快速PCR扩增体系的建立及法医学应用  

The establishment and application in forensic science of rapid PCR amplification

作　　者：刘琳[1,2] 项林平[3] 胡志敏[1,2] 董军磊[4] 姜伯玮[5] 白雪[6] 赵蕾[6] 欧元[6] 赵兴春[6] 叶健[6] 

机构地区：[1]厦门市公安局思明分局,福建厦门361003 [2]中国人民公安大学,北京100038 [3]重庆市公安局巴南区分局,重庆401320 [4]天津市公安局西青分局,天津300380 [5]公安部第一研究所,北京100048 [6]公安部物证鉴定中心,北京100038

出　　处：《中国法医学杂志》2015年第1期32-35,共4页Chinese Journal of Forensic Medicine

基　　金：国家科技支撑计划课题(2012BAK02B04-1);国家自然科学基金资助项目(81202384);公共安全生物物证检验系列试剂研制与示范应用(Z131100004513007)

摘　　要：目的建立快速PCR扩增体系,探讨其在法医实践中的应用价值。方法设置不同的缓冲液、酶浓度与Mg SO4浓度、热循环参数等,观察不同参数对扩增结果的影响,建立快速PCR扩增体系。使用该体系对常见检材进行STR分型,与常规方法扩增的结果比较,考察其分型准确性与扩增用时。结果本文建立的快速PCR扩增体系含DNA TyperTM19试剂盒的Primer Mix(5×)2μL;Ta KaRa试剂的Fast BufferⅡ(10×)1μL,Speed STARTMHS DNA Polymerase(5U/μL)0.3μL,Mg SO4(50mmol/L)为0.1μL。扩增程序为95℃120s;95℃2s、61℃15s,27个循环;72℃60s。结论快速PCR扩增体系分型结果准确,用时仅19min,缩短了常规扩增用时,有较大的法医学应用价值。Objective To establish the rapid PCR amplification system anti evaluate its appnc~ forensic practice. Methods By adopting various buffers, enzyme concentrations, MgSO4 molarities and adjusting thermal cycle parameters, this paper closely observed the influence incited by the above variation, thus establishing a rapid PCR amplification system, with which the paper STR typed some common materials and compared the results obtained by using it and conventional method so as to determine its accuracy and time consumed. Results The rapid PCR amplification was composed of 2μL Primer Mix （ 5 × ） of DNA TyperTM19 amplification kit, IμL Fast Buffer Ⅱ （10 × ） of TaKaRa rapid PCR test reagent, 0.3μL SpeedSTARTM HS DNA Polymerase （ 5U/μL ） and 0. 1μL MgSOn（50mmol/L） with a process made up of one 120s reaction of 95℃, 27 cycle reactions of one 2s of 95℃ and one 15s of 61℃, as well as a 60s one of 72℃. Conclusion This rapid PCR amplification system could obtain accurate STR typing results. Besides, time required was 19min. The time of conventional amplification was greatly reduced. It is practical and promising in forensic practice.

关 键 词：法医物证学 DNA TyperTM19试剂盒 STR 快速PCR 

分 类 号：D919[医药卫生—法医学]