Detection of infectious bovine rhinotracheitis virus by immunofluorescence and polymerase chain reaction
By: Anupama K.
Contributor(s): Koshy John(Guide).
Material type:
BookPublisher: Mannuthy Department of Veterinary Microbiology, College of Veterinary and Animal Sciences 2008Description: 72.DDC classification: 636.089 6 Online resources: Click here to access online Dissertation note: MVSc Abstract: A study was undertaken to detect the presence of IBR virus in clinical samples by PCR and immunofluorescence and to compare the efficacy of these tests. Attempts were also made to isolate the virus from clinical samples in MDBK cell line.
A total of 60 samples from suspected animals and 5 samples from healthy animals were collected from various sources like, University Livestock Farms, Veterinary Hospital, Mannuthy and some private dairy farms in Thrissur district. The samples included nasal and vaginal swabs, aborted materials and semen. All these samples were screened for the presence of IBR virus by PCR and immunofluorescence. The TK gene based primers were used for PCR and the positive control generated an amplicon of size 298bp. No sample was found positive by PCR. Immunofluorescence was performed on smears prepared from clinical samples using FITC conjugated polyclonal antibovine IgG. This test detected viral antigen in one of the samples, indicated by focal, bright yellowish green fluorescence associated with the cytoplasm of epithelial cells. The inability of PCR for detection of viral DNA in sample tested positive by immunofluorescence may be due to the presence of non-specific inhibitors in clinical sample. These results suggest that even though PCR is highly sensitive compared to other diagnostic tests, false negative results cannot be excluded. Immunofluorescence on smears prepared from clinical samples can be considered as a rapid diagnostic method for IBR. Attempts to isolate the virus from clinical samples in MDBK cell line were found unsuccessful.
| Item type | Current location | Call number | Status | Date due | Barcode |
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Theses
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KAU Central Library, Thrissur Theses | 636.089 6 ANU/DE (Browse shelf) | Available | 172854 |
MVSc
A study was undertaken to detect the presence of IBR virus in clinical samples by PCR and immunofluorescence and to compare the efficacy of these tests. Attempts were also made to isolate the virus from clinical samples in MDBK cell line.
A total of 60 samples from suspected animals and 5 samples from healthy animals were collected from various sources like, University Livestock Farms, Veterinary Hospital, Mannuthy and some private dairy farms in Thrissur district. The samples included nasal and vaginal swabs, aborted materials and semen. All these samples were screened for the presence of IBR virus by PCR and immunofluorescence. The TK gene based primers were used for PCR and the positive control generated an amplicon of size 298bp. No sample was found positive by PCR. Immunofluorescence was performed on smears prepared from clinical samples using FITC conjugated polyclonal antibovine IgG. This test detected viral antigen in one of the samples, indicated by focal, bright yellowish green fluorescence associated with the cytoplasm of epithelial cells. The inability of PCR for detection of viral DNA in sample tested positive by immunofluorescence may be due to the presence of non-specific inhibitors in clinical sample. These results suggest that even though PCR is highly sensitive compared to other diagnostic tests, false negative results cannot be excluded. Immunofluorescence on smears prepared from clinical samples can be considered as a rapid diagnostic method for IBR. Attempts to isolate the virus from clinical samples in MDBK cell line were found unsuccessful.
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