6种STR试剂盒法医学检验性能比较OACHSSCD
Comparison of the Forensic Testing Performance of Six STR Kits
目的 比较6种常用STR试剂盒的技术性能指标,评估其法医学检验能力.方法 分别采用AmpFℓSTRTM IdentifilerTM Plus PCR扩增试剂盒(简称IdentifilerTM Plus)、AGCU EX28CS荧光检测试剂盒(简称AGCU EX28CS)、NHID® 25A 案件试剂盒(液体,简称 NHID® 25A)、VerifilerTM Plus PCR 扩增试剂盒(简称VerifilerTM Plus)、VersaPlexTM 27PY System(简称 VersaPlexTM 27PY)和 FastDirectTM DNA身份鉴定系统25A(简称FastDirectTM 25A)检测不同质量法医DNA标准品9948的STR基因座的分型情况,评估其在检测灵敏度、等位基因插入/缺失、等位基因峰高、杂合子均衡性上的差异.在复合扩增体系中分别加入不同浓度的腐殖酸、氯化血红素、靛蓝、无水氯化钙和乙二胺四乙酸(ethylenediaminetetraacetic acid,EDTA)等PCR抑制剂,比较 6 种试剂盒对 PCR 抑制剂的最大耐受浓度.结果 IdentifilerTM Plus、AGCU EX28CS 和VersaPlexTM 27PY的灵敏度低至100 pg,FastDirectTM 25A、NHID® 25A和VerifilerTM Plus的灵敏度低至50 pg.不同模板质量下,5种试剂盒的STR基因座检出数均多于IdentifilerTM Plus.对50 pg及以下模板检测时,IdentifilerTM Plus的等位基因插入/缺失出现总次数低于其余5种试剂盒.AGCU EX28CS在200和100 pg模板质量下、VersaPlexTM 27PY在50 pg模板质量下检测出的等位基因峰高明显低于IdentifilerTM Plus(P＜0.05).在抗抑制性方面,NHID® 25A对腐殖酸(≤250 ng/μL)、氯化血红素(≤0.6 mmol/L)、无水氯化钙(≤1.4 mmol/L)的 抗 抑 制 性 强 于 其 他 试 剂 盒,VersaPlexTM 27PY对靛蓝(≤0.048 mol/L)的抗抑制性最强,FastDirectTM 25A 对 EDTA(≤1.0 mmol/L)的抗抑制性最强.对于氯化血红素,AGCU EX28CS 和FastDirectTM 25A的抗抑制性低于IdentifilerTM Plus,另3种试剂盒高于IdentifilerTM Plus;对于其他抑制剂,5种试剂盒的抗抑制性均达到甚至高于IdentifilerTM Plus.结论 与IdentifilerTM Plus相比,AGCU EX28CS、FastDirectTM 25A、NHID® 25A、VersaPlexTM 27PY和VerifilerTM Plus在检测灵敏度、等位基因插入/缺失、等位基因峰高、杂合子均衡性和抗抑制性总体上无明显劣势,在部分技术指标上甚至更优.
Objective To compare the technical performance indicators of six commonly used STR kits,and to evaluate their capabilities in forensic testing.Methods The STR genotyping results of fo-rensic DNA standard sample 9948 with different template quantities were analyzed using IdentifilerTM Plus,AGCU EX28CS,NHID® 25A,VerifilerTM Plus,VersaPlexTM 27PY,and FastDirectTM 25A.Differen-ces in detection sensitivity,allelic drop-out/in,allele peak height and heterozygote balance were eva-luated.Various PCR inhibitors,including humic acid,heme,indigo,calcium ions and ethylenediamine-tetraacetic acid(EDTA),were added to the multiplex amplification systems to compare the maximum tolerance levels of the six kits to PCR inhibitors.Results The detection sensitivity of IdentifilerTM Plus,AGCU EX28CS and VersaPlexTM 27PY was as low as 100 pg,whereas the sensitivity of FastDi-rectTM 25A,NHID® 25A and VerifilerTM Plus reached 50 pg.At different template quantities,the num-ber of STR loci detected by the five kits was greater than that of IdentifierTM Plus.When detecting templates of 50 pg or less,the total number of allelic drop-out/in observed with the IdentifilerTM Plus was lower than that observed with the other five kits.The allele peak heights detected by AGCU EX28CS at 200 and 100 pg template quantities and by VersaPlexTM 27 PY at 50 pg were significantly lower than those detected by IdentifilerTM Plus(P＜0.05).Regarding resistance to inhibitors,NHID® 25A showed stronger tolerance to humic acid(≤250 ng/μL),heme(≤0.6 mmol/L)and calcium ions(≤1.4 mmol/L)than other kits.It was found that VersaPlexTM 27PY showed the strongest tolerance to indigo(≤0.048 mol/L);and FastDirectTM 25A showed the strongest tolerance to EDTA(≤1.0 mmol/L).For heme,AGCU EX28CS and FastDirectTM 25A showed lower tolerance than IdentifilerTM Plus,whereas the other three kits exhibited higher tolerance.For other inhibitors,all five kits demonstrated tolerance comparable to or higher than that of IdentifilerTM Plus.Conclusion Compared with IdentifilerTM Plus,AGCU EX28CS,FastDirectTM 25A,NHID® 25A,VersaPlexTM 27PY and VerifilerTM Plus show no ob-vious overall disadvantage in detection sensitivity,allelic drop-out/in,allele peak height,heterozygote balance and inhibitor resistance and even demonstrate superior performance in certain technical indica-tors.
孙溢华;陈剑;李永生
苏州市公安局吴中分局物证鉴定室,江苏 苏州 215104苏州市公安局吴中分局物证鉴定室,江苏 苏州 215104苏州市公安局吴中分局物证鉴定室,江苏 苏州 215104
医药卫生
法医遗传学短串联重复序列灵敏度杂合子均衡性PCR抑制剂试剂盒
forensic geneticsshort tandem repeatsensitivityheterozygote balancePCR inhibitortesting kits
《法医学杂志》 2026 (1)
26-33,8
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