Screening and confirmation of microRNA markers for distinguishing between menstrual and peripheral blood
Screening and confirmation of microRNA markers for distinguishing between menstrual and peripheral blood
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用于区分经血和外周血的 microRNA 标记物的筛选和确认
DOI：
10.1016/j.fsigen.2017.05.012
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发表时间：
2017-09-01
影响因子：
3.1
通讯作者：
Zhang, Lin
中科院分区：
文献类型：
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作者：
Li, Zhilong;Bai, Peng;Zhang, Lin
犯罪现场残留的经血（MB）和外周血（PB）的识别对于犯罪重建至关重要，特别是在性侵犯中。 MicroRNA (miRNA) 是一类非蛋白质编码分子，已被证明是法医案件中体液鉴定的可行工具。几个小组已经寻找了不同体液特有的 miRNA。血液已得到最广泛的研究。然而，仅五项研究涉及经血，并且证实特定 miRNA 存在的结果无法在其他研究中重现。在本研究中，我们尝试使用 Exiqon 的 miRCURY (TM) LNA Array 筛选能够区分经血和外周血的新标记物。根据微阵列结果选择了5个miRNA，即miR-141-3p、miR-373-3p、miR-497-5p、miR-143-5p和miR-136-5p，其表达水平与对照组相比分别表现出27.95、17.96、16.74、10.14和9.21倍的变化。外周血中的水平。在确认步骤中应用TaqMan水解探针（简称TaqMan）和SYBR Green荧光染料（简称SYBR Green）两种经典的定量方法，研究不同定量方法对结果的影响。 TaqMan 确认了三种 miRNA（miR-141-3p、miR-497-5p 和 miR-143-5p），SYBR Green 确认了一种 miRNA（miR-141-3p）。此外，应用生物信息学方法来解释候选miRNA。我们的结果建立了体液鉴定的多步骤程序，并表明当使用 miRNA 鉴定血液样本的来源时，定量方法的选择很重要。 (C) 2017 Elsevier B.V. 保留所有权利。
The identification of menstrual blood (MB) and peripheral blood (PB) left at a crime scene is crucial for crime reconstruction, especially in sexual assaults. MicroRNAs (miRNAs), a class of non-protein coding molecules, have been demonstrated to be a viable tool for body fluid identification in forensic casework. Several groups have searched for miRNAs that are specific to different body fluids. Blood has been studied the most extensively. However, menstrual blood was only involved in five studies, and the results confirming the presence of specific miRNAs could not be reproduced in other studies. In this study, we attempted to screen new markers that can differentiate between menstrual blood and peripheral blood by using Exiqon's miRCURY (TM) LNA Array. Five miRNAs were selected based on the microarray results, namely, miR-141-3p, miR-373-3p, miR-497-5p, miR-143-5p, and miR-136-5p, whose expression levels exhibited 27.95-, 17.96-, 16.74-, 10.14-, and 9.21-fold changes, respectively, compared to the level in peripheral blood. Two classic quantitative methods, TaqMan hydrolysis probes (TaqMan for short) and SYBR Green fluorochrome (SYBR Green for short), were applied in the confirmation step to study the impact of different quantitative methods on the results. Three miRNAs (miR-141-3p, miR-497-5p, and miR-143-5p) were confirmed by TaqMan and one (miR-141-3p) by SYBR Green. Furthermore, bioinformatic methods were applied to interpret the candidate miRNAs. Our results established a multi-step procedure for body fluid identification and showed that the choice of quantitative method is important when miRNAs are used to identify the origin of blood samples. (C) 2017 Elsevier B.V. All rights reserved.