Canine respiratory coronavirus (CRCoV) is commonly associated with canine kennel cough worldwide. Clinically infected dogs present coughing, sneezing, and nasal discharge. Severe infections may progress to pneumonia. Through serological surveys, CRCoV has been identified as a worldwide pathogen found in the respiratory tracts of dogs suffering from mild or severe respiratory disease. In this study, three dogs were obtained from a dog kennel. Over the previous 5 days, the dogs showed coughing, sneezing, and nasal discharge. To detect the etiologic pathogen, we performed multiplex RT-PCR (mRT-PCR) to amplify the genes encoding canine influenza virus matrix protein, canine distemper virus nucleocapsid protein, and CRCoV spike protein. Dot blotting was achieved with a CRCoV-specific probe. Nasal-secreting CRCoV was detected by the 442 bp CRCoV-positive PCR reaction in the nasal swabbing samples from dogs. Further, CRCoV-positive reactions by dot blot hybridization were detected in the nasal swabbing samples from dogs. In conclusion, we detected CRCoV in kenneled dogs with respiratory disease in Korea. Multiplex RT-PCR was able to detect successfully CRCoV infection in dogs. We suggest that mRT-PCR would be useful and effective for monitoring CRCoV infection in various kinds of dogs.

Introduction
 Materials and Methods
  Study animals and swabbing
  RNA extraction and cDNA synthesis
  Multiplex reverse transcription PCR
  Dot blot hybridization analysis
 Results
  Multiplex RT-PCR with nasal swabbing samples
  Dot blot hybridization for CRCoV
 Discussion
 Acknowledgements
 ORCID
 References

• Sunhwa Hong(Center for Animal Resource Development Research)
• Hak-Yong Lee(Huvet Co. Ltd., Wonkwang University, Iksan 570-749, Korea)
• Okjin Kim(Center for Animal Resource Development Research) Corresponding author