DNATyper Y21直扩试剂盒技术指标测试及应用价值评价

The technical index test and experimental validation of DNATyper Y 21 PCR amplification kit

目的测试DNATyper Y21直扩试剂盒的技术性能指标,评估其在法医学中的应用价值。方法收集270份无关男性血液采集卡、80份常见检材和40份日常检案样本。对DNATyper Y21直扩试剂盒方法学(扩增体系、引物量、退火温度和循环次数),技术指标(灵敏度、稳定性、种属特异性,对混合样本、常见检材及建库样本的检测能力)进行测试。结果该试剂盒分型准确、图谱清晰,各等位基因峰高均>400相对荧光单位,最佳条件为10μL体系,1×引物量,59℃退火温度,28次循环。试剂盒准确性、一致性、均衡性、种属特异性均良好,最小检出量为0.125ng,女性DNA存在下(300∶1)仍然具有高灵敏性,且在不同批次试剂间结果稳定,对骨骼、血痕、唾液斑等不同常见检材有较好的适应性。结论 DNATyper Y21直扩试剂盒可应用于法庭科学实际检案与建库。

Objective This research was aimed to test and evaluate the technical performance of DNATyper Y21 direct amplification kit. Methods 270 unrelated male blood collection cards, 80 common forensic samples and 40 routine casework samples were collected for validation of the DNATyper Y21 direct amplification kit , which was carried out in terms of methodology （amplification reaction system, primers dosage, annealing temperature and cycle numbers）, technical indicators （sensitivity, species-specificity, detection capability for mixed samples, routine samples and those used for forensic DNA database ）. Results Accurate and clear genotyping results were obtained and the peak height for each allele was over 400 relative fluorescence units. The optimum reaction was performed in 10p, L reaction volume added 1 x primer amount, and was amplified for 28 cycles with 59℃ annealing temperature. The results were of good accuracy, consistency, peak height balance, species-specificity as well as high sensitivity. Genotypes of 21 loci could be obtained from 0. 125ng genomic DNA and mixed with sample female/male DNA （300 ： 1 ）. In addition to good stability among different batches of reagents, the kit was highly adaptive to various samples including bones, blood stains and saliva spots. Conclusion The DNATyper Y 21 direct amplification kit is much suitable for database in forensic both criminal cases and DNA practice.