Yamamoto Y, Tsutsumi A, Ishizu H
Department of Legal Medicine, Kochi Medical School, Japan.
Forensic Sci Int. 1989 Jan;40(1):85-95. doi: 10.1016/0379-0738(89)90171-0.
A series of experiments was conducted to establish a species identification method by means of the enzyme-linked immunosorbent assay (ELISA) using monoclonal antibody with high specificity, the result of which is reported. The limit of human IgG detection by inhibition ELISA using mouse anti-human immunoglobulin kappa light chain (Ig kappa) monoclonal antibody is 0.01 microgram. Using this method it was possible to detect human Ig kappa from bloodstains in dilutions up to 1:100,000 (dilution multiple of bloodstain extract to the original amount of blood). Human Ig kappa was detectable in human bloodstains in dilutions up to 1:50,000 one year after stain preparation and in dilutions up to 1:10,000 4 years after stain preparation, showing that this method is applicable to species identification of old bloodstains. When primates other than human were studied for specificity, red-chested tamarin and Japanese monkey showed no cross reaction at all. Cross reaction was noted with a chimpanzee, but experimentally it could be differentiated from human. Furthermore, no cross reaction was noted with respect to blood samples from non-primates (guinea pig, sheep, horse, dog and cat).
进行了一系列实验，以建立一种使用具有高特异性的单克隆抗体通过酶联免疫吸附测定（ELISA）进行物种鉴定的方法，并报告了实验结果。使用小鼠抗人免疫球蛋白κ轻链（Igκ）单克隆抗体的抑制ELISA检测人IgG的极限为0.01微克。使用这种方法，可以从稀释倍数高达1:100,000（血迹提取物相对于原始血量的稀释倍数）的血迹中检测到人Igκ。在血迹制备一年后，在稀释倍数高达1:50,000的人血迹中可检测到人Igκ，在血迹制备4年后，在稀释倍数高达1:10,000的人血迹中可检测到人Igκ，这表明该方法适用于陈旧血迹的物种鉴定。当研究除人类以外的灵长类动物的特异性时，红胸狨猴和日本猕猴完全没有交叉反应。与黑猩猩有交叉反应，但通过实验可以将其与人类区分开来。此外，对于来自非灵长类动物（豚鼠、绵羊、马、狗和猫）的血样没有观察到交叉反应。
