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.

血迹的物种鉴定在法医医学实践中占有重要地位，为此，已经报告了各种免疫学方法。最近，具有高特异性和可重现性的许多针对人血蛋白的单克隆抗体已在商业上获得。在目前的工作中，我们已经建立了使用针对人白蛋白（HALB）和免疫球蛋白kappa链（HIGK）的单克隆抗体鉴定人类血迹的ELISA方法。 ELISA对HALB的敏感性极高，该方法可以检测到1 ng的纯化HALB。通过这种ELISA方法，可以从人类血液中以稀释量从人体血液中检测到HALB，并从人类血迹（1天大）提取物中以稀释（稀释为稀释）（对血迹的原始血液体积稀释）。在4年零12年的旧血迹中，HALB在摘录中被证明了多达200万次和十万次稀释。在Halb的Elisa中，除了tamarin以外的灵长类动物外，没有观察到与非人类动物的交叉反应。稀释一十万次。可以从人类的血迹（1天大）提取物中证明希格克，并分别从1000倍的稀释液和1年和4岁的血迹的提取物中，分别有五千次和十万次稀释。 ELISA对HIGK的物种特异性优于ELISA HALB的ELISA，只有黑猩猩的血液显示出交叉反应。这些结果表明，使用对HALB和HIGK的单克隆抗体的当前ELISA方法可用于鉴定人类血液和人类血液和人类血液和人类血液和希格。法医学领域的血迹。