Abstract
Duck circovirus (DuCV) is widespread across the world and causes feather disorders in young ducks. It was identified as the causative pathogen of duck beak atrophy and dwarfism syndrome and primary sclerosing cholangitis. In this study, we aimed to establish a TaqMan-based real-time PCR assay to detect DuCV. The primers and probe were designed based on the conserved region of the DuCV Rep gene. After optimizing the reaction conditions, the minimum virus detection limit of the designed PCR technique was 39.4 copies/μL, 100 times that of conventional PCR (cPCR). No cross-reaction with six other common duck viruses was observed. The intra- and inter-assay variations were less than 1%. The detection rate of DuCV-positive clinical samples using TaqMan-based real-time PCR was higher than that using SYBR Green-based real-time PCR and cPCR. Collectively, these results showed that the established TaqMan-based real-time PCR detected DuCV with high sensitivity and specificity, and significant repeatability, making it suitable for clinical use. Hence, it may be used as a novel tool for the diagnosis and epidemiological investigation of DuCV.
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We would like to thank Editage (www.editage.cn) for English language editing.
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This work was supported financially by Ningbo Health Branding Subject Fund (No. ppxk2018-10).
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Da Zhang and Junhuang Wu conceived of the study, carried out the experiment and drafted the manuscript, contributed equally to this work. Jianfei Sun and Caixia Bai participated in the data collection and analysis. Fazhi Xu and Pengfei Duan participated in statistical analysis. Yong Wang conceived of the study, revising the manuscript critically. All authors have read and approved the final manuscript.
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Zhang, D., Wu, J., Sun, J. et al. Establishment of TaqMan-based real-time PCR assay for rapid detection of duck circovirus. 3 Biotech 11, 470 (2021). https://doi.org/10.1007/s13205-021-03021-1
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DOI: https://doi.org/10.1007/s13205-021-03021-1