F Gene and N Gene based Reverse Transcription PCR for Molecular Characterization of Peste des Petits Ruminants Virus

DOI: 10.18805/ijar.B-3727    | Article Id: B-3727 | Page : 300-304
Citation :- F Gene and N Gene based Reverse Transcription PCR for Molecular Characterization of Peste des Petits Ruminants Virus.Indian Journal Of Animal Research.2020.(54):300-304
A.K. Pandey, D.M. Muglikar, P.P. Mhase, M.M. Pawade, S.N. Daphal, P.D. Pawar  prashant_vet@yahoo.co.in
Address : Department of Veterinary Microbiology, Krantisinh Nana Patil  College of Veterinary Science, Shirwal- 412 801, Maharashtra, India.
Submitted Date : 14-09-2018
Accepted Date : 17-12-2018

Abstract

Present investigation was undertaken to detect and characterize the PPR virus from different clinical tissue samples of 14 sheep and 17 goats with respiratory disease from Maharashtra, India. All animals were tested by Sandwich ELISA, of which 70.96% were found positive carrying high PPR virus inwhich 12 were sheep and 10 were goats respectively. For confirmation of PPR, molecular detection was performed with RT-PCR using F gene and N gene specific primers. Intestine samples accounted for highest percent positivity (75%) followed by blood (66.66%) and lymph node (62.5%) for presence of virus. Unusually higher positivity was observed in heart, liver and Kidney (60%, each) than normal predilection sites such as lungs (57.84%) and spleen (50%). While nasal swabs and blood were individually processed with F gene and N gene specific PCR, the triturated organs were pooled for processing into ‘Sample A’ comprised of heart, kidney, liver and intestine combined together and ‘Sample B’ comprising of lung, spleen and lymph nodes combined for the molecular detection of PPR yielding the products each of 372bp and 463bp sizes, respectively. Out of total 40 samples tested, 09/12 each from both sample A and B, while 02/10 nasal swabs resulted positive, respectively and all 06 blood samples remained negative. (F as well as N gene PCR methods were found best suitable for detection of PPR virus from tissue samples of small ruminants). 

Keywords

ELISA PPR RT-PCR Sequencing

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