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暴露于不同环境条件下的人血液、精液和唾液样本中选定 microRNAs 的稳定性。

Stability of selected microRNAs in human blood, semen and saliva samples exposed to different environmental conditions.

机构信息

Department of Cell Biology and Genetics, University of Lagos, Akoka Yaba, Lagos, Nigeria.

出版信息

Forensic Sci Int. 2022 Jul;336:111338. doi: 10.1016/j.forsciint.2022.111338. Epub 2022 May 6.

Abstract

Body fluid identification is an essential aspect of forensic investigations. MicroRNAs (miRNAs), a type of non-coding RNA has a lot of potential in forensic studies as biomarkers. Using a reference gene - GAPDH, this study ascertained the stability of hsa-mir-451a, hsa-miR-10b, and hsa-miR-205 in blood, semen and saliva respectively, exposed to different environmental conditions over different periods of time. Twenty adult males aged 20-35 donated blood, sperm, and saliva. These fluids were separated into four groups and exposed to Outdoor, Indoor, Fridge, and Freezer conditions for 37 days. Total RNA was isolated using Trizol-Isopropanol extraction protocol. FIREScript RT cDNA synthesis kit and Luna Universal qPCR Master Mix were used for cDNA synthesis and qPCR respectively. Under different environmental conditions, miR-451a and miR-10b was detected in all blood and semen samples but not in all saliva samples. Pearson's correlation test for Cq values showed multiple strong correlations (p < 0.05) within and outside the groups compared to GADPH. Regardless of the body fluid sample, the freezer group was more stable compared to other groups, although correlations are not homogeneous in all samples. Expression of the targeted genes in the body fluids showed that hsa-miR-451a, hsa-miR-10b, hsa-miR-205 could be utilized as biomarkers in forensics for body fluids identification.

摘要

体液鉴定是法医学调查的重要方面。MicroRNAs(miRNAs)作为一种非编码 RNA,在法医研究中作为生物标志物具有很大的潜力。本研究使用参考基因 GAPDH,确定 hsa-mir-451a、hsa-miR-10b 和 hsa-miR-205 在血液、精液和唾液中的稳定性,分别暴露于不同环境条件下不同时间段。20 名年龄在 20-35 岁的成年男性捐献了血液、精液和唾液。将这些液体分为四组,分别暴露于户外、室内、冰箱和冷冻条件下 37 天。使用 Trizol-Isopropanol 提取方案分离总 RNA。FIREScript RT cDNA 合成试剂盒和 Luna Universal qPCR Master Mix 分别用于 cDNA 合成和 qPCR。在不同的环境条件下,miR-451a 和 miR-10b 可在所有血液和精液样本中检测到,但并非所有唾液样本中均能检测到。与 GADPH 相比,Cq 值的 Pearson 相关检验显示组内和组间存在多个强相关性(p<0.05)。无论体液样本如何,与其他组相比,冷冻组更为稳定,尽管并非所有样本的相关性都一致。在体液中的靶基因表达表明,hsa-miR-451a、hsa-miR-10b、hsa-miR-205 可用作法医体液鉴定的生物标志物。

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