Studies on the determination of blood groups from small amounts of bloodstains by use of enzyme-linked immunosorbent assay(ELISA).

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Enzyme-linked immunosorbent assay(ELISA) or enzyme-linked fluoroimmunosorbent assay(ELFA) with an avidin-biotin complex(ABC) was applied to the determination of ABH or Lewis blood group from salivas, Bloodstains and nails, and the results were as follows.1. ABH group of small amounts of non-secretion salivas was typed precisely by the present ELISA, although that of those was not determined by a conventional absorption inhibition test.2. The Lewis typing of bloodstains could be determined by the ELISA using monoclonal anti-Lea and anti-leb antibodies. About 1 mg of the stains was enough to type each Lewis substance reliably.3. The ELFA procedure was established. About 0.1 mg of the bloodstains was enough to type each Lewis antigen by this method.4. The Lewis typing of Salivastains was done by the ELISA. Although Lea substance in Le(a-b+)type saliva was secreted in various ratios, the level of Leb substance was superior to that of Lea antigen. Likewise, the same results in Le(a+b-)type salivastains were obtained. The level of both Lea Leb substances in Le(a-b-)type salivastains was always low compared to that of the other two types. The Lewis typing, therefore, was determined easily by measuring 2 to 3 points of serial two-fold saliva dilutions.5. It was evident taht nails contain the Lewis substances. However, the level of the Lewis substances did not reflex that of the corresponding erythrocytes.6. MN blood typing in bloodstains was also tested by the ELISA, but satisfactory results were not obtained. This may be attributed to the antibodies used.7. A software was established to automate the determination of blood grouping; it took only 3 to 4 min to read the data precisely by using this software.