• Korean Journal of Veterinary Service
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• v.25 no.3
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• pp.295-297
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• 2002

This experiment was done to set up the immunohistochemical detection method for infectious hematopoietic necrosis virus(IHNV) antigens in the monolayers of CHSE-214 cell cultures inoculated with IHNV. Specific identification of IHNV antigens was detected in the cytoplasms of infected cells by the use of monoclonal antibodies to glycoproteins. The specific positive signal was observed as a distinct red color. The result showed that streptavidin alkaline phosphatase immunohistochemistry specifically identified IHNV antigens in infected cultured cells.

• Korean Journal of Veterinary Service
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• v.13 no.1
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• pp.54-63
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• 1990

We have studied about the safety, immunity and protective potency in swine and experimental am mais of two inactivated vaccine produced with NYJ-1-87 strain of ADV that was isolated in Korea. Result obtained through the experiments were summarized as follows. 1. The safe potency of ADV antigens inactivated with BEI and formaline to mouse & guinea pig was on the whole good, but protective potency rates of those to challenge with ADV were 60-75％ without the differences to two antigens. 2. Safety, immunity & protective potency of ADV antigens inactivated with BEI and formaline to swine were on the whole excellent, except for a mild increase of rectal temperature in some pigs after challenge with ADV. 3. When virus excretion of the experimental groups after challenge with ADV was examed by swabbing of nasal, all pigs of control gorup excreted virus from 2 days p.c., partially to 10 days p.c.. But in vaccinated groups, only 25-50% of all pigs of each group excreted virus during experimental periods. 4. Titers of antibodies in swine & quinea pig vaccinated with inactivated ADV antigens become increased after the 1 weeth p.i. showing the highest liters on the 4-5 weeths p.i.

• Korean Journal of Veterinary Service
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• v.23 no.3
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• pp.301-306
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• 2000

In an outbreak of acute porcine diarrhea in newborn piglets, an etiological study was carried out using piglets submitted in Cheju Province Institute for Livestock Promotion(Cheju Veterinary Service for the disease diagnosis). Sixteen piglets(2-7 days old) were collected from 4 farms during outbreaks of diarrhea disease(from January to April 2000). Specimens were taken after necropsy and examined by immunohistochemistry using of monoclonal antibodies for porcine epidemic diarrhea(PED) virus, transmissible gastroenteritis(TGE) virus, and porcine rotavirus. Immunohistochemistry showed that PED virus antigens, but both TGE virus and rota virus antigens not, were localized in the some epithelial cells of the intestines of 14 animals among 16 piglets examined. PEB virus antigens were mainly detected in the cytoplasm of enterocytes. Infected cells, which were most abundant in the villous epithelial cells of the jejunum and ileum, were uncommon in the crypt, epithelial cells, the lamina propria and Peyer's patches of piglets examined. The results suggest that PED virus is one of the most prevailing agents in an outbreak of fatal diarrhea in newborn piglets on Cheju island and PED virus was need to further study to prevent this disease.

• Korean Journal of Veterinary Research
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• v.30 no.3
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• pp.297-307
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• 1990

To establish an agar-gel immunodiffusion (AGID) test for detection of antibodies to Aujeszky's disease virus(ADV) in swine, the precipitating antigens were prepared by four procedures using the Aujeszky's disease virus, NYJ-1-87 strain isolated from the affected piglets in Korea. The optimal condition for AGID test and the properties of the antigens were investigated. To determine the optimal concentration of antigens, four antigens were experimentally prepared by concentrating the viral fluids by 1/30 to 1/200. It was proved that the antigen precipitated with ammonium sulfate at concentration of 1/100 was the most efficient to detect ADV antibodies by AGID test. When the relationship between the concentration of the antigens and the size of precipitating in radial immunodiffusion test was investigated, a high correlation coefficiency at r=0.95 (y=0.23x+23.4) was estimated, In study on the effects of various buffered salt solutions and agars on the sensitivity of AGID test by using the experimental ADV antigens, it was found that 0.05M tris buffer without sodium chloride at pH 7.2 induced the most distinctive precipitating lines, and that there was no significant differences in the sensitivity between the agarose and Noble's special agar. When the efficiency of AGID test was compared with serum neutralization(SN) test, the sensitivity of AGID test was 100% in SN titer over 1 : 16, 91.7% in SN titer of 1 : 8 and 57.1% in SN titer of 1 : 4. The specificity of AGID test compared with the sera with SN titer under 1 : 2 was 98.4%. Protein analysis of the antigens by SDS-PAGE indicated that antigen I and antigen III showed a specific band of polypeptides with molecular weight of 116 K in comparison with the control antigen. Antigen IV, treated with tween-80 and ammonium sulfate, revealed specific polypeptides bands at the molecular weights 45K, 98K and 150 K.

• Korean Journal of Veterinary Service
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• v.15 no.2
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• pp.184-194
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• 1992

This study was done to identify Newcastle disease virus(NDV) antigens in paraffin sections of various organs from experimentally NDV-infected chicken using peroxidase-antiperoxidase(PAP) technique. Sections were Incubated with rabbit anti-NDV polyclonal as first antibody, followed by incubation with goat anti-rabbit IgG conjugate and peroxidase anti-peroxidase ( PAP ). Positive reactions were often detected in the epithelim of trachea and in the lymphocyte of spleen at 24 hours after virus inoculation. The viral antigen was localized mainly in the cytoplasm of infected cells. The method approved to be highly specific for the indetification of NDV and allowed a precise localization of the viral antigens in infected cells.

• Korean Journal of Veterinary Service
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• v.27 no.1
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• pp.63-73
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• 2004

The present experiment was carried out to study the pathogenesis of Newcastle disease(ND), ND virus (NDV) antigens and genes in various organs from NDV inoculated chickens were detected by immunohistochemistry and RT-PCR. Immunohistochemically, NDV antigens were detected in the spleen, thymus, cecal tonsil, proventriculus, trachea and lungs at 12 hour post-inoculation (hpi). Viral antigens were localized mainly in the cytoplasm of lymphocytes and macrophages. After 48 hpi, clinical findings of the affected chickens were open-mouth breathing, conjunctivitis, watery diarrhea and edema around the eye and neck. After 72 hpi, chickens showed muscular tremor, paralysis of the legs and wings, and coma. Histopathological results consist of multi-focal necrosis with hemorrhages in lymphoid aggregates of the intestinal tracts, necrosis of the lymphoid tissues, neuronal degeneration and necrosis, and perivascular cuffing. Using RT-PCR, virus genes were detected in the spleen and proventriculus at 48 hpi, and in the brain at 60 hpi.

• Korean Journal of Veterinary Research
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• v.29 no.4
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• pp.541-548
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• 1989

The present study was carried out to determine viral antigens and its morphogenesis in the ultrathin frozen and araldite sections of cell cultures infected with ADV by protein A-gold labeling. ADV antigens were labeled with 10nm gold probes, and electron-dense gold particles were mainly present on viral nucleocapsids and viral envelopes. Immunogold labeling in the ultracryosections showed a very low degree of interaction with tissue structures. Immunogold labeling in the ultrathin cryosections can be useful tool for the detection of ADV antigens, and the technique also may provide its great potential for immunocytochemical studies on various virus-host cell Interactions.

• Korean Journal of Veterinary Research
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• v.28 no.2
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• pp.365-369
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• 1988

The present study was done to demonstrate ADV antigens in frozen and paraffin sections from ADV-infected pigs and cell cultures by using of the IGS method. Tissue specimens from 3 young pigs infected with ADV-phylaxia strain and of 2 healthy pigs were used. Fibroblastic cells originated from pig brain and BHK cells were grown and confluent monolayers were infected with the virus. Two monoclonal antibodies and a specific hyperimmune serum to ADV were used as the source of primary antibodies for both the IGS and immunoperoxidase methods. Application of the IGS method yielded a black fine granular reaction in positive areas, and the results were superior to those obtained using the immunoperoxidase technique for all cases tested. The IGS method might be useful in the detection of various viral antigens in tissue sections.

• Korean Journal of Veterinary Research
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• v.34 no.3
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• pp.529-536
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• 1994

To establish more specific and simple diagnostic methods for detection of the antibodies and antigens of Aujeszky's disease virus(ADV), we designed indirect dot-immunoassay(IDI) and double sandwich dotimmunoassay(DSDI) using the solid phases of nitrocellolose paper and polystyrene plate. The diagnostic efficacy of these methods was investigated. As the sensitivity of IDI was tested by various virus concentration, the specimens with the virus titer above $10^{4.0}TCID_{50}/0.2ml$ showed positive reaction, but that below $10^{1.0}TCID_{50}/ml$ revealed negative. Tonsil emulsion at the virus titer of $10^{4.5}TCID_{50}/0.2ml$ showed the highest sensitivity as diluted by 1/100. In detection of ADV antigens from the various tissues of the rats and pigs infected with ADV, IDI using monoclonal antibody showed the higher specificity as compared with IDI using polyclonal antibody and virus isolation method. The efficacy of the DSDI for detection of ADV antibody was compared with other tests. The sensitivity of DSDI was higher than virus neutralization(VN) and agar gel immunodiffusion test(AGID). Meanwhile, specificity of DSDI was lower than AGID, but similar to IDEA. In comparison with VN test, DSDI showed 96.9% agreement to VN test that is the highest of three tests. In general, application of polyclonal antibody in both tests caused the higher sensitivity but the lower specificty.

• Korean Journal of Veterinary Pathology
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• v.3 no.2
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• pp.73-76
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• 1999

The tissue distribution and cellular localization of viral antigen in the brain of aborted fetus with bovine viral diarrhea virus(BVDV) infection was studied; BVDV antigens was detected in spleen, kidney, lung, eyelid as well as brain. In the brain, the virus was recognized in neurons and non-neuronal cells in the cerebellum and cerebrum. Many cells in the superficial layer and occasional Purkinje cells had BVDV antigens. As well, BVDV was also found in the perivascular cells, vascular endothelial cells and smooth muscle cells in the vessels and neuroglial cells in the white matter. This finding suggests that BVD virus favors infect progenitor cells in the brain, notably in the superficial layer of cerebellum, and damage normal development of cerebellum, which leads to cerebellar hypoplasia.