technology, as well as Amelogenin and Y-InDel. Meanwhile, sensitivity, specificity, identity, stability, mixing and a batch of sample tests were investigated, and the genotype of various routine samples and degraded, exfoliated cell samples were observed. Results The sensitivity of the system was 0.062 5 ng. In addition, the genotype could be detected accurately only around 65 min via rapid amplification. The species-specificity was high and the genotyping of all kinds of dry blood specimens of filter paper and mixed, degraded, exfoliated cell samples were accurate. Conclusion The rapid amplification system can significantly improve the detection rate, and obtain accurate and stable genotyping results, which may be important implications for the establishment of STR database and study on population genetics and forensic identification.
Keywordsforensic genetics   short tandem repeat   rapid PCR   multiplex STR detection     
Corresponding Authors: 石美森,女,博士,教授,硕士研究生导师,主要从事法医物证学和群体遗传学研究;E-mail:shimeisen2000@163.com   
About author: 刘亚举(1978—),男,副主任法医师,主要从事法医DNA检验和群体遗传学研究;E-mail:horse3697@126.com
引用本文:   
刘亚举, 张俊涛, 金海英, 等.六色荧光标记快速PCR扩增体系的建立及验证[J]  法医学杂志, 2016,32(2): 109-113
LIU YA-JU, ZHANG JUN-TAO, JIN HAI-YING, ET AL..Establishment and Verification of 6-color Fluorescent-labeled Rapid PCR Amplification System[J]  , 2016,32(2): 109-113
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