Nested polymerase chain reaction assay for detection of Porcine Astrovirus in pig population

AAKRITI  PATHANIA; ADARSH  MISHRA; VISHAL  MAHAJAN; YASHPAL SINGH  MALIK

doi:10.56093/ijans.v95i11.166592

Authors

AAKRITI PATHANIA College of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab, India
ADARSH MISHRA College of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab, India
VISHAL MAHAJAN 2Animal Disease Research Centre, College of Veterinary Science, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab, India
YASHPAL SINGH MALIK College of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab, India

https://doi.org/10.56093/ijans.v95i11.166592

Keywords:

Porcine, Gastroenteritis, Astrovirus, Nested PCR, Specificity, Sensitivity

Abstract

Porcine Astrovirus (PAstV) is a positive-sense single-stranded RNA virus, responsible for gastrointestinal diseases in swine populations across the globe. There are sparse reports regarding diagnostic platforms for the detection of PAstV in the porcine population. The present study reported a nested PCR (nPCR) assay for the identification of PAstV in faecal specimens of pigs in Punjab, India. The technique was developed using outer and inner sets of primers targeting the RNA-dependent RNA polymerase (RdRp) gene of the virus. The nPCR was standardized and optimized, including sensitivity and specificity. The sensitivity of the assay was found to be 225 femtograms. The technique was specific and did not amplify with certain other viruses i.e., Porcine Kobuvirus (PKV) and Bovine Rotavirus (BoRV). The applicability of the nPCR assay was further assessed on faecal samples (n=50) collected from field conditions comprising diarrhoeic (n=39) and non-diarrhoeic (n=11) animals. The nPCR detected only 12 [diarrhoeic (n=8) & non-diarrhoeic (n=4)] out of 50 samples tested as positive. This work demonstrated that nPCR assay can be effectively used as a rapid, specific, and sensitive method for routine molecular screening and/or diagnosis of PAstV in swine population.

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