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不同环境条件下血斑样本中 miRNA 的稳定性和持久性。

The persistence and stability of miRNA in bloodstained samples under different environmental conditions.

机构信息

Department of Forensic Medicine, Chongqing Medical University, Chongqing 400010, China.

Hainan Medical University, Hainan 570000, China.

出版信息

Forensic Sci Int. 2021 Jan;318:110594. doi: 10.1016/j.forsciint.2020.110594. Epub 2020 Nov 14.

Abstract

miRNA markers have been an area of forensic interest to identify body fluid sources in recent years. In this study, reverse transcription and quantitative real time polymerase chain reaction (RT-qPCR) were performed to detect the existence of blood-specific miRNA markers in bloodstained samples under different environmental conditions, Blood samples from 6 individuals were deposited onto glass plates and exposed to different temperature, humidity, ultraviolet light intensity, and natural condition. When samples were stored to a series of estimated test times, total RNA was extracted and the Ct values of the target RNAs were detected, targets included two miRNA markers (hsa-miR-16-5p, hsa-miR-451a) and one reference gene (U6 snRNA). Analysis results showed that miR-451a represented strong stability and could be detected at all detection points. Meanwhile, each RNAs exhibited unique degradation characteristics, compared to U6, miRNAs showed stronger stability. Additionally, rain had an adverse effect on RNAs stability and accelerates its degradation rate.

摘要

近年来,miRNA 标志物已成为法医领域鉴定体液来源的一个研究热点。本研究通过逆转录和实时定量聚合酶链反应(RT-qPCR)检测了在不同环境条件下血斑样本中血液特异性 miRNA 标志物的存在。将 6 名个体的血液样本滴在玻璃载玻片上,并分别暴露于不同的温度、湿度、紫外光强度和自然条件下。当样本被储存至一系列估计的检测时间时,提取总 RNA 并检测目标 RNA 的 Ct 值,目标包括两个 miRNA 标志物(hsa-miR-16-5p、hsa-miR-451a)和一个参考基因(U6 snRNA)。分析结果表明,miR-451a 表现出很强的稳定性,可在所有检测点检测到。同时,与 U6 相比,每种 RNA 均表现出独特的降解特征,miRNAs 表现出更强的稳定性。此外,雨水对 RNA 稳定性有不利影响,会加速其降解速度。

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