Enzyme Combined Sorption Assay

Enzyme-linked immunosorbent assay (hereinafter referred to as ELISA) is the most widely used technique in enzyme immunoassay. It is used to detect the antigen (or antibody) to be tested which is coated in the well of the solid phase plate. That is, the antibody is labeled with an enzyme, and the known antigen or antibody is adsorbed on the surface of the solid phase carrier, and the antigen-antibody reaction is carried out on the surface of the solid phase carrier, and the free component in the liquid phase is washed away by washing, and finally acted on by the enzyme. The color is developed after the substrate to judge the result. Basic Information Specialist classification: growth and development check classification: biochemical examination Applicable gender: whether men and women apply fasting: fasting Tips: Obey the doctor's instructions when checking. Normal value The ratio of the serum to be tested to the known negative serum (P/N) was <2.1, and the OD value of the serum to be tested was <0.4, which was judged to be negative. Clinical significance Abnormal results: the ratio of the serum to be tested to the known negative serum (P/N) ≥ 2.1, and the OD value of the serum to be tested is ≥ 0.4, it is considered positive. Need to check the crowd: detect antigen and antibody. Precautions Contraindications before inspection: Prepare various packaging solutions and set the concentration. Taboo when checking: 1. Strictly control factors affecting the efficiency of labeling such as temperature, time, pH, enzyme and antibody amount. 2. When installing the column, make the column uniform, the cylinder surface is flat, no bubbles or cracks. Inspection process Commonly used ELISA methods include a double antibody sandwich method and an indirect method, the former for detecting macromolecular antigens and the latter for measuring specific antibodies. Indirect ELISA This method is mainly used to detect antibodies. The procedure for indirect ELISA is as follows. (1) Materials 1 coating liquid, washing liquid, heat preservation liquid, substrate liquid, and stopping liquid. 2DVH coated antigen, enzyme-labeled anti-antibody, negative and positive DVH reference serum. 3 ELISA detector, sampler, polystyrene microplate. (2) Method steps 1 plus antigen coating → 4 ° C overnight, washed three times, throw dry. 2 plus serum to be tested → 37 ° C for 2 hours, washed three times, throw dry. 3 Add the enzyme-labeled antibody → 37 ° C for 2 hours, wash three times, throw dry. 4 Add substrate solution → 37 ° C for 30 minutes, add stop solution. 5 The OD value was measured by an ELISA detector, and the P/N ratio was calculated. 2. Double antibody sandwich ELISA This method is mainly used to detect macromolecular antigens. 1 Add antibody coating → 4 ° C overnight, wash three times, throw dry. 2 Add the antigen to be tested → 37 ° C for 30 minutes, wash three times, throw dry. 3 Add the enzyme-labeled antibody → 37 ° C for 30 minutes, wash three times, throw dry. 4 Add substrate solution → 37 ° C for 15 minutes, add stop solution. 5 The OD value was measured by an ELISA tester. Not suitable for the crowd Not suitable for the crowd: no. Adverse reactions and risks No related complications or hazards.