快速斑点-ELISA法测定体液(斑)血型的建立

THE REPID DETERMINATION OF ABO GROUP FROM BODY FLUIDS (OR STAINS) BY DOT-ELISA USING ENZYME-LABELLED MONOCLONAL ANTIBODY

本实验应用ABH酶标单克隆抗体,通过斑点ELISA,快速进行体液和体液斑的血型测定。在实验中,将待测抗原滴在硝酸纤维膜上,然后将膜分别放入抗A、抗B和抗H酶标单克隆抗体中,再加二氨基联苯胺底物。抗原抗体反应阳性部位将出现一个清晰的棕褐色斑点。我们共测定521份检材,结果均正确。本方法准确、简便、灵敏、快捷,结果易观察,不需特殊仪器设备,30分钟内即可获得结果。其灵敏度比常规的中和试验、吸收试验高很多。10^(-4)～10^(-5)稀释的唾液、所谓非分泌型人唾液用此方法仍能测定血型。

Using ABH enzyme-labelled monoclonal antibody, we could rapidly detect ABO group from body fluids and body fluid stains by dot-ELISA. In this test, the antigen was immobilized on nitrocellulose paper and the entire piece of paper was coated with an appropriate dilution of enzyme-labelled McAb directly against the antigen of interest and finally 3, 3'- diaminobenzidine (DAB) substrate solution was added. The site of positive is clearly visible as a brown spot. We detected 521 samples and got a satisfactory result. We also detected 99 practical case samples by this method. The result is the same as using other methods by other people. This method is accurate, simple, direct, rapid, sensitive, easy to observe, requisite with no special equipment and complete in 30 minutes. The test was proved to be clearly more sensitive for the detection of ABO blood group in secretor-saliva than the conventional hemagglutination inhibition test, 10-4-10-5 dilution of saliva and ABO group of non-secretor-saliva and urine could also be easily detected by this method.