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

ABSTRACT: 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 × 101 copies/?L 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, speci?c, and repeatable method for the quantitative detection of CaKoV and can, therefore, be used in epidemiological studies.

SUBMITTER: Wang Y

PROVIDER: S-EPMC7385718 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

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

Wang Yong Y Cui Yongqiu Y Li Yeqiu Y Zhang Da D Sun Jianfei J Guo Xu X Liu Guangqing G Jiang Shudong S Li Yongdong Y

Journal of virological methods 20200728

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<sup>1</sup> copies/μL with high specificity, stability, and repeatability. Moreover, ...[more]

PMID: 32730841

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