国产Goldeneye~(TM) 20A试剂盒性能指标验证

目的测试国产GoldeneyeTM20A试剂盒技术性能指标,评估其法医学应用能力。方法从方法学验证、准确性、峰值均衡性、灵敏度、批次间试剂及稳定性测试、耐受性、不同检材的适应性与一致性、种属特异性、混合样本等9个方面对GoldeneyeTM20A试剂盒进行测试。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;等位基因间的均衡性≥83%,同一荧光标记基因座间的均衡性≥55%,不同标记物间的均衡性≥52%;0.125ng DNA阳性样本可检出全部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,具有一定的种属特异性和混合DNA样本检测能力。结论国产GoldeneyeTM 20A试剂盒可用于法庭科学实际检案与建库。

The developmental validation of the homemade GoldeneyeTM 20A PCR amplification kit

Objective The technical target of the homemade GoldeneyeTM 20A kit was tested and its forensic application was estimated in this paper.Methods The validation of the GoldeneyeTM 20A kit was carried out in terms of genotyping method,precision and accuracy,peak height balance,sensitivity,stability among different kits,survivability,adaptability and consistency for various samples,species specificity and mixtures.Results The DNA genotyping results of the posotive control were accurate and the internal standard together with the ladder met the needs;Heterozygote peak height ratio imbalance≥83%,peak height ratio imbalance of the loci with the same fluorescent color≥55% and that with the different fluorescent color≥52%;STR genotyping results can all be obtained from 0.125ng DNA.Genotyping results were correct among different batches kits and after freezing/melting test.The kit could be usded in some degraded and inhibited DNA samples.DNA from various sample sources were amplified successfully using GoldeneyeTM 20A kit and consistent genotypes were obtained.The kit was highly specific for human DNA and had the ability to genotype mixtures.Conclusion All the results demonstrated that GoldeneyeTM 20A kit was suitable for criminal cases and DNA database in forensic practice.