A SENSITIVE METHOD FOR SPECIES IDENTIFICATION OF BLOOD AND BODY FLUID STAINS USING MONOCLONAL ANTIBODIES

使用单克隆抗体对血液和体液染色进行灵敏物种鉴定的方法

• 批准号: 60570276
• 负责人: ISHIZU HIDEO
• 金额: $ 0.96万
• 依托单位: KOCHI MEDICAL SCHOOL
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-60570276/
• 关键词: Specics identification; Blood stain; Body fluid stain; Monoclonal antibody; ELISA

Species identification of bloodstains occupies an important place in the practice of forensic medicine, and various immunological methods have been reported for this purpose. Recently, a number of monoclonal antibodies against human blood proteins with high specificity and reproducibility have become available commercially. In the present work, we have established ELISA methods for the identification of human bloodstains using monoclonal antibodies against human albumin (HALB) and immunoglobulin kappa chain (HIgk). The sensitivity of the ELISA for HALB was extremely high and 1 ng of purified HALB could be detected by this method. By this ELISA method, HALB could be detected from human blood with ten million times dilution and from a human bloodstain (1 day old) extract with two million times dilution (dilution against the original blood volume of the bloodstain). In old bloodstains of 4 years and 12 years, HALB was demonstrated in the extracts up to two million times and one hundred thousand times dilutions, respectively. In the ELISA for HALB, no crossreaction was observed with nonhuman animals except for the primates other than tamarin.In the ELISA for HIgk, the lowest detectable amount of purified human IgG was 0.1 <mu> g and human blood could be identified up to one hundred thousand times dilution. HIgk could be demonstrated from a human bloodstain (1 day old) extract with one hundred thousand times dilution and from extracts of 1 year and 4 years old bloodstains with fifty thousand times and ten thousand times dilutions, respectively. The species specificity of the ELISA for HIgk was superior to the ELISA for HALB, and only chimpanzee blood showed a cross-reaction.These results indicate that the present ELISA methods using monoclonal antibodies against HALB and HIgk are useful for the identification of human blood and bloodstains in the field of forensic medicine.

血痕的种属鉴定在法医学实践中占有重要地位，各种免疫学方法已被报道用于此目的。近年来，许多具有高特异性和可重复性的抗人血蛋白的单克隆抗体已经商业化。本研究建立了用抗人白蛋白（HALB）和免疫球蛋白κ链（HIgk）单克隆抗体鉴定人血痕的ELISA方法。该方法灵敏度极高，可检出1ng的HALB。通过这种ELISA方法，HALB可以从稀释1000万倍的人血液和从稀释200万倍（相对于血迹的原始血液体积稀释）的人血迹（1天）提取物中检测。在4年和12年的陈旧血迹中，HALB分别在200万倍和10万倍稀释液中显示。在HALB的ELISA中，除绢毛猴外，与非人动物无交叉反应，在HIgk的ELISA中，纯化的人IgG的最低检测量为<mu>0.1g，人血可鉴定至10万倍稀释。HIgk可从稀释10万倍的人血痕（1日龄）提取物和稀释5万倍和1万倍的1年和4年龄血痕提取物中证实。HIgk的种属特异性优于HALB的种属特异性上级，且仅与黑猩猩血有交叉反应，表明本文建立的HALB和HIgk单克隆抗体ELISA方法可用于法医学领域对人血和血痕的鉴定。