Juusola Jane, Ballantyne Jack
Graduate Program in Biomolecular Sciences, University of Central Florida, Orlando, FL, 32816-2366, USA.
J Forensic Sci. 2007 Nov;52(6):1252-62. doi: 10.1111/j.1556-4029.2007.00550.x. Epub 2007 Sep 15.
An alternative approach to conventional protein-based body fluid identification is gene expression profiling analysis. In the present work, we report the development of sensitive and robust multiplex quantitative reverse transcriptase-PCR assays for the identification of blood, saliva, semen, and menstrual blood. Each body fluid assay comprises a triplex system that detects transcripts from two body fluid-specific genes and a housekeeping gene GAPDH. The body fluid-specific genes include erythroid delta-aminolevulinate synthase (ALAS2) and beta-spectrin (SPTB) for blood, statherin (STATH) and histatin 3 (HTN3) for saliva, protamine 1 (PRM1) and protamine 2 (PRM2) for semen, and matrix metalloproteinase 7 (MMP7) and matrix metalloproteinase 10 (MMP10) for menstrual blood. Normalization of both body fluid-specific genes to the housekeeping gene by means of appropriate cycle threshold metrics ensures the high specificity of each assay for its cognate body fluid.
与传统的基于蛋白质的体液鉴定方法不同，基因表达谱分析是另一种方法。在本研究中，我们报告了用于鉴定血液、唾液、精液和月经血的灵敏且稳健的多重定量逆转录聚合酶链反应检测方法的开发。每种体液检测方法都包含一个三重系统，该系统可检测来自两个体液特异性基因和一个管家基因甘油醛-3-磷酸脱氢酶（GAPDH）的转录本。体液特异性基因包括用于血液的红系δ-氨基乙酰丙酸合成酶（ALAS2）和β-血影蛋白（SPTB），用于唾液的富组蛋白（STATH）和组蛋白3（HTN3），用于精液的鱼精蛋白1（PRM1）和鱼精蛋白2（PRM2），以及用于月经血的基质金属蛋白酶7（MMP7）和基质金属蛋白酶10（MMP10）。通过适当的循环阈值指标将体液特异性基因与管家基因进行标准化，可确保每种检测方法对其同源体液具有高特异性。
