A New Method for Determination of ABO Blood Groups using Nitrocellulose

Research Project

Project/Area Number	61870031
Research Category	Grant-in-Aid for Developmental Scientific Research
Allocation Type	Single-year Grants
Research Field	Legal medicine
Research Institution	Osaka University
Principal Investigator	SHIKATA Ichiro Osaka University Medical School (Professor), 医学部, 教授 (10035371)
Co-Investigator(Kenkyū-buntansha)	FUJITANI Noburu Osaka University Medical School (Assistant), 医学部, 助手 (10156888)
Project Period (FY)	1986 – 1987
Project Status	Completed (Fiscal Year 1988)
Budget Amount *help	¥3,200,000 (Direct Cost: ¥3,200,000) Fiscal Year 1987: ¥800,000 (Direct Cost: ¥800,000) Fiscal Year 1986: ¥2,400,000 (Direct Cost: ¥2,400,000)
Keywords	Nirocellulose / ABO-blood-group system / ABC-method / Fingerprint / 血痕
Research Abstract	(1) ABO grouping was carried out on fingerprints lifted onto nitrocellulose membrane using the ABC method or mixed coll agglutination reaction. Specimens of fingerprints were obtained by gentle pressing of fingertips onto nitrocellulose membrenes by lifting the fingerprints experimentally prepared on glass slides onto nitrocellulose sembrenes .1:Mixed cell agglutination reaction was performed by incubating the membranes with anti-A, anti-B serum or anti-HUlec lectin followed by washing and further incubation with A.B or O type erythrocytes. This technic is relatively simpler and clearer rather than the conventional method using sticking film plate and is very useful in the practice of blood lyping of fingerprints in the forensic science field. 2:Above specimens were sensitized with anti-A and anti-B sera os varying dilutions. The specifically absorbed antibodies were visualized by the ABC method. In result. group A and AB fingerprints showed positive staning for anti-A whercas group B … More and O fingerprints ahowed no positive staining. in a similar manner, group B and AB fingerprints showed positive staining for anti-B. whereas group A and O fingerprints showed no positive staining. Any positive staining with appreciable internsity could not be observed in this experiment. (2) We evaluated the advantage of using nitrocellulose beads as immunoadsorbents for ABH blood groups. The blood group substances in hemolyzed and highly diluted blood specimens were absorbed on the boads. after which the blood type was determined either by mixed cell agglutination or by the absorption-elusion technique. The latter was found to be more sensitive than the former. An optimal absorption of the ABH substances was obtained when we incubated 1 mg nitrocellulose beads with 10 ml of the blood samples for 1 hour at 25゜C. A 100000-fold diluted antigen of the A or B type or a 50000-fold diluted antigen of the H type was detected under these conditions. No procedures of extraction. purification. or concentrations of the antigen were needel. It is suggested that the new methods described above will find wider application in the practice of medico-legal examination of trace evidence. Less