新鲜血痕样本直接扩增试验条件比较

Experimental condition of fresh bloodstains using direct PCR amplification kits

目的探讨新鲜血痕在不同直接扩增试剂盒的试验条件。方法存放2d内的新鲜血痕FTA卡540份和存放1～3m的陈旧性血痕FTA卡270份，各分别随机均分为3组，每份血痕打3片1．0mm纸片，分别用ONATyper^TM15Plus试剂盒、Goldeye^TM20A试剂盒和华夏。”试剂盒在标准条件（说明书条件）、优化条件1（标准条件＋1μLDMSO）和优化条件2（标准条件＋室温浸泡1h）扩增检验，比较在3种条件下各组STR检验成功率。结果陈旧血痕样本用3种直扩试剂盒，在3种条件下，检验成功率均〉97．00％，且均高于新鲜血痕。新鲜血痕在标准条件和优化条件1下，成功率为27．22％～31．67％，在优化条件2下，检验成功率相似（〉97．00％），与标准条件和优化条件1比较，有统计学差异（P〈0．01）。结论新鲜血痕在加入直接扩增试剂后于室温浸泡1h，可有效提高STR检验成功率。

Objective To investigate the experimental conditions of fresh bloodstains using three different direct PCR amplification kits. Methods 540 fresh bloodstains （ ≤2d） and 270 aged bloodstains （ 1 -3m） were randomly divided into 3 groups, respectively. Three pieces with 1.0 mm were obtained from eachcard, which was amplified with DNATyperTM15 Plus, GoldeyeTM20A, and HuaxiaTM direct amplification kits according to standard conditions （ manufacturer instructions ）, optimum conditions 1 （ standard conditions ＋ 1μL DMSO）, and optimum conditions 2（standard conditions ＋ immersed lh at room temperature）, respectively. Results The STR successful rate of aged bloodstains was more than 97.00% under the three conditions using different direct amplification kits, respectively （P 〉 0. 05）, which was higher than that of fresh bloodstains. The successful rate of fresh bloodstains ranged from 27.22% to 31.67% according to standard and optimum conditions 1 （ P 〉 0.05 ）. Under optimum conditions 2, the successful rates of fresh bloodstains were consistent with those of aged bloodstains （ 〉 97.00% ）. There were significant differences between optimum conditions 2 and standard or optimum conditions 1 （P 〈 0. O1 ）. Conclusions We suggested that the successful rates of ~~esh bloodstains could significantly improve when direct amplification kits were added and immersed 1 h at room temperature.