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ORIGINAL RESEARCH article

Front. Vet. Sci.
Sec. Veterinary Experimental and Diagnostic Pathology
Volume 11 - 2024 | doi: 10.3389/fvets.2024.1440124

Establishment of ELISA method for canine adenovirus type 1

Provisionally accepted
Ben Wang Ben Wang 1Jinfeng Xu Jinfeng Xu 1Hui Zhang Hui Zhang 2Shizhen Lian Shizhen Lian 2Yichang Duan Yichang Duan 3Hongling Zhang Hongling Zhang 1Wei Hou Wei Hou 2Yin B. Shuang Yin B. Shuang 1Yanzhu Zhu Yanzhu Zhu 1*
  • 1 Jilin Agricultural Science and Technology College, Jilin, Jilin, China
  • 2 Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, Jilin Province, China
  • 3 Heilongjiang Bayi Agricultural University, Daqing, Heilongjiang Province, China

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

    Canine adenovirus (CAdV) had a high prevalence in fox populations and induced fox encephalitis. No ELISA kits specifically for CAdV-1 antigen had been commercialized for foxes in China. It is crucial to develop a rapid and accurate ELISA method for detecting of CAdV-1. The monoclonal antibodies (mAbs, IgG1A) and HRP-labeled polyclonal antibodies (pAbs) were used to establish the ELISA method in this experiment. The results showed that the optimal concentration and coating time for the mAbs (IgG1A) were 2.15 μg/mL and overnight at 4 ℃, respectively. The dilution ratio of the HRP-labeled pAbs was 1:2000.Five percent skimmed milk was selected as the blocking agent. The optimal incubation times for blocking, CAdV-1, and HRP-labeled pAbs were all 1 hour. The cut-off value for negative rectal swab was determined to be 0.366±0.032. The maximum dilution ratio was 100 TCID50/mL. The ELISA method was positive to CAdV-1, and that was negative to CAdV-2, Canine Parvovirus (CPV) and Canine Distempervirus (CDV).The ELISA method showed good repeatability, sensitivity, and specificity. Compared with RT-PCR, the sensitivity, specificity, and coincidence rates of the ELISA method were 93.75%, 90.9%, and 92.86%, respectively. These results indicate that the established ELISA method can be used for the large-scale screening and epidemiology surveillance of CAdV-1 in foxes.

    Keywords: canine adenovirus type 1, ELISA method, Sensitivity, specificity, Coincidence rates

    Received: 29 May 2024; Accepted: 09 Aug 2024.

    Copyright: © 2024 Wang, Xu, Zhang, Lian, Duan, Zhang, Hou, Shuang and Zhu. 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: Yanzhu Zhu, Jilin Agricultural Science and Technology College, Jilin, Jilin, China

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