[Simple determination methods for blood grouping from biological stains. 1. Immunologic gold colloid aspiration test and dot ELISA test by membrane aspiration method for determining ABO blood groups from saliva specimens].

Two immunologic tests using nitrocellulose membrane and an aspiration device were applied to determine the ABO blood groups and secretor status from saliva specimens. First, immunologic gold colloid membrane aspiration test (IMAT) was performed with orange-red colloidal gold particles (GP) sensitized with anti-A or anti-B antibody or anti-H lectin (A chicken antiserum to O secretor saliva was also used as an anti-H reagent in some experiments). The boiled saliva specimens were serially diluted and dotted to the nitrocellulose membrane, which was equipped to a disposable aspiration kit, and the antibody- or lectin-coated GP was aspirated through the membrane. The stained saliva changed red by the immunologic binding of the GP. Test results could be obtained within some minutes. Detection limits of IMAT shown by saliva dilution were 1:100 to 1:10,000 for secretor salivas, whereas those for non-secretor salivas were 1:10 or lower. Second, a revised dot ELISA was performed by using the membrane aspiration device. The requisite time for this test (Dot ELISA-A) was 40 to 45 minutes, whereas that for conventional dot ELISA was 3 to 4 hours. The detection limits of the dot ELISA-A shown by the saliva dilutions were 1:1,000 to 1:100,000 for secretor salivas, whereas those for non-secretor salivas were 1:1,000 or below.