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

1986 Fiscal Year Final Research Report Summary

• Project/Area Number: 60570276
• Research Category: Grant-in-Aid for General Scientific Research (C)
• Allocation Type: Single-year Grants
• Research Field: Legal medicine
• Research Institution: KOCHI MEDICAL SCHOOL
• Principal Investigator: ISHIZU HIDEO KOCHI MEDICAL SCHOOL, PROFESSOR, 医学部, 教授 (70033157)
• Co-Investigator(Kenkyū-buntansha): TSUTSUMI AKIRA KOCHI MEDICAL SCHOOL, RESEARCH ASSISTANT, 医学部, 助手 (50155420) ; YAMAMOTO YUJI KOCHI MEDICAL SCHOOL, RESEARCH ASSISTANT, 医学部, 助手 (30136379)
• Project Period (FY): 1985 – 1986
• Keywords: Specics identification / Blood stain / Body fluid stain / Monoclonal antibody / ELISA

Research Abstract

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.