Veterinary Archives, Vol. 82 No. 5, 2012.
Original scientific paper
Fluorescent in-situ hybridization technique for the detection and localization of classical swine fever virus in infected tissues.
Kumaresan Nagarajan
orcid.org/0000-0001-6420-0309
; Department of Veterinary Pathology, Madras Veterinary College, TANUVAS, Vepery, Chennai, Tamil Nadu, India
Guttula Saikumar
; National CSF Referral Laboratory, Division of pathology, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, India
Abstract
Classical swine fever (CSF) may be diagnosed by various conventional techniques but nucleic acid based methods have gained popularity because of their high sensitivity and specificity. For detection of viral RNA in infected tissues, a fluorescent in-situ hybridization (FISH) based method was developed using a 308 bp biotinylated DNA probe targeting E2/NS2 gene of CSFV. In-situ RNA/DNA hybrids were detected with fluorescently labeled Streptavidin conjugate and viewed under a fluorescent microscope. Viral nucleic acids could be demonstrated in the lymphoid tissues such as the spleen and lymph nodes collected from CSFV infected animals. Infected macrophages and reticuloendothelial cells revealed bright intracytoplasmic fluorescence. In conclusion, the fluorescent in situ hybridization technique used in the study provided an effective means of detection and localization of CSFV nucleic acids.
Keywords
classical swine fever virus; fluorescent in-situ hybridization; virus localization
Hrčak ID:
86323
URI
Publication date:
11.9.2012.
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