浅在性白癬1466例について分布, さらに病型や原因菌種別の血液型分布を調査したが, 日本人の血液型分布との間に有意の差はみられなかった。浅在性自癬ととは関連がないと考えられた。

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ABO type test was given to 469 lepers, and S type to 406 without finding any specific relation between abnormality types and constitution. Survey on 48 groups of lepers among kinship revealed no specific relationship to the disease.

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  Universal immuno-peroxidase polymer (UIP) technique on the ABO blood grouping was applied to samples of 4 μm thick cross sections obtained from human scalp and pubic hairs by using a microtome.
  The UIP technique consists of amino acid polymers which are conjugated to multiple molecules of peroxidase and anti-mouse Immunoglobulin that is reduced to its Fab' fragment. Advantages of the UIP technique are: a) more intense staining, b) skipping two steps of staining and c) lower background staining. This technique was carried out using monoclonal antibodies, mouse anti-A (diluted 1:10), anti-B (diluted 1:10) and anti-H as the primary antibodies. With this technique, specific staining was shown as dark brown precipitates within the medulla of hair samples, and depending on the staining, respective blood groups were able to be determined.
  From this investigation, it seemed that the present technique was of practical use for the ABO blood grouping of a sample from human scalp and pubes hairs.

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  For ABO blood typing of human salivary stains, a new mixed agglutination reaction method was devised, in which, OHP film was used as antigen-antibody reaction plate. By this method a series of examination procedures such as extraction of blood group substances, heating-fixation of the substance and incubation with monoclonal antibodies could be performed successively on the OHP film. For the secretor's salivary stain samples, specific agglutination reaction which presents an agglutination circle was observed. ABO blood typing from secretor's salivary stains was correctly performed by this method.

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There are three common alleles (A, B, and O) at the human ABO blood group locus which codes glycosyltransferase. The polymorphisms of the ABO blood group are also observed in wide variety of primates. The difference of the glycosyltransferase activity between human A and B enzymes is due to the two amino acid differences. The same amino acid differences are observed for A and B alleles in non-human primates. We determined 19 sequences of chimpanzee, 8 sequences of bonobo, and 2 sequences of Japanese macaque ABO blood group gene for exon 3 and intron 6 (ca 1.7kb). We also determined 3 sequences of Japanese macaque ABO for exon 7 (ca. 0.5kb). We compared those data with published sequences of other hominoids and Old World monkeys. It was suggested that the type changes between A and B occurred independently in the both lineages of the hominoids and Old world monkeys. The alleles A and B appeared to be polymorphic in the ancestral species of macaques, while the different B type allele evolved independently in baboon lineage.

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  A highly sensitive method for forensic ABO blood grouping using monoclonal antibodies was developed in combination with an absorption-elution test and an enzyme-linked immunosorbent assay (ELISA). In this method, monoclonal antibodies eluted from bloodstains with the absorption-elution, were determined by ELISA, for detection of ABO blood group substances in bloodstains. This method enabled us to determine blood group substances in bloodstains with high sensitivity about 10 times greater than a usual absorption-elution test and also to handle many samples (32 samples per a ELISA plate) at the same time. Furthermore, changing the heat elution of antibodies to the acid elution permitted automated analysis for ABO blood grouping using BIOMEK 1000 apparatus (Beckman Instrument, California, U. S. A.).

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  The effect of pH on color development was examined in detail to detect higher sensitivity of ABO blood groups substances in saliva and semen by means of an indirect enzyme-linked immunosorbent assay (ELISA) using secondly antibody conjugated with horseradish peroxidase. The use of McIlvaine buffer was the most effective for color development. Deep color was observed at pH 5.0, and intensity was about 32 times stronger than that obtained with the Tris-HCl buffer (pH 7.6) that was widely used in the Japanese Criminal Investigation Laboratories. Owing to the increased sensitivity, B-type and O-type blood of non-secretors semen, which could not be detected by ELISA, were successfully determined under the present conditions. Through this study, we found the interesting fact that commercially available monoclonal antibodies can be divided into two groups: One group reacts with secretor sample alone and the other reacts with both secretor and non-secretor samples. Using these two classified antibodies, we were able to discriminate between secretor and non-secretor and to speculate the blood type of one sample from two mixed samples.

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  Application of acid elution to absorption-elution test for ABO blood-typing using commercial monoclonal antibodies was described. Collodion plates to fix blood or body fluid samples were introduced for the convenience of treatment of a lot of test samples. For evaluating the optimum elution condition, the amounts of the antibodies eluted were semi-quantitatively determined using enzyme-linked immunosorbent assay (ELISA). The optimum pH for elution was between 3.0 and 3.5 and a remarkable deactivation of antibodies under strong acid condition was observed. Therefore, the accurate pH control was required for effective elution. Elution time was decided at 10 min because the active antibodies were eluted within a few minutes and were maintained during subsequent 20 min. Under these optimum conditions, the absorption-elution test with indicator cells was achieved using some antibodies suitable for acid elution. We also obtained a good result at the preliminary test for an automatic blood-typing system using collodion plate and the acid elution method combined with ELISA. The technique described in this report was considered to be helpful to establish a full automatic blood-typing system for forensic specimen.

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