| In this experiment, BALB/c mice were immunized with purified recombinant PRV VP6 protein, hybridoma supernatants were screened by indirect ELISA, using the hybridoma technique and the limiting dilution, one hybridoma cell line named C5D10 w was obtained after four-time subclone , which could secrete monoclonal antibody(McAb) specific against PRV. The isotype of the McAb was IgG1, the average number of chromosomes was 93,the ELISA titers of culture supernatants and ascites were 1 : 800 and 1 : 1 ×106, respectively. The McAb didn,t react with transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea coronavirus (PEDV) and pseudorabies virus (PrV). The results show that the C5D10 McAb has high specificity to PRV. The Marc-145 cell infected with PRV was carried out comparative studies which is detection of pathogen with indirect immunofluorescence using monoclonal antibodies against PRV VP6 protein and polyclonal antiserum against PRV. The result indicates that Marc-145 cells infected with PRV shows strongly green fluorescent in cytoplasm and cell membrance with indirect immunofluorescence by McAbs, the cells uninfected PRV are stained red by Evance blue and there are no green fluorescent cells. The cells infected with PRV also shows strongly green fluorescence; but the cell uninfected with PRV exists certain non-specific fluorescent response. The specificity result of C5D10 McAbs which react to others diarrhea virus and the IFA test suggest C5D10 McAb is highly specific to PRV. All the work founded material base for PV diagnosis accurately and rapidly.
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