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目的建立24个基因座的复合扩增系统,并对其性能指标进行评价。方法选择24个基因座(包含D8S1179、D5S818、D2S1338、D18S51、D6S1043、D2S441、D3S1358、vWA、D19S433、D16S539、CSF1PO、Penta D、D22S1045、D13S317、D1S1656、D7S820、TPOX、Penta E、D10S1248、TH01、D12S391、D21S11、FGA等23个STR基因座及1个Amelogenin基因座);合成引物,上游端标记荧光染料;收集DNA数据库建库血痕及口腔拭子样本及相关11种动物样本,采用本文方法进行复合扩增;并对方法的准确性、灵敏度、稳定性、种属特异性、检材适用性、混合样本的检验等系统性能指标进行验证。结果本文复合扩增系统对最长保存9年的各种血痕检材完整分型成功率为100%,且均衡性良好,无非特异性扩增,口腔拭子的成功率为97.8%,灵敏度达125pg,对含有抑制剂样本,在血红素浓度≤600μmol/L,腐殖酸浓度≤50ng/μL时检出效果稳定,种属特异性好。结论本文24个基因座复合扩增系统在DNA数据库建设中具有较好的应用价值。
Objective To establish a multiplex amplification system of 24 loci and to evaluate its performance. Methods Twenty-four loci (including D8S1179, D5S818, D2S1338, D18S51, D6S1043, D2S441, D3S1358, vWA, D19S433, D16S539, CSF1PO, PentaD, D22S1045, D13S317, D1S1656, D7S820, TPOX, PentaE, D10S1248, TH01, 23 STR loci such as D12S391, D21S11 and FGA, and 1 Amelogenin locus); synthetic primer, upstream fluorescent dye; collection of DNA database database of blood stains and oral swab samples and related animal samples of 11, using the method of this article Compound amplification; and verify the system performance indicators such as accuracy, sensitivity, stability, species specificity, sample applicability and mixed sample test. Results The successful rate of complete typing of various bloodstains samples stored for up to 9 years was 100% with a good balance and non-specific amplification. The success rate of buccal swab was 97.8%, and the sensitivity was 125pg, containing inhibitors samples, the heme detection efficiency ≤ 600μmol / L, humic acid concentration ≤ 50ng / μL stable, species-specific good. Conclusion The 24 loci multiplex amplification system in this paper has a good application value in the construction of DNA database.