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ORIGINAL RESEARCH article

Front. Vet. Sci.
Sec. Veterinary Infectious Diseases
Volume 12 - 2025 | doi: 10.3389/fvets.2025.1532600

Development and Evaluation of Optimized PCR and Indirect ELISA for the Detection of Morganella morganii in Dairy Cows

Provisionally accepted
Meihua Zhang Meihua Zhang 1Jiayi Li Jiayi Li 1Jianfeng Xue Jianfeng Xue 2Huiling Xu Huiling Xu 1Muzi Li Muzi Li 1Yibo Xia Yibo Xia 1Changxi Qi Changxi Qi 1Pu Zhang Pu Zhang 2*Yongxia Liu Yongxia Liu 1*Jianzhu Liu Jianzhu Liu 1*
  • 1 Shandong Agricultural University, Taian, China
  • 2 The Affiliated Hospital of Qingdao University, Qingdao, Shandong Province, China

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

    Morganella morganii (M. morganii) is a Gram-negative opportunistic pathogen, whose increasing virulence and antibiotic resistance negatively impact dairy cow health and productivity, raising concerns in livestock health management. To mitigate this risk, rapid and reliable diagnostic methods for detection are essential. Currently, detection methods for M. morganii are underdeveloped, prompting us to develop both pathogenic and serological detection methods, including an optimized PCR technique and an indirect enzyme-linked immunosorbent assay (I-ELISA). The optimized PCR method uses bacterial suspension as a template, eliminating DNA extraction and allowing direct detection of fecal samples, improving efficiency. By optimizing primer concentration and adjusting annealing temperature, primer dimer formation was minimized, ensuring high specificity. The method has high sensitivity, with a minimum detection concentration of 0.2 CFU/μL. In clinical applications, 771 fecal and nasal fluid samples from dairy farms across five regions were tested, with a positivity rate of 1.4%. The I-ELISA method, developed using M. morganii lipoprotein (LPP) antigen, improved specificity through optimization of antigen coating, blocking conditions, and antibody dilution. Its high stability and reproducibility were confirmed through intra- and inter-assay tests. In testing 476 dairy cow serum samples, the I-ELISA positivity rate was 5.9%. Collectively, the PCR and I-ELISA detection methods provide practical solutions for early clinical diagnosis, facilitating timely interventions against M. morganii infections.

    Keywords: enzyme linked immunosorbent assay, Epidemiological investigation, lipoprotein, Morganella morganii, Polymerase Chain Reaction

    Received: 22 Nov 2024; Accepted: 02 Jan 2025.

    Copyright: © 2025 Zhang, Li, Xue, Xu, Li, Xia, Qi, Zhang, Liu and Liu. 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:
    Pu Zhang, The Affiliated Hospital of Qingdao University, Qingdao, 266000, Shandong Province, China
    Yongxia Liu, Shandong Agricultural University, Taian, China
    Jianzhu Liu, Shandong Agricultural University, Taian, 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.