Haas C, Klesser B, Maake C, Bär W, Kratzer A
Institute of Legal Medicine, Forensic Genetics, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
Forensic Sci Int Genet. 2009 Mar;3(2):80-8. doi: 10.1016/j.fsigen.2008.11.003. Epub 2008 Dec 25.
mRNA profiling is a promising new method for the identification of body fluids from biological stains. Major advantages of mRNA profiling are the possibility of detecting several body fluids in one multiplex reaction and of simultaneously isolating DNA without loss of material. A reverse transcription endpoint polymerase chain reaction (PCR) method and a realtime PCR assay were established for the identification of blood, saliva, semen, vaginal secretions and menstrual blood, and were compared to conventional enzymatic and immunologic tests. The results for specificity, sensitivity and suitability to biological stains were satisfying and mRNA stability was demonstrated for up to 2-year-old stains. Two novel multiplex assays were created with the endpoint PCR primers: multiplex 1 amplifies two markers for each of the above mentioned body fluids and is suited for screening; multiplex 2 was designed for the detection of blood, vaginal secretions and menstrual blood. The results demonstrate that both endpoint PCR and realtime PCR are suitable for the identification of body fluids in forensic stains and represent an effective alternative to conventional enzymatic and immunologic tests.
mRNA分析是一种用于从生物污渍中识别体液的很有前景的新方法。mRNA分析的主要优点是有可能在一次多重反应中检测多种体液，并能在不损失材料的情况下同时分离DNA。建立了逆转录终点聚合酶链反应（PCR）方法和实时PCR检测方法，用于识别血液、唾液、精液、阴道分泌物和月经血，并与传统的酶法和免疫法检测进行比较。特异性、敏感性以及对生物污渍的适用性结果令人满意，并且证明了长达两年的污渍中mRNA的稳定性。用终点PCR引物创建了两种新型多重检测方法：多重检测1针对上述每种体液扩增两个标记，适用于筛查；多重检测2设计用于检测血液、阴道分泌物和月经血。结果表明，终点PCR和实时PCR都适用于法医污渍中体液的识别，是传统酶法和免疫法检测的有效替代方法。
