Messenger RNA profiling: a prototype method to supplant conventional methods for body fluid identification
Messenger RNA profiling: a prototype method to supplant conventional methods for body fluid identification
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DOI：
10.1016/s0379-0738(03)00197-x
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发表时间：
2003-08-12
影响因子：
2.2
通讯作者：
Ballantyne, J
中科院分区：
文献类型：
--
作者：
Juusola, J;Ballantyne, J
体液识别的常规方法使用各种劳动密集型的、技术上多样化的技术，这些技术以系列而非并行的方式执行，并且在时间和样本方面是昂贵的。理论上，体液的鉴定可以通过确定在共同构成该体液的细胞中专门表达的足够数量的mRNA来进行。与传统的生化分析方法相比，基于mRNA的方法的优点包括更高的特异性，通过一个共同的分析格式，提高及时性，减少样品消耗和与DNA提取方法的兼容性，同时和半自动分析，在这份报告中，我们证明了RNA在生物染色剂中是稳定的，可以回收足够的数量和质量进行分析。使用灵敏的逆转录-聚合酶链反应（RT-PCR）检测来自管家基因S15、β-肌动蛋白和GAPDH的信使RNA在血液、精液和唾液斑中。此外，我们已经确定了一些候选组织特异性基因，statherin，histatin 3，PRB 1，PRB 2和PRB 3，可能是有用的唾液的阳性鉴定。来自这些基因的信使RNA在唾液斑中可检测到，但在血液或精液斑中却检测不到。总的来说，这些发现构成了基于原型RNA的测定系统的基础，该系统最终可能取代用于体液鉴定的常规方法。(C)2003爱思唯尔科学爱尔兰有限公司保留所有权利。
Conventional methods of body fluid identification use a variety of labor-intensive, technologically diverse techniques that are performed in a series, not parallel, manner and are costly in terms of time and sample. Theoretically, the identification of a body fluid may be made by determining a sufficient number of mRNAs that are expressed exclusively in cells that collectively comprise that body fluid. Advantages of an mRNA-based approach, compared to conventional biochemical methods of analysis, include greater specificity, simultaneous and semi-automatic analysis through a common assay format, improved timeliness, decreased sample consumption and compatibility with DNA extraction methodologies.In this report, we demonstrate that RNA is stable in biological stains and can be recovered in sufficient quantity and quality for analysis. Messenger RNA from the housekeeping genes S 15, beta-actin and GAPDH was detected in blood, semen and saliva stains using a sensitive reverse transcriptase-polymerase chain reaction assay (RT-PCR). Additionally, we have identified a number of candidate tissue-specific genes, statherin, histatin 3, PRB1, PRB2 and PRB3 that may be useful for the positive identification of saliva. Messenger RNAs from these genes were detectable in saliva stains but not in blood or semen stains. Collectively these findings constitute the basis of a prototype RNA based assay system that may eventually supplant conventional methods for body fluid identification. (C) 2003 Elsevier Science Ireland Ltd. All rights reserved.