A series of examinations is presented for human origin identification and ABO blood grouping of doubtful minute human blood stains. A blood-stained thread (0.5 cm in length) was first tested to identify human origin by microprecipitation method and then the ABO blood type was determined by both a modified absorption-elution test and a modified mixed agglutination. In the continuous tests, the maximum limits of positive reactions of the microprecipitation method, the modified absorption-elution test, and the modified mixed agglutination were 1:640, 1:160, and 1:2,560 diluted blood, respectively. A and B agglutinogens were more sensitively determined than H agglutinogen. Hemagglutinogens of blood stains on cotton threads were more easily detected than those of polyester ones.

The presence of M and N antigens in the blood components other than the red cells and various tissues has not been settled yet . The elution method in combination with antiglobulin test reported by the present authors has been used for grouping the stains of the leucocytes , platelets, fibrin, and thymic lympho cytes. All of the test-stains showed obviously positive antigen activities which coincided with blood group of the sources. The experiments of the test-stains treated with trypsin revealed that the treated stains showed a similar agglutin ability to that of the treated red cells. From the results, it is concluded that the blood components have M or N antigen activity which is a little lower than the red cells.MMIN blood group; leucocytes; platelet; fibrin; lymphocytes It has been well admitted that M and N group antigens are on the blood red cells. As regards the blood components other than red cells and various tissues, there are two antagonistic opinions: Boorman and Dodd (1943), Kosyakov and Kuznetsova (1962) and Majsky (1960) reported that M and N group antigens were contained in various human tissues, whereas Clausen (1933) and Krah (1952) pointed out that the antigens were demonstrated only in the blood red cells. Recently, serological techniques to detect antigens or antibodies have been remarkably ameliorated. For example, the mixed agglutination or the groupspecific double combination method (Coombs and Dodd 1961, Akaishi 1965), and the absorption-elution method (Kind 1960;Yada 1962) are able to demonstrate easily the blood group of minute saliva stains belonging to non-secretor, which cannot be detected with the agglutination inhibition test. The present authors reported previously ) an improved method for MN blood grouping, which is a modified elution method in combination with anti-globulin test, and MN blood groups of very minute stains, even 2-to 3-year-old, were detected by this method. The present paper deals with M and N antigens of the leucocytes, platelets, fibrin and thymic lymphocytes. MATERIALS AND METHODSTest-stains of the leucocytes, platelets, fibrin, and thymic lymphocytes. Each of M, N and MN group blood containing standard acid-citrate-dextrose was centrifuged at 1,300

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Bloodstained threads (1 cm in length) were tested to identify human origin by a direct ELISA-ABC method using biotinylated antibody against human HbA0. By this method human bloodstains were clearly distinguishable from bloodstains of other species including Japanese monkey. The minimum detection limit of bloodstains prepared from undiluted human whole blood was 1:5, 120 (28 ng Hb) and that of bloodstains from diluted human whole blood was 1:640-1:1, 280. Human Hb was more easily detectable in bloodstains prepared from diluted human blood after extraction with 5% ammonia than after extraction with phosphate-buffered saline.

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