Development of a multiplex RT-PCR for the detection of major diarrhoeal viruses in pig herds in China

Transbound Emerg Dis. 2020 Mar;67(2):678-685. doi: 10.1111/tbed.13385. Epub 2019 Oct 28.

Abstract

The major enteric RNA viruses in pigs include porcine epidemic diarrhoea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine rotavirus A (PRV-A), porcine kobuvirus (PKV), porcine sapovirus (PSaV) and porcine deltacoronavirus (PDCoV). For differential diagnosis, a multiplex RT-PCR method was established on the basis of the N genes of TGEV, PEDV and PDCoV, the VP7 gene of PRV-A, and the polyprotein genes of PKV and PSaV. This multiplex RT-PCR could specifically detect TGEV, PEDV, PDCoV, PRV-A, PKV and PSaV without cross-reaction to any other major viruses circulating in Chinese pig farms. The limit of detection of this method was as low as 100 -101 ng cDNA of each virus. A total of 398 swine faecal samples collected from nine provinces of China between October 2015 and April 2017 were analysed by this established multiplex RT-PCR. The results demonstrated that PDCoV (144/398), PSaV (114/398), PEDV (78/398) and PRV-A (70/398) were the main pathogens, but TGEV was not found in the pig herds in China. In addition, dual infections, for example, PDCoV + PSaV, PDCoV + PRV-A, PRA-V + PSaV and PEDV + PDCoV, and triple infections, for example, PDCoV + PRV-A + PSaV and PEDV + PDCoV + PKV, were found among the collected samples. The multiplex RT-PCR provided a valuable tool for the differential diagnosis of swine enteric viruses circulating in Chinese pig farms and will facilitate the prevention and control of swine diarrhoea in China.

Keywords: diarrhoea; multiplex RT-PCR; swine enteric viruses.

MeSH terms

  • Animals
  • China / epidemiology
  • Coronavirus Infections / diagnosis
  • Coronavirus Infections / epidemiology
  • Coronavirus Infections / veterinary*
  • Coronavirus Infections / virology
  • Diagnosis, Differential
  • Diarrhea / diagnosis
  • Diarrhea / prevention & control
  • Diarrhea / veterinary*
  • Diarrhea / virology
  • Feces / virology
  • Gastroenteritis, Transmissible, of Swine / virology
  • Kobuvirus / genetics
  • Kobuvirus / isolation & purification
  • Multiplex Polymerase Chain Reaction / veterinary*
  • Porcine epidemic diarrhea virus / genetics
  • Porcine epidemic diarrhea virus / isolation & purification
  • Reverse Transcriptase Polymerase Chain Reaction / veterinary
  • Rotavirus / genetics
  • Rotavirus / isolation & purification
  • Sensitivity and Specificity
  • Swine
  • Swine Diseases / diagnosis*
  • Swine Diseases / epidemiology
  • Swine Diseases / virology