Multiplex PCR methods for detection of several viruses associated with canine respiratory and enteric diseases

PLoS One. 2019 Mar 4;14(3):e0213295. doi: 10.1371/journal.pone.0213295. eCollection 2019.

Abstract

Viral respiratory and intestinal infections are the most common causes of canine viral illness. Infection with multiple pathogens occurs in many cases. Rapid diagnosis of these multiple infections is important for providing timely and effective treatment. To improve diagnosis, in this study, two new multiplex polymerase chain reactions (mPCRs) were developed for simultaneous detection of canine respiratory viruses (CRV) and canine enteric viruses (CEV) using two separate primer mixes. The viruses included canine adenovirus type 2 (CAV-2), canine distemper virus (CDV), canine influenza virus (CIV), canine parainfluenza virus (CPIV), canine circovirus (CanineCV), canine coronavirus (CCoV) and canine parvovirus (CPV). The sensitivity of the mPCR results showed that the detection limit of both mPCR methods was 1×104 viral copies. Twenty nasal swabs (NS) and 20 anal swabs (AS) collected from dogs with symptoms of respiratory disease or enteric disease were evaluated using the novel mPCR methods as a clinical test. The mPCR protocols, when applied to these respiratory specimens and intestinal samples, could detect 7 viruses simultaneously, allowing rapid investigation of CRV (CAV-2, CDV, CIV and CPIV) and CEV (CAV-2, CanineCV, CCoV and CPV) status and prompt evaluation of coinfection. Our study provides an effective and accurate tool for rapid differential diagnosis and epidemiological surveillance in dogs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Coronavirus Infections / veterinary*
  • Coronavirus Infections / virology
  • Coronavirus, Canine / classification
  • Coronavirus, Canine / genetics
  • Coronavirus, Canine / isolation & purification*
  • Dog Diseases / epidemiology*
  • Dog Diseases / virology
  • Dogs
  • Enterovirus / classification
  • Enterovirus / genetics
  • Enterovirus / isolation & purification*
  • Enterovirus Infections / veterinary*
  • Enterovirus Infections / virology
  • Multiplex Polymerase Chain Reaction / methods*
  • Respiration Disorders / veterinary*
  • Respiration Disorders / virology

Grants and funding

This work was supported in part by the National Natural Science Foundation of China (31672563, 31802204), The National Key Research and Development Program of China (2016YFD0501004), The Natural Science Foundation Guangdong province (2018B030311037, 2018A030313633), The Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases (2017B030314142).