MicroRNA markers for forensic body fluid identification obtained from microarray screening and quantitative RT-PCR confirmation.
MicroRNA markers for forensic body fluid identification obtained from microarray screening and quantitative RT-PCR confirmation.
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DOI：
10.1007/s00414-009-0402-3
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发表时间：
2010-05
影响因子：
2.1
通讯作者：
Kayser M
中科院分区：
文献类型：
--
作者：
Zubakov D;Boersma AW;Choi Y;van Kuijk PF;Wiemer EA;Kayser M
microRNA（miRNAs）是具有重要调控功能的非蛋白质编码分子;许多具有组织特异性表达模式。原则上，它们非常小的尺寸使它们不太容易降解，不像信使RNA（mRNA），后者以前被提议作为法医体液鉴定的分子工具。为了鉴定用于法医体液鉴定的合适的miRNA标记物，我们首先使用微阵列平台筛选来自唾液、精液、阴道分泌物、静脉血和经血的总RNA样品中718种人类miRNA的表达。基于完整的基于阵列的miRNA表达谱，可以容易地将所有体液彼此区分开。来自微阵列候选标记物的定量逆转录PCR（RT-PCR; TaqMan）测定的结果证实了静脉血的几种miRNA和精液的几种其他miRNA在靶向体液中的强烈过表达。然而，没有候选标记从阵列实验的其他体液，如唾液，阴道分泌物，或经血可以通过RT-PCR确认。在实验室条件下，静脉血和精液污渍至少1年的时间降解不会显著影响所鉴定的miRNA标记物的检测灵敏度。TaqMan检测法检测选定的静脉血和精液miRNA标记物的检测限仅需要每次RT-PCR检测亚皮克量的总RNA，这大大低于可靠的mRNA RT-PCR检测通常所需的量。因此，我们提出了几个稳定的miRNA标记的法医鉴定的血迹和其他几个精液染色鉴定，使用市售的TaqMan检测的应用。在寻找其他法医学相关体液的合适miRNA标记物方面，仍有必要进行更多的工作。本文的在线版本（doi：10.1007/s00414 - 009 - 0402 - 3）包含补充材料，可供授权用户使用。
MicroRNAs (miRNAs) are non-protein coding molecules with important regulatory functions; many have tissue-specific expression patterns. Their very small size in principle makes them less prone to degradation processes, unlike messenger RNAs (mRNAs), which were previously proposed as molecular tools for forensic body fluid identification. To identify suitable miRNA markers for forensic body fluid identification, we first screened total RNA samples derived from saliva, semen, vaginal secretion, and venous and menstrual blood for the expression of 718 human miRNAs using a microarray platform. All body fluids could be easily distinguished from each other on the basis of complete array-based miRNA expression profiles. Results from quantitative reverse transcription PCR (RT-PCR; TaqMan) assays for microarray candidate markers confirmed strong over-expression in the targeting body fluid of several miRNAs for venous blood and several others for semen. However, no candidate markers from array experiments for other body fluids such as saliva, vaginal secretion, or menstrual blood could be confirmed by RT-PCR. Time-wise degradation of venous blood and semen stains for at least 1 year under lab conditions did not significantly affect the detection sensitivity of the identified miRNA markers. The detection limit of the TaqMan assays tested for selected venous blood and semen miRNA markers required only subpicogram amounts of total RNA per single RT-PCR test, which is considerably less than usually needed for reliable mRNA RT-PCR detection. We therefore propose the application of several stable miRNA markers for the forensic identification of blood stains and several others for semen stain identification, using commercially available TaqMan assays. Additional work remains necessary in search for suitable miRNA markers for other forensically relevant body fluids. The online version of this article (doi:10.1007/s00414-009-0402-3) contains supplementary material, which is available to authorized users.
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影响因子：
2.1
作者：
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影响因子：
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