FTA卡血样快速直接PCR扩增方法比较初探  

Preliminary study on comparison of rapid and direct PCR amplification methods with FTA blood cards

作　　者：刘琳[1,2] 项林平[3] 胡志敏[1,2] 董会 于书欣 姜伯玮[5] 赵蕾[6] 欧元[5] 赵兴春[5] 叶健[5] 

机构地区：[1]厦门市公安局,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刑警支队,重庆401320 [4]法庭科学生物芯片工程实验室,北京100038 [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》2014年第3期191-193,共3页Chinese Journal of Forensic Medicine

基　　金：国家科技支撑计划课题(2012BAK02B04-1);国家自然科学基金委员会资助项目(81202384);公安部科技强警基础工作专项(2011GABJC028)

摘　　要：目的探索FTA卡血样快速直接PCR扩增方法,比较相关方法的应用价值。方法将DNA TyperTM19试剂盒分别与3种快速试剂(Roche、TaKaRa、Fermentas)组合,以FTA卡血样为模板构建扩增体系,按照经优化的循环参数进行快速直接扩增,并与Typer19试剂盒常规方法相比较,对方法进行评价。结果 Typer19试剂盒分别与3种快速PCR检验试剂进行组合,均可实现快速直接扩增,STR分型结果均与常规方法一致,扩增用时60min,较常规扩增所需的215min缩短了约70%,但Typer19试剂盒与Roche试剂组合基因座间峰高有部分不平衡;与TaKaRa试剂组合扩增个别基因座出现双肩锋;与Fermentas试剂组合扩增峰高明显不平衡,且个别基因座存在双肩峰。结论应用DNA TyperTM19试剂盒与快速试剂组合直接PCR扩增方法,可有效缩短扩增时间,可根据效果在相关实践中选择使用。Objective Explore methods of rapid and direct PCR amplification with FTA blood cards in order to compare the application value of related methods. Methods The DNA TyperTM19 amplification kit was respectively combined with Roche, TaKaRa and Fermentas the three kinds of rapid PCR reagents. The amplification systems were established with FTA blood cards as templates. The optimized parameters were used to conduct rapid and direct amplification. The results were compared with the conventional method＇s of the TyperTM 19 kit in order to evaluate the methods. Results Rapid and direct amplification conducted by the DNA TyperTM 19 amplification kit and the three rapid PCR reagents could all obtain the STR typing results consistent with the conventional method＇s. The amplification time was 60min, which was nearly 70% ]ess than the conventional method＇s that was 215min. But there was some peak height imbalance between loci in the combination of the Typerl9 kit and the Roche reagent. Some shoulder peaks were revealed in individual loci in the combination of the Typerl9 kit and the TaKaRa reagent. There were significant peak height imbalance and shoulder peaks in individual loci in the combination of the Typerl9 kit and the Fermentas reagent. Conclusion The amplification time was significantly shortened with the direct PCR amplification conducted by the DNA TyperTM 19 amplification kit and the rapid PCR reagents. These methods could be chosen for use according to the results in relevant practice.

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

分 类 号：D919[医药卫生—法医学]