DNA Typer~(TM)15 plus直扩试剂盒在DNA数据库建设中的应用

目的测试DNA TyperTM15 plus直扩试剂盒的技术性能指标,评价其在DNA数据库建设中的应用价值。方法采用DNA TyperTM15 plus试剂盒,并使用IdentifilerTM和DNA TyperTM15试剂盒进行比较,设定不同体系和引物量、不同退火温度和循环次数以进行方法验证;设定不同模板量标准品、不同比例混合样本,取猪、狗、兔等动物的血液样品,血痕、骨骼、唾液斑等常见检材样本以及不同建库样本,以验证试剂盒灵敏度、特异性、稳定性以及混合样本、常见检材及建库样本的检测能力。结果直扩试剂盒分型结果准确,重复性好,灵敏度可达0.125ng,不同批次间试剂检测结果稳定,对不同检材有很好的适应性。10μL扩增体系时FTA卡和加强型血液采集卡取样直径应为0.5mm,而血滤纸、血液采集卡样本和经典型血液采集卡取样直径应为1.0mm。结论 DNA TyperTM15 plus直扩试剂盒的性能可以满足DNA数据库建设及检案的需要,可在相关实验中选择使用。

The application of DNA TyperTM15 plus direct kit in DNA database

Objective To test the technical performance of DNA TyperTM15 plus direct kit and evaluate its application in forensic DNA database.Methods A testing program was designed,aimed to test several key factors including method validation,sensitivity,species-specificity and stability.Mixed and different samples were also tested.The DNA TyperTM15 plus direct kit was meanwhile compared with two other widely used commercial STR amplification kits,IdentifilerTM and DNA TyperTM15.Results The DNA TyperTM15 plus direct kit got accurate and stable genotyping results,and was of good repeatability,high sensitivity(0.125ng) and strong adaptability.For the 10μL of amplification volume,the punch size of FTA card or thick card was suggested 0.5mm and the normal blood collection cards 1.0mm.Conclusions The DNA TyperTM15 plus direct kit is much suitable for both forensic DNA database work and casework,and can be selectively applied in the relevant experiments.