19种扩增试剂盒的确证试验  

Validation of 19 Kits for PCR Amplification

作　　者：张庆霞 刘小芳 路志勇 王顺霞 

机构地区：[1]北京市公安司法鉴定中心,北京100192 [2]北京市昌平区公安司法鉴定中心,北京102200

出　　处：《刑事技术》2016年第6期507-511,共5页Forensic Science and Technology

摘　　要：目的测试11种国产试剂盒(包含DNATyperTM15、Goldeneye16A、Goldeneye16BT、Goldeneye16C、Goldeneye20A、Goldeneye BASIC、AGCU17+1、AGCU Expressmarker16、AGCU Expressmarker22、AGCU Expressmarker20、STRtyper-21G)和8种进口试剂盒(包含Identifiler、ID-Direct、ID-Plus、Sinofiler、Power Plex16、Power PTheselex16HS、Power Plex18D、Power Plex21)的技术性能指标,评估其法医学应用能力,并对各种试剂盒进行分析比较。方法制定测试方案,从方法学、准确性、均衡性、灵敏度、批次间试剂的稳定性、耐受性、适应性与一致性、种属特异性、混合样本、不同反应体系的适应性等10个方面进行测试。结果阳性DNA样本分型正确,内标和等位基因分型标准物符合要求;等位基因间、同一荧光标记基因座间及不同标记物间的均衡性较好;试剂盒使用说明书中推荐的最小DNA模板量的阳性DNA样本均能检出全部STR基因座分型;不同批次间和反复冻融后试剂盒测试可以获得正确分型;对降解检材和混有抑制剂的样本等具有一定的耐受性;能对案件中多种检材进行分型且分型结果一致;各种试剂盒均具有种属特异性和混合DNA样本检测的能力;各种试剂盒在扩增体系为25μL、10μL时均可以得到完整的DNA分型。结论所检测试剂盒在上述性能指标等方面已经达到国际同类产品的技术水平,可用于法庭科学的实际检案与建库。Objective The performance of 11 homemade PCR amplification kits （DNATyperTmlS, Goldeneyel6A, Goldeneyel6BT, Goldeneyel6C, Goldeneye20A, Goldeneye BASIC, AGCU17＋I, AGCU Expressmarkerl6, AGCU Expressmarker22, AGCU Expressmarker20, STRtyper-21G） was to test and their forensic applicability evaluated together with the 8 foreign counterparts （Identifiler, ID-Direct, ID-Plus, Sinofiler, PowerPlexl6, PowerPlexl6HS, PowerPlexl8D, PowerPlex2 l） in this paper. Methods Validation protocols were designed to comply with the guidelines i：~sued by the Scientific Working Group on DNA Analysis Methods （SWGDM）. The evaluation of the 19 kits for PCR amplification was carried out in terms of genotyping method, precision and accuracy, peak height balance, sensitivity, stability, survivability, adaptability and consistency for various samples, species specificity, mixed samples, and the suitability into different reaction systems. Results The DNA genotyping results from all the 19 kits were accurate with the positive control sample while the requirements were met by their internal standard and ladder. The 19 kits for PCR amplification were able to be amplified successfully and to genotype clearly and efficiently, meanwhile there was no peak height imbalances leveling at high sensitivity obtained. Genotyping results were correct among different batches of the same kits even subjected to repeated freezing/melting. The 19 kits were also capable of being used in some degraded and/or inhibitory DNA samples, and well performed with revealing the consistent genotypes for the DNA from various sources. These kits were highly specific for human DNA and able to genotype for mixed samples. For different amplifying reaction setting-ups, the 19 kits were capable of getting the consistent genotypes. Conclusion All the 19 kits for PCR amplification are suitable for both the routine casework and DNA database construction in forensic practice.

关 键 词：法医物证学 扩增试剂盒 确证试验 短串联重复序列 

分 类 号：D919.4[医药卫生—法医学]