Development of an SYBR Green I-based real-time PCR assay for the rapid detection of canine kobuvirus

Wang, Yong; Cui, Yongqiu; Li, Yeqiu; Zhang, Da; Sun, Jianfei; Guo, Xu; Liu, Guangqing; Jiang, Shudong; Li, Yongdong

Canine kobuvirus (CaKoV) is a causative agent of gastroenteritis in dogs. Rapid detection of CaKoV is important for preventing and controlling this condition. In this study, an SYBR Green I-based quantitative real-time PCR assay was established for CaKoV detection. Specific primers targeting a highly conserved region of the CaKoV 3D gene were developed. After optimization, the method detected a minimum of 1 × 10(1) copies/µL with high specificity, stability, and repeatability. Moreover, the entire process only required approximately 1.5 h for completion. Our results were supported by those obtained for clinical samples, in which our developed method was successfully applied. The newly established real-time PCR is a rapid, sensitive, specific, and repeatable method for the quantitative detection of CaKoV and can, therefore, be used in epidemiological studies.