摘要
DNATyperTM 21 试剂盒具有良好的种属特异性、分型准确性和位点扩增均衡性,可用于个体识别和亲权鉴定等法医遗传学分析.为进一步提升该试剂盒的抗抑制物能力,在存在不同浓度靛蓝、腐殖酸、血红蛋白、血红素和黑色素的情况下,探索在PCR反应体系中加入牛血清白蛋白(BSA)、牛凝血酶(BT)以及 4 种不同的DNA聚合酶:FastStart Taq DNA聚合酶(FastStart Taq)、TaKaRa Ex Taq Hot Start Version DNA聚合酶(EX Taq HS)、Klentaq DNA聚合酶(Klentaq)、MyFiTM DNA聚合酶对该试剂盒分型能力的提升程度.研究结果表明,BSA在提高STR基因座的检出率方面皆表现出高效性;BT仅有助于克服靛蓝、黑色素、腐殖酸和血红素引起的分型抑制;4 种DNA聚合酶对不同的抑制剂产生不同的抗抑制效果,其中以EX Taq HS抗抑制效果最佳,但其对血红素的抗抑制性远不如BSA和BT.本研究通过综合分析为该试剂盒后续的优化提供基础数据.
Abstract
The DNATyperTM 21 kit has good species specificity,typing accuracy,and locus amplification balance,which can be used for forensic genetic analysis such as individual identification and paternity testing.To improve the anti-inhibitor ability of this kit,the addition of bovine serum albumin(BSA),bovine thrombin(BT),FastStart Taq DNA Polymerase,TaKaRa Ex Taq Hot Start Version(Ex Taq HS),MyFiTM DNA Polymerase,and Klentaq DNA Polymerase(Klentaq)as PCR enhancers to the PCR reaction system was explored in the presence of different concentrations of inhibitors,such as indigo,humic acid(HA),hemoglobin,heme,and melanin.The results revealed that BSA exhibited high efficiency in improving the detection rate of STR loci;BT only helped to overcome the typing inhibition caused by indigo,melanin,HA,and heme;Four types of DNA polymerase had different anti-inhibitor effects on different inhibitors,among which Ex Taq HS was more effective than the other three DNA polymerases,but displayed comparatively lower resistance to hemoglobin than BSA and BT.This study provides basic data for further optimization of the kit through comprehensive analysis and facilitates its application in daily forensic investigations.
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