PuriTyper~(TM)法医学纯化试剂盒的性能验证

目的验证PuriTyperTM纯化试剂盒各项性能指标和法医学应用价值。方法收集及制备抗凝血液、常见案件检材(唾液、烟头、精液、毛发、指甲、骨骼及组织块)、斑痕样本(血斑、唾液斑、精斑)以及模拟添加抑制剂和模仿自然环境中放置的血斑。采用PuriTyperTM纯化试剂盒提取纯化并进行DNA定量,IdentifilerTM复合扩增试剂盒扩增,产物经ABI 3130遗传分析仪进行检测,Genemapper软件分析结果,对该试剂盒灵敏度、稳定性、重复性、检材适应性进行测试。结果采用该试剂盒提取0.1～40μL血液分别获得0.042～26.45ng/μL的DNA。3种斑痕样本DNA产量平行试验结果稳定。不同类型检材重复检验所获IPC的CT平均值在27.60至28.03之间。常见案件检材所得分型与已知结果均一致。结论 PuriTyperTM纯化试剂盒能够满足法医DNA检验的要求,对法医学实践具有重要的应用价值。

Validation of the PuriTyperTM forensic DNA purification kit

Objective To verify the performance indexes and application value of the PuriTyperTM forensic DNA purification kit in forensic science.Methods Samples were collected and prapared,including anticoagulated blood,case sameples(saliva,butt,semen,hair,fingernail,bone and organization block),stain samples(bloodstain,salivary stain and seminal stain),and bloodstains placeed in imitated outdoor environment or in which inhibitors were added.DNA was extracted and purified with the PuriTyperTM forensic DNA purification kit,and then quantified.After ampification with the IdentifilerTM kit,the PCR products were separated on ABI3130 genetic analyzer and then analyzed by Genemapper software.The sensitivity,stability,repeatability and testing adaptability of the PuriTyperTM kit were verified.Results The PuriTyperTM forensic DNA purification kit owned the merits of high sensitivity(0.042ng/μL to 26.45ng/μL for 0.1～40 μL of blood),good repeatability(the stable DNA yields in parallel tests of 3 kind of stain samples) and stability(the average IPC CT values between 27.75 and 28.20).The typing results of selected case samples were consistent with the known control.Conclusion The PuriTyperTM forensic DNA purification kit can be applied in forensic DNA testing.