Juusola Jane, Ballantyne Jack
Graduate Program in Biomolecular Science, University of Central Florida, PO Box 162366, Orlando, FL 32816-2366, USA.
Forensic Sci Int. 2003 Aug 12;135(2):85-96. doi: 10.1016/s0379-0738(03)00197-x.
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 S15, 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.
传统的体液识别方法使用多种劳动密集型、技术多样的技术，这些技术以系列而非并行的方式进行，在时间和样本方面成本高昂。理论上，通过确定足够数量仅在共同构成该体液的细胞中表达的mRNA，可以进行体液识别。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、通过通用检测形式进行同时半自动分析、提高及时性、减少样本消耗以及与DNA提取方法的兼容性。在本报告中，我们证明RNA在生物污渍中稳定，并且可以以足够的数量和质量回收用于分析。使用灵敏的逆转录聚合酶链反应检测法（RT-PCR）在血液、精液和唾液污渍中检测到管家基因S15、β-肌动蛋白和甘油醛-3-磷酸脱氢酶的信使RNA。此外，我们已经鉴定出一些候选的组织特异性基因，即statherin、组蛋白3、PRB1、PRB2和PRB3，它们可能有助于唾液的阳性识别。这些基因的信使RNA在唾液污渍中可检测到，但在血液或精液污渍中未检测到。这些发现共同构成了基于RNA的检测系统原型的基础，该系统最终可能取代传统的体液识别方法。
