Department of Forensic Sciences, The George Washington University, 2100 Foxhall Road NW, Washington, DC 20007, USA; National Institute of Standards and Technology, Biomolecular Measurement Division, 100 Bureau Drive, Gaithersburg, MD 20899, USA.
Department of Forensic Sciences, The George Washington University, 2100 Foxhall Road NW, Washington, DC 20007, USA; The Bode Technology Group, 10430 Furnace Road, Suite 107, Lorton, VA 22079, USA.
Forensic Sci Int Genet. 2018 Nov;37:64-72. doi: 10.1016/j.fsigen.2018.07.019. Epub 2018 Jul 25.
The positive identification of seminal fluids in sexual assault crimes is considered crucial evidence to determine whether a sexual act occurred or not. However, current presumptive methods lack specificity and sensitivity. Confirmation of semen by microscopic examination of spermatozoa is laborious, time consuming, and can sometimes lead to negative or inconclusive results. Here we report the use of the Proximity Ligation Real-Time PCR (PLiRT-PCR) assay as an attractive and promising confirmatory method for the identification of semen and sperm proteins using two polyclonal antibodies, Prostate Specific Antigen (PSA) and Sperm-Specific Protein (SP10), respectively. PLiRT-PCR, relies on protein recognition by pairs of proximity probes (antibody-DNA conjugates) that give rise to a ligated DNA strand. The ligated DNA strand is then amplified and detected by qPCR.
在性侵犯犯罪中，精液的阳性鉴定被认为是确定是否发生性行为的关键证据。然而，目前的推定方法缺乏特异性和敏感性。通过显微镜检查精子来确认精液既费力又耗时，有时还会导致阴性或不确定的结果。在这里，我们报告了使用接近连接实时 PCR（PLiRT-PCR）检测法作为一种有吸引力和有前途的确认方法，用于使用两种多克隆抗体，即前列腺特异性抗原（PSA）和精子特异性蛋白（SP10），分别鉴定精液和精子蛋白。PLiRT-PCR 依赖于通过对接近探针（抗体-DNA 缀合物）的配对进行蛋白质识别，这些探针导致连接的 DNA 链。然后通过 qPCR 扩增和检测连接的 DNA 链。
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