SN/T 1247.2-2003Protocol of immunoperoxidase monolayer assay for porcine reproductive and respiratory syndrome (English PDF)

3.1 AEC

Aminoethyl carbazole.

3.2 HPRO

Horseradish peroxidase.

3.3 IPMA

Immunoperoxidase monolayer assay.

3.4 PRRS

Porcine reproductive and respiratory syndrome.

4.1 Principle

The immunoperoxidase monolayer test is to adsorb an antigen on a solid phase carrier, and add a sample to be tested to form an antigen-antibody complex.

The enzyme-labeled anti-antibody is colored with the participation of the substrate, and the result is determined by visual inspection with a naked eye or a spectrophotometer after the substrate is developed.

The principle of this standard is to culture cells inoculated with porcine reproductive and respiratory syndrome virus on 96-well cell culture plates.

Into a certain proportion of diluted serum to be tested, after the end of the reaction, add anti-porcine immunoglobulin antibody labeled with HPRO, after the reaction is completed, the substrate was added and the reaction results were examined.

4.2 Sampling

Collect the blood of the pig to be tested, separate the serum, the serum must be fresh, transparent, non-hemolyzed, non-polluting, and sealed in a sterile vial at 4 ° C or-20 ° C ~ -30 ° C refrigerator storage or cold storage conditions immediately sent for inspection. The serum to be tested was numbered uniformly before the test.

4.3 Equipment and materials

4.3.1 Instrumentation

---Micropipette;

---Heat box;

---Ordinary incubator and carbon dioxide incubator;

---Ordinary refrigerator and low temperature refrigerator;

---Inverted microscope;