Central Veterinary Diagnostic Laboratory

Tando Jām, Pakistan

Central Veterinary Diagnostic Laboratory

Tando Jām, Pakistan
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Sharawi S.S.A.,Banha University | Sharawi S.S.A.,Central Veterinary Diagnostic Laboratory | Al-Hofufy A.N.,Central Veterinary Diagnostic Laboratory | Al-Blowi M.H.,Central Veterinary Diagnostic Laboratory
International Journal of Virology | Year: 2011

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. © 2011 Academic Journals Inc.

Lu H.,Pennsylvania State University | Ismail M.M.,King Faisal University | Ismail M.M.,Kafr El Sheikh University | Khan O.A.,Texas A&M University | And 3 more authors.
Avian Diseases | Year: 2010

The first outbreak of H5N1 highly pathogenic avian influenza (HPAI) in the Kingdom of Saudi Arabia (KSA) occurred in two "backyard" flocks of Houbara bustards and falcons in February 2007. Subsequent outbreaks were seen through the end of 2007 in "backyard" birds including native chickens, ostriches, turkeys, ducks, and peacocks. From November 2007 through January 2008, H5N1 HPAI outbreaks occurred in 19 commercial poultry premises, including two broiler breeder farms, one layer breeder farm, one ostrich farm, and 15 commercial layer farms, with approximately 4.75 million birds affected. Laboratory diagnosis of all H5N1-positive cases was conducted at the Central Veterinary Diagnostic Laboratory (CVDL) in Riyadh, Saudi Arabia. A combination of diagnostic tests was used to confirm the laboratory diagnosis. A rapid antigen-capture test and real-time reverse transcriptase-PCR (rtRT-PCR) assay on clinical and field specimens were conducted initially. Meanwhile, virus isolation in specific-pathogen-free embryonating chicken eggs was performed and was followed by hemagglutinin (HA) and hemagglutination inhibition tests, then rapid antigen-capture and rtRT-PCR tests on HA-positive allantoic fluid samples. In most HPAI cases, a complete laboratory diagnosis was made within 24-48 hr at the CVDL. Saudi Arabian government officials made immediate decisions to depopulate all H5N1-affected and nonaffected flocks within a 5-km radius area and applied quarantine zones to prevent the virus from spreading to other areas. Other control measures, such as closure of live bird markets and intensive surveillance tests on all poultry species within quarantine zones, were in place during the outbreaks. As a result, the HPAI outbreaks were quickly controlled, and no positive cases were detected after January 29, 2008. The KSA was declared free of HPAI on April 30, 2008, by the World Animal Health Organization. © 2010 American Association of Avian Pathologists.

Nizamani A.R.,Sindh Agriculture University | Nizamani Z.A.,Sindh Agriculture University | Umrani A.P.,Central Veterinary Diagnostic Laboratory | Dewani P.,Central Veterinary Diagnostic Laboratory | And 3 more authors.
Journal of Animal and Plant Sciences | Year: 2015

Peste des petits ruminants (PPR) is an acute viral disease of small ruminants which is clinically characterized by occulonasal discharges, oral erosive lesions, diarrhea, pneumonia and leucopenia.It leads to high morbidity and moderate mortality rates.The present study was aimed at determining the seroprevalence of PPR in small ruminants of Sindh province by competitive-ELISA. A total of 7096bloodsamples were collected from sheep (1309) and goats (5787) fromeleven districts of the Sindh. The overall seroprevalence was found to be35.23 percent in the small ruminant population.Prevalence was found higher in sheep (37.2%) than in goats (34.78%). Seroprevalence varied in various age groups and was found to be 33.41%, 33.34% and 39.15% in three age groups, i.e.<1 year, 1 to 2 years and > 2 years, respectively.Prevalence rates were higher in females (35.94%) than in males (31.23%). Prevalence of PPRvaried in various agro-ecological zones of the province. It was found highest in Dadu district (69.3%) and lowest in Badin district (15.7%). Southern coastal districts of Badin (15.7%) and Thatta (30.9%) showed lower and eastern districts of Tando Allah Yar (60.3%) along with Tharparkar (50.3%) a higher seroprevalence of PPR. PPR is endemic in Sindh, therefore there is an urgent needfor preventive vaccination disease control. © 2015, Pakistan Agricultural Scientists Forum. All rights reserved.

Habib M.,Nuclear Institute for Agriculture and Biology | Shah M.S.,Nuclear Institute for Agriculture and Biology | Muzammil H.M.,Nuclear Institute for Agriculture and Biology | Manzoor S.,Model Civil Veterinary Hospital | And 4 more authors.
Pakistan Journal of Life and Social Sciences | Year: 2014

Sampling was done from various outbreaks of foot-and-mouth disease (FMD) that occurred during the year 2013 in district Faisalabad, Punjab, Pakistan. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used for typing of foot-and-mouth disease virus (FMDV). A total of 110 clinical samples were received from different administrative regions of Faisalabad; 86 were found FMDV genome positive when tested with 1F and 1R consensus primers. Out of these genome positive samples, 73 were successfully typed into serotypes A (19 samples), O (30 samples) and Asia1 (24 samples). Complementary DNAs (cDNAs) were stored at - 80°C for further sequencing studies. Serotype A FMDV was detected from samples received from outbreaks that occurred in Sumandri, Jaranwala and Tandlianwala tehsils, serotype O from Tandlianwala and Sadhar, and serotype Asia 1 outbreaks were reported from Chak Jhumra and Sadhar.

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