Immunochemical studies on blood group A substance from human hair

Blood group A-active substance was extracted from urea-treated human hair uith methanol-ethyl ether 1:1, v/v) or chloroform-methanol (1:1, v/v). The serological activity of blood group A substance in the hair was destroyed by A-decomposing enzyme from Clostridium tertium with concomitant development of blood group H activity. It is concluded therefore that the extract from the hair of group A contained blood group A-active glycolipid with N-acetylgalactosamine as the non-reducing sugar.     

An unusual variant of blood group A

A blood specimen from a forensic science case appeared to violate Landsteiner's Rule. The red cells failed to react with anti-A, anti-B, or O serum while reacting strongly with Ulex europaeus lectin but not other anti-A lectins. The saliva from the person involved was found to contain both A and H blood group substances in a ratio of 4:1. The blood group was determined to be type Am.     

Determination of MN blood group from blood stains by electrophoresis and immunoblotting

The determination of blood groups from blood stains is extremely important in medicolegal practice, but there is the possibility of an error in the determination of MN phenotypes by the absorption-elution test. We investigated a new method applying electrophoresis and immunoblotting. As a consequence of various experiments, the most appropriate pretreatment of blood stains was as follows. Blood stains were immersed in physiological saline for 0.5 to 1 h and centrifuged. The supernatant was discarded. The sediment was dissolved in sample buffer (TRIS-buffered physiological saline containing 2% sodium dodecyl sulfate) and followed by thermodegradation. It was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After transfer to a nitrocellulose membrane by Western blotting, MN phenotypes could be determined accurately from blood stains by an enzyme immunoassay (EIA) using commercially available polyclonal anti-M and anti-N sera. For blood stains more than 1 month old it was not easy to determine the MN phenotypes.     

A1 and A2 blood group substances: are there structural differences?

Blood group substances were extracted from erythrocytes of blood groups A1 and A2 and separated using HPTLC. Identification of blood group A was carried out on the chromatogramm using the PAP-technique. The specific demonstration of the serologically active glycosphingolipids allowed a comparison of the A1 and A2 blood group substance bands. The difference between the A1 and A2 Substances appears to be only a quantitative one.     

Determination of ABO(H) blood group substances from finger and toe nails

Thirty-six finger and toe nails were analyzed for ABO(H) blood group substances by the modified absorption elution method. The blood groups from nails were successfully determined in all the samples.     

Enzyme-linked immunosorbent assay for A and B water soluble blood group substances

The conditions affecting an enzyme-linked immunosorbent assay for salivary blood group substances were investigated. It was found that A, B, and O secretor saliva samples would each bind both anti-A and anti-B typing reagents. The conditions that affected the assay response were optimized for maximum sensitivity and to give the highest resolution possible between the result for an antiserum binding to homologous antigen and the response for heterologous antigen-antibody combinations. Monoclonal antibodies eliminated the heterologous binding indicating that this binding was due to a lack of specificity of the routine typing reagents. A sensitive assay using the monoclonal antibodies to distinguish between samples of A and B secretor saliva is described.     

A new 39-plex analysis method for SNPs including 15 blood group loci

A novel 39-plex typing system for single nucleotide polymorphisms (SNPs) has been developed. This multiplex approach has the advantage of being able to type 38 autosomal SNPs and one sex-discriminating base exchange site on the X and Y chromosomes rapidly and simultaneously. The SNP loci on the autosomes, which we examined, contain 15 loci distributed on blood type genes: three on RhCE, two each on Km and Gc, and one each on Duffy, AcP1, Tf, MN, GPT, EsD, PI, and Kidd genes. Thirty-seven genomic DNA fragments containing a total of 38 SNPs and one sex-discriminating site were amplified in one multiplex PCR reaction. Following the reaction, single nucleotide primer extension reaction was performed by dividing these SNP loci into five groups. The SNP type of each of the 39 loci was determined at one time by capillary electrophoresis using the newly designed multi-injection method. The combined PD (power of discrimination) of this typing system was (1-1.1) x 10(-14), and the MEC (mean exclusion chance) was 0.9990. We applied this system to forensic cases, including 16 paternity testing cases (13 non-exclusion and three exclusion cases) and one personal identification case. For the paternity testing cases, the highest Essen-M?ller's W-value was 0.9999995. The pM (matching probability) of the personal identification case was 2.22 x 10(-17). These data showed that this system was an excellent tool for use in forensic cases of paternity testing and personal identification.