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ORIGINAL RESEARCH article

Front. Cell. Infect. Microbiol.
Sec. Veterinary and Zoonotic Infection
Volume 14 - 2024 | doi: 10.3389/fcimb.2024.1448480
This article is part of the Research Topic Unveiling Host-Pathogen Interactions: Insights into Animal Cellular Immunity and Novel Diagnostics View all 7 articles

Development and application of quadruplex real time quantitative PCR method for differentiation of Muscovy duck parvovirus, Goose parvovirus, Duck circovirus, and Duck adenovirus 3

Provisionally accepted
He Zhang He Zhang 1*Haojie Wang Haojie Wang Haojie Wang Haojie Wang 1Jianxing Chen Jianxing Chen 1Tong-Qing An Tong-Qing An 1Hongyan Chen Hongyan Chen 1Yue Wang Yue Wang 1LIANGQUAN ZHU LIANGQUAN ZHU 2Changqing Yu Changqing Yu 3*Changyou Xia Changyou Xia 1
  • 1 Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China
  • 2 China Institute of Veterinary Drug Control, Beijing, Beijing Municipality, China
  • 3 Yibin Vocational and Technical College, Yibin, Sichuan Province, China

The final, formatted version of the article will be published soon.

    Muscovy duck parvovirus (MDPV), Goose parvovirus (GPV), Duck circovirus, (DuCV) and Duck adenovirus 3 (DAdV-3) are important pathogens that cause high morbidity and mortality in ducks, causing huge economic loss for the duck industry. The present study, a quadruplex one-step real time quantitative PCR method for the detection of MDPV, GPV, DuCV, and DAdV-3 was developed. The results showed that assay had no cross-reactivity with other poultry pathogens [Duck plague virus (DPV), Duck tembusu virus (DTMUV), H6 avian influenza virus (H6 AIV), New duck reovirus (NDRV), Newcastle disease virus (NDV), H4 avian influenza virus (H4 AIV), Escherichia coli (E. coli), Muscovy duck reovirus (MDRV), Egg drop syndrome virus (EDSV), Pasteurella multocida (P. multocida)]. The sensitivity result showed that the limits of detection for MDPV, GPV, DuCV, and DAdV-3 were 10, 10, 1 and 10 copies/µl, respectively; The coefficients of variation intra-and inter-method was 1-2%; The range of linear (10 9 to 10 2 10 3 copies/µL) demonstrated the R 2 values for MDPV, GPV, DuCV, and DAdV-3 as 0.9975, 0.998, 0.9964, and 0.996, respectively. The quadruplex real time quantitative PCR method efficiency was 90.30%, 101.10%, 90.72%, and 90.57% for MDPV, GPV, DuCV, and DAdV-3, respectively. 396 clinical specimens collected in some duck sausages from June 2022 to July 2023 were simultaneously detected using the established quadruplex real time quantitative PCR method and the reported assays. The detection rates for MDPV, GPV, DuCV, and DAdV-3 were 8.33% (33/396), 17.93% (71/396), 33.58% (133/396), and 29.04% (115/396), respectively. The agreement between these assays was greater than 99.56%. The developed quadruplex real-time quantitative PCR assay can accurately detect these four viruses infecting ducks, providing a rapid, sensitive, specific and accurate technique for clinical testing.

    Keywords: muscovy duck parvovirus (MDPV), Goose parvovirus (GPV), Duck circovirus, (DuCV), duck adenovirus 3 (DAdV-3), Quadruplex, qPCR

    Received: 13 Jun 2024; Accepted: 23 Jul 2024.

    Copyright: © 2024 Zhang, Haojie Wang, Chen, An, Chen, Wang, ZHU, Yu and Xia. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence:
    He Zhang, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China
    Changqing Yu, Yibin Vocational and Technical College, Yibin, Sichuan Province, China

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.