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

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).

A Necropsy based Investigation on Concurrent Infection of PPR and Emerging Opportunistic Bacterial Pathogens in Sheep and Goat

“…Out of 200 samples of nasal swab, tracheal swab, lungs and lung associated lymphnodes collected from 50 carcasses; twenty carcasses were found to be affected by PPR virus. PPRV antigen was detected equally i.e (100%) in nasal swab, tracheal swab, lung associated lymph nodes and lungs of eleven goats and nine sheep respectively (Pandey et al 2020). Similar study was conducted on different clinical samples of small ruminants (Mahajan et al 2012) for detection of PPR antigen which revealed lung and nasal swabs are more appropriate for detection of PPR antigen.…”

Epidemiology, Haemato-biochemical and Pathological Changes Related to Field Outbreaks of PPR in Small Ruminants in Odisha

“…All the samples were screened PCR positive (Fig 3) for PPRV. Molecular based confirmation for PPR virus is the most reliable and extensively used technique for diagnosis due to its high sensitivity and specificity (Kumar et al 2014;Pandey et al 2020).…”