Objectives: To introduce real-time polymerase chain reaction （real-time PCR） into the initial sample screening, to improve the effectiveness of traditional trace sample extraction method.
Methods: Serial diluted 9947A was quantified using a Rotor-Gene Q real-time RT-PCR, and the genotype was determined with AmpFℓSTRTM IdentifilerTM Plus PCR kit. Thus a quantitative threshold model was built to obtain complete STR typing from the trace samples. In addition, 903 trace samples were used to verify the reliability.
Results: When the samples quality concentration was >0.03 ng/μL, the effective STR typing could be directly obtained; when the concentration was >0.01 and ≤0.03 ng/μL, the effective STR typing could be directly obtained by optimizing the PCR thermal cycle parameters （30 cycles）; and when the concentration was ≤0.01 ng/μL, no effective map could be obtained even if PCR was optimized.
Conclusions: The real-time PCR quantitative threshold model is effective for the screening of trace samples.
题目: 微量生物检材定量及分型.
目的: 在传统微量生物检材提取方法中，引入荧光定量流程对样品进行初筛，以提高其有效检验。.
方法: 将DNA标准品9947A梯度稀释，使用Rotor-Gene Q实时荧光定量分析仪定量，用AmpFℓSTR™ Identifiler™ Plus PCR扩增试剂盒检测基因型，构建微量生物检材可获得完整STR分型结果的定量阈值模型，并使用903个微量检材验证其可靠性。.
结果: 检材质量浓度>0.03 ng/μL可直接获得有效的分型；检材质量浓度>0.01~0.03 ng/μL，可通过优化增加PCR热循环参数（30个循环）获得有效分型；检材质量浓度≤0.01 ng/μL时，即使优化PCR也无法得到有效图谱。.
结论: 该实时荧光定量阈值模型可有效对微量生物检材进行初筛。.
关键词: 法医遗传学；串联重复序列；实时荧光定量；微量检材.
Keywords: forensic genetics; tandem repeat sequences; real-time fluorescence quantification; trace samples.
Copyright© by the Editorial Department of Journal of Forensic Medicine.