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.
Highlights
Pet dogs play an important role in humans’ daily lives
In this study, two new multiplex polymerase chain reactions (mPCRs) methods were developed for the detection of canine respiratory viruses (CRV, including canine adenovirus type 2 (CAV-2), canine distemper virus (CDV), canine influenza virus (CIV) and canine parainfluenza virus (CPIV)) and canine enteric viruses (CEV, including CAV-2, CanineCV, canine coronavirus (CCoV) and canine parvovirus (CPV)), and we indicated that the mPCR methods established here are simple and effective tools for detecting the viruses of interest
Premixed plasmids of CRV and CEV were used as templates to verify the mPCRs, and the products were visualized by 1.5% agarose gel electrophoresis
Summary
Pet dogs play an important role in humans’ daily lives. The emergence of new pathogens and the continuous circulation of common etiological agents in dog populations have complicated canine diseases [1]. Among these diseases, canine infectious respiratory diseases (CIRD) and viral enteritis pose notable threats to dog health. Multiplex PCR for detection of canine respiratory and enteric diseases viruses. Prevention and Control for Severe Clinical Animal Diseases (2017B030314142)
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