Nakanishi Hiroaki, Ohmori Takeshi, Yoneyama Katsumi, Hara Masaaki, Takada Aya, Saito Kazuyuki
Department of Forensic Medicine, Juntendo University School of Medicine, 2-1-1, Hongo, Bunkyo, Tokyo 113-8421, Japan.
National Research Institute of Police Science, 6-3-1, Kashiwanoha, Kashiwa, Chiba 277-0882, Japan.
Leg Med (Tokyo). 2020 Nov;47:101758. doi: 10.1016/j.legalmed.2020.101758. Epub 2020 Jul 15.
We investigated whether bloodstain examination and DNA typing can be performed on washed bloodstains on clothes. Blood was dropped onto T-shirts made from 100% cotton or 100% polyester. After drying, the T-shirts were hand-washed with handwashing soap, dishwashing detergent, laundry detergent, soap, or just water until the bloodstains could not be seen. After drying the T-shirts, DNA and RNA were extracted simultaneously from the bloodstained areas using commercial kits. RNA was reverse-transcribed to DNA, and then the detection of the mRNAs for HBB, ACTB, and 18S rRNA was examined. DNA was quantified via real-time PCR, and then STR typing was performed with a commercial kit. The luminol and leucomalachite green tests were used as preliminary bloodstain tests, and an immuno-chromatography kit was used to identify human bloodstains. DNA could be extracted from all washed bloodstains, but more DNA was extracted from cotton T-shirts than from polyester T-shirts. STR typing was successful for all bloodstains without issues such as PCR inhibition. In the human bloodstain identification test using mRNA, almost all bloodstains produced a Ct value for HBB and all bloodstains produced a Ct value for 18S rRNA, whereas few bloodstains produced a Ct value for ACTB. All bloodstains reacted positively to luminol, but some were negative for leucomalachite green. Most of the bloodstains did not react positively in the human bloodstain identification test using the immuno-chromatography kit. The results suggest that human bloodstain identification and DNA typing can still be performed after clothes with bloodstains are washed.
我们研究了是否可以对衣物上洗涤过的血迹进行血迹检验和DNA分型。将血液滴落在由100%棉或100%聚酯制成的T恤上。干燥后，用洗手液、洗洁精、洗衣液、肥皂或仅用水对T恤进行手洗，直到血迹看不见。T恤干燥后，使用商业试剂盒从血迹区域同时提取DNA和RNA。RNA被逆转录为DNA，然后检测HBB、ACTB和18S rRNA的mRNA。通过实时PCR对DNA进行定量，然后使用商业试剂盒进行STR分型。鲁米诺和隐色孔雀石绿试验用作初步血迹检验，免疫层析试剂盒用于鉴定人血迹。可以从所有洗涤过的血迹中提取DNA，但从棉质T恤上提取的DNA比从聚酯T恤上提取的更多。所有血迹的STR分型均成功，没有出现PCR抑制等问题。在使用mRNA的人血迹鉴定试验中，几乎所有血迹都产生了HBB的Ct值，所有血迹都产生了18S rRNA的Ct值，而很少有血迹产生ACTB的Ct值。所有血迹对鲁米诺反应呈阳性，但有些对隐色孔雀石绿呈阴性。在使用免疫层析试剂盒的人血迹鉴定试验中，大多数血迹没有呈阳性反应。结果表明，带有血迹的衣物洗涤后仍可进行人血迹鉴定和DNA分型。
