1 immunohistochemistry techniques. This is one of the more reliable methods for diagnosing the disease and can be used to detect the distribution of PRRSV antigens. Halbu, et al. (1994, 1995) reported an avidin-biotin-immunoperoxidase method for detecting PRRSV antigen in the lungs of sick pigs; intranasal inoculation of 105.8TCID50ATCCVR-2386 strain at 4 heads of 3 weeks old Piglets fed colostrum were culled on the 4th or 9th day after infection, and the established artificial immunohistochemical technique was used to detect the artificial. The lung, heart, tonsil, spleen, lymph node and thymus tissues were detected as PRRSV antigen positive. Pol et al (1991) used immunoperoxidase staining to detect frozen sections of lung and spleen tissue of artificially intranasally inoculated 6-day-old SPF piglets, and the antigen was positive.
2 immunocolloidal gold technology (DIGFA). Mager et al. (1993) established an immunogold-silver staining method (1GSS) for the detection of PRRSV antigen using the specific monoclonal antibody SDOW-17, and intranasal artificial inoculation with 106.2TCID50LHVA-92-1 and 104.6TCID50LHVA-92-2PRRSV strains, respectively. Three-week-old piglets were culled on the 6th or 8th day after artificial infection. As a result, PRRSV antigen was detected in alcohol-fixed, formalin-fixed and cryopreserved lung tissues, on the 4th and 6th day after infection. On the 8th day, antigen was detected in tissues such as tonsil, lymph nodes, thymus, and kidney fixed by alcohol.
3 immunofluorescent antibody staining technique. Benfield et al. (1992) prepared a monoclonal antibody against the PRRSV nucleocapsid protein that reacted with 45 isolates from North America and Europe, indicating that this monoclonal antibody is a conserved epitope for these isolates. . An immunofluorescent antibody staining technique was established for the direct detection of antigens in pig lung tissue using fluorescein isothiocyanate (FITC) labeling. Some directly collect alveolar macrophages (PAM) from piglets for culture and diagnosis with virus-specific fluorescent antibodies.
Some of the above pathogen detection methods require cell culture, large workload, long cycle, and some need to do tissue section or even fluorescence microscope, which is not conducive to the promotion and application of the base layer detection unit, so it is rarely used in diagnosis.
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