Multiplex mRNA profiling for the identification of body fluids
Multiplex mRNA profiling for the identification of body fluids
复制标题
DOI：
10.1016/j.forsciint.2005.02.020
复制
发表时间：
2005-08-11
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
我们报告了一种多重逆转录聚合酶链反应（RT-PCR）方法的发展，用于确定法医案件分析中经常遇到的体液，即血液，唾液，精液和阴道分泌物。使用我们确定的以组织特异性方式表达的选定基因，我们开发了一种多重RT-PCR检测方法，该方法由8种体液特异性基因组成，可用于检测血液、唾液、精液和阴道分泌物的单一或混合染色。这些基因包括血液中的P-spectrin （SPTB）和porphobilinogen deaminase (PBGD)，唾液中的statherin （STATH）和组蛋白3 (HTN3)，精液中的鱼精蛋白1 （PRMI）和prolamine 2 (PRM2)，以及阴道分泌物中的人β -防御素1 （HBD-1）和粘蛋白4 （MUC4）。已知的或推测的这些基因的功能表明一个极其有限的基因表达模式，这是纳入组织特异性分析的基本要求。该方法基于基因表达谱分析，其中通过使用毛细管电泳/激光诱导荧光检测适当mRNA物种的存在来鉴定体液特异性基因。基于mrna的方法，例如本工作中描述的多重RT-PCR方法，可以方便地识别体液染色中存在的组织成分，并且可以取代目前在法医血清学实验室中使用的血清学和生化测试的电池。2005爱思唯尔爱尔兰有限公司版权所有。
We report the development of a multiplex reverse transcription-polymerase chain reaction (RT-PCR) method for the definitive identification of the body fluids that are commonly encountered in forensic casework analysis, namely blood, saliva, semen, and vaginal secretions. Using selected genes that we have identified as being expressed in a tissue-specific manner we have developed a multiplex RT-PCR assay which is composed of eight body fluid-specific genes and that is optimized for the detection of blood, saliva, semen, and vaginal secretions as single or mixed stains. The genes include P-spectrin (SPTB) and porphobilinogen deaminase (PBGD) for blood, statherin (STATH) and histatin 3 (HTN3) for saliva, protamine 1 (PRMI) and prolamine 2 (PRM2) for semen, and human beta-defensin 1 (HBD-1) and mucin 4 (MUC4) for vaginal secretions. The known or presumed functions of these genes suggest an extremely restricted pattern of gene expression, which is a basic requirement for incorporation into a tissue-specific assay. The methodology is based upon gene expression profiling analysis in which the body fluid-specific genes are identified by detecting the presence of appropriate mRNA species using capillary electrophoresis/laser induced fluorescence. An mRNA-based approach, such as the multiplex RT-PCR method described in the present work, allows for the facile identification of the tissue components present in a body fluid stain and could supplant the battery of serological and biochemical tests currently employed in the forensic serology laboratory. (c) 2005 Elsevier Ireland Ltd. All rights reserved.