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Development of ELISA Using Recombinant Proteins for the Diagnosis of Mycoplasma hyopneumoniae Infection

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Abstract

In order to develop a more sensitive and reliable method for detection of serum antibodies against Mycoplasma hyopneumoniae infection in pigs, six recombinant proteins of M. hyopneumoniae (P102, P95, P46, P97 like, Lppt, and hypothetical P987) were used for the standardization of an indirect enzyme-linked immunosorbent assay (ELISA). The proteins were evaluated against 50 sera of the specific pathogen-free and 50 sera of pigs with lesions suggestive of infection. The sensitivity was 88%, 86%, 78%, 74%, 66%, and 60% for the proteins P102, P95, P46, P97 like, Lppt, and hypothetical protein P987, respectively. Moreover, the proteins were used to establish the seroprevalence in two different commercial herds (254 sera pigs from farm considered free of M. hyopneumoniae and 246 from farm with clinical signs of enzootic pneumonia and positive serology for M. hyopneumoniae) and the positive rate was 65.2% for P95, 54.6% for P102, 40.2% for P46, 37.2% for P97 like, 17.4% for the hypothetical P987, and 14% for Lppt protein. In addition, the ELISA with six recombinant proteins was compared to commercial HerdCheck kit using 118 random pig sera samples and the results showed that ELISA with recombinant proteins were more sensitive than the commercial test. These data show that the recombinant proteins P95 and P102 are potential targets to be used in diagnostic tests to detect antibodies against M. hyopneumoniae. Although more studies are necessary, this study provides insights that these recombinant proteins can be useful in epidemiological investigations and as potential biomarkers in differentiating infected animals from those vaccinated.

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Acknowledgements

We are grateful to Davi Barcellos and Andrea Panzardi for providing convalescent pig sera and Michele Ribeiro dos Santos for technical assistance.

Funding

This study was supported by the Brazilian National Research Council (CNPq grants 578614/2008–1 and 303148/2009–8), Fundação de Apoio ao Desenvolvimento do Ensino, Ciência e Tecnologia do Estado de Mato Grosso do Sul (FUNDECT grants 092/2015), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).

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Author notes

  1. Simone Simionatto and Silvana Beutinger Marchioro have contributed equally to this work.

Authors and Affiliations

  1. Laboratório de Pesquisa em Ciências da Saúde, Universidade Federal da Grande Dourados, Dourados, MS, Brazil

    Simone Simionatto, Silvana Beutinger Marchioro & Marcelo dos Santos Barbosa

  2. Laboratório de Imunologia e Biologia Molecular, Instituto de Ciências da Saúde, Universidade Federal da Bahia, Salvador, BA, Brazil

    Silvana Beutinger Marchioro

  3. Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil

    Vanessa Galli, Clarice Brink Brum, Sergio Jorge & Odir Antonio Dellagostin

  4. Programa de Pós Graduação em Epidemiologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil

    Clarice Brink Brum

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  1. Simone Simionatto
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  2. Silvana Beutinger Marchioro
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Contributions

Conceptualization, OAD, SBM, and SS; Methodology, CBB, MSB, SJ, and VG; Formal analysis and investigation, CBB, MSB, OAD, SBM, SJ, SS, and VG; Writing - original draft preparation, CBB, MSB, OAD, SBM, SJ, SS, and VG; Writing -review and editing, SBM, SS, and OAD; Funding acquisition, SBM, and SS; Supervision, SBM, and SS. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Silvana Beutinger Marchioro.

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None of the authors has a financial or personal relationship with other people or organizations that could inappropriately influence or bias the content of the paper.

Ethics Approval

All animal procedures were performed at the animal facility of the Federal University of Pelotas (UFPel) and approved by the Ethics Committee for Animal Experimentation (CEEA) of UFPel under protocol number 5614. The CEEA of UFPel is accredited by the Brazilian National Council for Animal Experimentation Control (CONCEA).

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Simionatto, S., Marchioro, S.B., dos Santos Barbosa, M. et al. Development of ELISA Using Recombinant Proteins for the Diagnosis of Mycoplasma hyopneumoniae Infection. Indian J Microbiol 62, 88–95 (2022). https://doi.org/10.1007/s12088-021-00981-z

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  • DOI: https://doi.org/10.1007/s12088-021-00981-z

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