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International Journal of Virology
  Year: 2011 | Volume: 7 | Issue: 4 | Page No.: 147-157
DOI: 10.3923/ijv.2011.147.157
Innovation of Indoor Real-time Polymerase Chain Reaction for Diagnosis of Camel Pox Virus in Clinical Field Samples using Primer Site Belongs to Capripoxvirus
S.S.A. Sharawi, A.N. Al-Hofufy and M.H. Al-Blowi

Abstract:
Real-time amplification techniques are currently used to determine the viral Nucleic Acid (NA) in clinical samples for diagnostic purposes. In disease management contexts, until now, a little have been described for the molecular detection of Camel Pox Virus (CPV). This study reports the development of a Real-Time Polymerase Chain Reaction (RT-PCR) for detection of CPV using SYBR green I chemistry. A total of 15 specimens from camels suspected of being infected with CPV were collected from Riyadh Province during 2009 and submitted for virological investigation at the Central Veterinary Diagnostic Lab. (CVDL), Riyadh, Ministry of Agriculture; KSA. In solution; detection and identification of CPV was achieved in 10 samples by conventional Polymerase Chain Reaction (PCR). During further studies performed, it was shown that CPV was isolated in Chorio-allantoic Membranes (CAMs) and in Vero cell as well as demonstration of pock lesions and Cytopathic Effect (CPE) due to CPV were observed while CPV virus antigen was detected and identified by Indirect Immune-fluorescent Assay (IFAT) in Vero cells. A trial for development of a simple and rapid qualitative Real-time Polymerase Chain Reaction (RT-PCR) was applied using primer site belongs to Capripoxvirus to detect CPV load in prepared tissue samples comparing to inoculated CAMs and Vero cells [reveal pocks or CPE] where, NA of CPV in samples were (13/15) while; CAMs and Vero cells were (7/15) and (9/15) positive, respectively. The obtained SYBR Green dye-based Real-time PCR results; comparing to ordinary PCR or CPV isolation in eggs or cell cultures; proved that this development RT-PCR assay was rapid, accurate and effective for the direct and qualitative detection of CPV (viral DNA) in both necropsy specimens and inoculated egg or tissue culture samples. To the authors knowledge, this is the first report to describe a primer set of Capripoxvirus gene-based Real-time PCR for specific diagnosis of CPV infection in clinical samples.
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How to cite this article:

S.S.A. Sharawi, A.N. Al-Hofufy and M.H. Al-Blowi, 2011. Innovation of Indoor Real-time Polymerase Chain Reaction for Diagnosis of Camel Pox Virus in Clinical Field Samples using Primer Site Belongs to Capripoxvirus. International Journal of Virology, 7: 147-157.

DOI: 10.3923/ijv.2011.147.157

URL: https://scialert.net/abstract/?doi=ijv.2011.147.157

 
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