Identifying body fluids from forensic samples can provide valuable evidence for criminal investigations. Messenger RNA (mRNA)-based body fluid identification was recently developed, and highly sensitive parallel identification using reverse transcription polymerase chain reaction (RT-PCR) has been described. In this study, we developed reverse transcription loop-mediated isothermal amplification (RT-LAMP) as a simple, rapid assay for identifying three common forensic body fluids, namely blood, semen, and saliva, and evaluated its specificity and sensitivity. Hemoglobin beta (HBB), transglutaminase 4 (TGM4), and statherin (STATH) were selected as marker genes for blood, semen, and saliva, respectively. RT-LAMP could be performed in a single step including both reverse transcription and DNA amplification under an isothermal condition within 60 min, and detection could be conveniently performed via visual fluorescence. Marker-specific amplification was performed in each assay, and no cross-reaction was observed among five representative forensically relevant body fluids. The detection limits of the assays were 0.3 nL, 30 nL, and 0.3 μL for blood, semen, and saliva, respectively, and their sensitivities were comparable with those of RT-PCR. Furthermore, RT-LAMP assays were applicable to forensic casework samples. It is considered that RT-LAMP is useful for body fluid identification.

从法医样本中识别体液可以为刑事调查提供有价值的证据。最近开发了基于信使RNA（mRNA）的体液鉴定，并且已经描述了使用逆转录聚合酶链反应（RT-PCR）进行的高灵敏度平行鉴定。在这项研究中，我们开发了逆转录环介导的等温扩增（RT-LAMP）作为一种简单，快速的测定方法，用于鉴定三种常见的法医体液，即血液，精液和唾液，并评估了其特异性和敏感性。分别选择血红蛋白β（HBB），转谷氨酰胺酶4（TGM4）和斯坦汀（STATH）作为血液，精液和唾液的标记基因。RT-LAMP可以一步一步完成，包括在等温条件下60分钟内进行逆转录和DNA扩增，并且可以通过视觉荧光方便地进行检测。在每种测定中均进行了标记特异性扩增，在五个代表性的法医相关体液之间未观察到交叉反应。血液，精液和唾液的检测限分别为0.3 nL，30 nL和0.3μL，其灵敏度与RT-PCR相当。此外，RT-LAMP分析适用于法医案件样本。认为RT-LAMP对于体液识别是有用的。分别为唾液和唾液，其敏感性与RT-PCR相当。此外，RT-LAMP分析适用于法医案件样本。认为RT-LAMP对于体液识别是有用的。分别为唾液和唾液，其敏感性与RT-PCR相当。此外，RT-LAMP分析适用于法医案件样本。认为RT-LAMP对于体液识别是有用的。