摘要
目的测试DNATyperTM15试剂盒的技术性能指标,评估其法医学应用能力。方法制定测试方案,从方法学验证、灵敏度、混合样本、批次间试剂稳定性及批量样本测试、DNA提取方法适应性测试、各类常见检材的测试、稳定性测试等8个方面进行测试,并与IdentifilerTM和PowerPlex16试剂盒进行比较。结果DNATyperTM15试剂盒灵敏度较高,批次间性能稳定,对各类案件检材和DNA提取方法具有较好的适应性,具有检验混合DNA样本检测的能力。结论DNATyperTM15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,可用于法庭科学的检案与建库。
Objective Developmental validation studies were designed according to the standards of forensic DNA community on DNATyper^TM15 kit ,which simultaneously amplifies 14 STR loci (D6S1043. D21 S11 .D7S820, CSF1PO. D2S1338. D3S1358, D13S317, D8S1179. D16S539. Penta E. D5 S818, VWA. D18S51 .FGA ) and amelogenin, a sex-determining locus. Methods Several key factors were tested including: Ⅰ. amount of hot-start Taq polymerase; Ⅱ. annealing temperature; Ⅲ sensitivity; Ⅳ. reaction volume; Ⅴ. cycle number; Ⅳ. primer concentration. DNATyper^TM15 was also compared with two other widely used commercial STR amplification kits, namely Identifiler^TM and PowerPlex. Results No difference in performance was observed with three lots of DNATyperTS15. Performance was not affected even after 20 times of repeated freeze-and-thaw. All three kits performed comparably in the following aspects: Ⅰ sensitivity ; Ⅱ. ability to genotype mixed samples ;Ⅲ amplification of DNA from various sample sources. DNA extraction methods (Chelex-100, magnetic beads, silica beads) did not result in any observable effect on performance with any of the three kits. Conclusion All the results demonstrated that DNATyper^TM 15 is suitable for both forensic DNA database work and casework.
出处
《中国法医学杂志》
CSCD
2008年第2期73-76,81,共5页
Chinese Journal of Forensic Medicine