<br> <br> immune staining test to detect diseased tissues viral antigen directly labeled antibody. The cortical cells of the kidney infected with swine fever are stained dark brown. Application contains o. 01% hydrogen peroxide or o. 01% sodium azide solution inhibited endogenous enzymes and achieved good results.
The diseased pig tonsil, spleen, kidney, lymph nodes and other diseases were embossed or frozen. After the specimen was naturally dried, 10rnin was fixed in a mixture of 2% glutaraldehyde and formaldehyde. After drying, the specimen was immersed in Tris-HCI buffer containing 0.01% hydrogen peroxide or 0.01% sodium azide solution and allowed to act at room temperature for 30 min. Rinse 5 times with pHL 40.02 mol/L phosphate buffer for 3 min each time, then air dry. Place the specimen in a wet box, add 1:10 enzyme-labeled antibody, 3 min at 37~C: rinse with pH 7.40.02mol/L phosphate buffer for 3~4 times, each time for 2~3min, then use no Water ethanol, xylene dehydration, sealing inspection. Outcome judgment: Microscopic examination, such as cells are dark brown positive; yellow or five colors are negative. The cytoplasm of pig tissue inoculated with attenuated pigs was slightly brown, which was significantly different from the infection of virulent strains.
IFA <br> <br> be divided into direct and indirect immunofluorescence immunofluorescence test.
Tissue direct immunofluorescence assay was used to collect typical suspicious tissues or organs, embedded in paraffin, sliced ​​or frozen, and sliced ​​with a cryostat. The cells were fixed with acetone at room temperature for 10 min and the tissue pieces were air-dried. Then, the tissue pieces were washed with PBS. The antibody was incubated (indirect fluorescence was performed in two steps, direct fluorescence was one step), and the tissue pieces were washed again with PBS, and the results of fluorescent staining were observed under a fluorescence microscope after air drying. The method has the advantages that it does not need to collect a large amount of diseased parenchyma, and the operation is simple. Disadvantages: low specificity and sensitivity, subjective.
Indirect immunofluorescence test: 1:4 anti-porcine sputum positive serum was added to the diseased pig tissue, and after 30 mln, the goat anti-porcine IgG fluorescent antibody was added and washed for 30 minutes, rinsed, dried and microscopically examined. Advantages: easy to operate and quick diagnosis. Disadvantages: costly, poorly specific, subjective, requires an experienced inspector to distinguish between non-specific staining.
The diseased pig tonsil, spleen, kidney, lymph nodes and other diseases were embossed or frozen. After the specimen was naturally dried, 10rnin was fixed in a mixture of 2% glutaraldehyde and formaldehyde. After drying, the specimen was immersed in Tris-HCI buffer containing 0.01% hydrogen peroxide or 0.01% sodium azide solution and allowed to act at room temperature for 30 min. Rinse 5 times with pHL 40.02 mol/L phosphate buffer for 3 min each time, then air dry. Place the specimen in a wet box, add 1:10 enzyme-labeled antibody, 3 min at 37~C: rinse with pH 7.40.02mol/L phosphate buffer for 3~4 times, each time for 2~3min, then use no Water ethanol, xylene dehydration, sealing inspection. Outcome judgment: Microscopic examination, such as cells are dark brown positive; yellow or five colors are negative. The cytoplasm of pig tissue inoculated with attenuated pigs was slightly brown, which was significantly different from the infection of virulent strains.
IFA <br> <br> be divided into direct and indirect immunofluorescence immunofluorescence test.
Tissue direct immunofluorescence assay was used to collect typical suspicious tissues or organs, embedded in paraffin, sliced ​​or frozen, and sliced ​​with a cryostat. The cells were fixed with acetone at room temperature for 10 min and the tissue pieces were air-dried. Then, the tissue pieces were washed with PBS. The antibody was incubated (indirect fluorescence was performed in two steps, direct fluorescence was one step), and the tissue pieces were washed again with PBS, and the results of fluorescent staining were observed under a fluorescence microscope after air drying. The method has the advantages that it does not need to collect a large amount of diseased parenchyma, and the operation is simple. Disadvantages: low specificity and sensitivity, subjective.
Indirect immunofluorescence test: 1:4 anti-porcine sputum positive serum was added to the diseased pig tissue, and after 30 mln, the goat anti-porcine IgG fluorescent antibody was added and washed for 30 minutes, rinsed, dried and microscopically examined. Advantages: easy to operate and quick diagnosis. Disadvantages: costly, poorly specific, subjective, requires an experienced inspector to distinguish between non-specific staining.
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