Song L, Liu S, Wu H, Fang S P, F Y F
Department of Criminal Science and Technology, Xihu District Branch of Hangzhou Public Security Bureau, Hangzhou 310013, China.
Zhejiang Key Laboratory of Forensic Science and Technology, Institute of Forensic Science, Zhejiang Province Public Security Bureau, Hangzhou 310009, China.
Fa Yi Xue Za Zhi. 2018 Jun;34(6):656-658. doi: 10.12116/j.issn.1004-5619.2018.06.017. Epub 2018 Dec 25.
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.
Serial diluted 9947A was quantified using a Rotor-Gene Q real-time RT-PCR, and the genotype was determined with AmpFℓSTR Identifiler 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.
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.
The real-time PCR quantitative threshold model is effective for the screening of trace samples.
将实时聚合酶链反应（real-time PCR）引入初始样本筛查，以提高传统微量样本提取方法的有效性。
使用Rotor-Gene Q实时逆转录聚合酶链反应对系列稀释的9947A进行定量，并使用AmpFℓSTR Identifiler Plus PCR试剂盒确定基因型。由此建立定量阈值模型，以从微量样本中获得完整的STR分型。此外，使用903份微量样本验证可靠性。
当样本质量浓度>0.03 ng/μL时，可直接获得有效的STR分型；当浓度>0.01且≤0.03 ng/μL时，通过优化PCR热循环参数（30个循环）可直接获得有效的STR分型；而当浓度≤0.01 ng/μL时，即使优化PCR也无法获得有效的图谱。
实时PCR定量阈值模型对微量样本筛查有效。
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