Influences of different RT-qPCR methods on forensic body fluid identification by microRNA
Influences of different RT-qPCR methods on forensic body fluid identification by microRNA
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不同RT-qPCR方法对microRNA法医体液鉴定的影响
DOI：
10.1016/j.fsigss.2015.09.117
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发表时间：
2015-12-01
期刊：
影响因子：
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通讯作者：
Liang, Weibo
中科院分区：
文献类型：
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作者：
Li, Zhilong;Bai, Peng;Liang, Weibo
自从Hanson试图从微阵列中寻找特异性microRNA用于体液鉴定以来，microRNA（miRNA）引起了法医学界的广泛兴趣。目前，研究组已筛选出几组可用于法医体液鉴定的miRNA标记。这些miRNA标记物中的大多数不能在不同的实验室中重复。SYBRgreen确认的标记物总是不能被Taqman的另一个小组确认。不同的逆转录定量PCR方法可能是造成这一现象的主要原因。为探讨不同RT-qPCR方法对miRNA表达的影响，本研究根据前人的研究结果，选择了3种不同RT-qPCR方法检测结果相矛盾或一致的miRNA，分别为月经血、阴道分泌物和精液中的miR-412- 3 p、miR-124 a-3 p和miR-10 b-5 p。分别采用Taqman和SYBRgreen进行分析。SYBRgreen和Taqman检测同一miRNA标记物的不同Δ Cq值。不同的结果可以归因于两种逆转录定量方法的灵敏度和特异性。应根据研究对象和样品条件选择合适的逆转录定量方法。(C)2015爱思唯尔爱尔兰有限公司版权所有。
MicroRNA (miRNA) has attracted wide interest of the forensic community since Hanson attempted to search specific microRNAs for body fluid identification from microarray. Research groups have screened out some sets of miRNA markers that can be used in forensic body fluids identification up to now. Most of those miRNA markers could not be repeated in different labs. The markers confirmed by SYBRgreen always could not be confirmed by another group in Taqman. Different reverse-transcription quantitative PCR methods might be the major cause of that. To seek out the influences of different reverse-transcription quantitative PCR methods on the expression of miRNA, we chose three miRNAs which showed conflicted or consistent results in different RT-qPCR methods according to previous studies, including miR-412-3p, miR-124a-3p, miR-10b-5p, for menstrual blood, vaginal secretions and semen respectively. MiRNAs were analyzed through Taqman and SYBRgreen according to the protocols respectively. Different Delta Cq values of the same miRNA marker were detected by SYBRgreen and Taqman. Different results can be attributed to the sensitivity and specificity of two reverse-transcription quantitative methods. The choice of appropriate reverse-transcription quantitative method should be based on the study targets and the conditions of samples. (C) 2015 Elsevier Ireland Ltd. All rights reserved.