Application of some molecular assays in the quality control of PPR Viral Vaccine / Manar Fathy Hassan Seioudy ; Supervised Mohammed Abdelhameed Shalaby , Ahmed A. Elsanousi , Magda M. Sayed

Thesis (Ph.D.) - Cairo University - Faculty of Veterinary Medicine - Department of Virology

Peste des petits ruminants (PPR) is a highly contagious, economically important viral disease of sheep and goats with high morbidity and mortality rates. A live attenuated PPR vaccine has been produced in Veterinary Serum and Vaccines Research Institute (VSVRI), Abbasia. It has been utilized to control PPR disease in endemic areas of the Arabian Gulf. In this study, the vaccine was tested for identity, sterility and safety. The identity of four batches of PPR vaccines was tested as per the Office International des Epizooties (OIE) guidelines using F and N-genes based RT-PCR technique to amplify fragments of F and N-proteins and using one step real time RT-PCR. All batches are also subjected to sterility tests for detection of bovine viral diarrhea virus as possible extraneous virus contaminant using ELISA and RT-PCR and detection of mycoplasma as possible bacterial contaminant using PCR technique. Safety testing was carried out in rodents to detect any nonspecific toxicity. By F-gene based RT-PCR, PPRV could be detected in only 2 samples out of 4samples (50%). By N-gene based RT-PCR, PPRV could be detected in all samples (100%). By real time RT-PCR, PPRV was detected in all the 4 samples. PPR vaccine batches were free from BVDV or mycoplasma contamination. Tested PPR vaccines were safe to be used. Results in this study showed that molecular techniques could be used for rapid, sensitive and specific evaluation of PPR vaccine including real time RT-PCR or Ngene based RT-PCR for identity testing. RT-PCR could be used as rapid and specific method for detection of BVDV as adventitious contaminant and PCR assay could be used for detection of bacterial contaminants as mycoplasma

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