Development of a new multiplex quantitative PCR for the detection of Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviae
Abstract Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviae are important porcine pathogens responsible for polyserositis, polyarthritis, meningitis, pneumonia, and septicemia causing significant economic losses in the swine industry. A new multiplex quantitative polymerase chai...
| Principais autores: | , , , , , , |
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| Formato: | Artigo |
| Idioma: | English |
| Publicado em: |
Wiley
2023-06-01
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| coleção: | MicrobiologyOpen |
| Assuntos: | |
| Acesso em linha: | https://doi.org/10.1002/mbo3.1353 |
| _version_ | 1827914585138528256 |
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| author | Simone Scherrer Sarah Schmitt Fenja Rademacher Peter Kuhnert Giovanni Ghielmetti Sophie Peterhans Roger Stephan |
| author_facet | Simone Scherrer Sarah Schmitt Fenja Rademacher Peter Kuhnert Giovanni Ghielmetti Sophie Peterhans Roger Stephan |
| author_sort | Simone Scherrer |
| collection | DOAJ |
| description | Abstract Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviae are important porcine pathogens responsible for polyserositis, polyarthritis, meningitis, pneumonia, and septicemia causing significant economic losses in the swine industry. A new multiplex quantitative polymerase chain reaction (qPCR) was designed on one hand for the detection of G. parasuis and the virulence marker vtaA to distinguish between highly virulent and non‐virulent strains. On the other hand, fluorescent probes were established for the detection and identification of both M. hyorhinis and M. hyosynoviae targeting 16S ribosomal RNA genes. The development of the qPCR was based on reference strains of 15 known serovars of G. parasuis, as well as on the type strains M. hyorhinis ATCC 17981T and M. hyosynoviae NCTC 10167T. The new qPCR was further evaluated using 21 G. parasuis, 26 M. hyorhinis, and 3 M. hyosynoviae field isolates. Moreover, a pilot study including different clinical specimens of 42 diseased pigs was performed. The specificity of the assay was 100% without cross‐reactivity or detection of other bacterial swine pathogens. The sensitivity of the new qPCR was demonstrated to be between 11–180 genome equivalents (GE) of DNA for M. hyosynoviae and M. hyorhinis, and 140–1200 GE for G. parasuis and vtaA. The cut‐off threshold cycle was found to be at 35. The developed sensitive and specific qPCR assay has the potential to become a useful molecular tool, which could be implemented in veterinary diagnostic laboratories for the detection and identification of G. parasuis, its virulence marker vtaA, M. hyorhinis, and M. hyosynoviae. |
| first_indexed | 2024-03-13T02:46:18Z |
| format | Article |
| id | doaj.art-4df64e9c8ae7466da68f91118eeb9cd1 |
| institution | Directory Open Access Journal |
| issn | 2045-8827 |
| language | English |
| last_indexed | 2024-03-13T02:46:18Z |
| publishDate | 2023-06-01 |
| publisher | Wiley |
| record_format | Article |
| series | MicrobiologyOpen |
| spelling | doaj.art-4df64e9c8ae7466da68f91118eeb9cd12023-06-28T18:00:37ZengWileyMicrobiologyOpen2045-88272023-06-01123n/an/a10.1002/mbo3.1353Development of a new multiplex quantitative PCR for the detection of Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviaeSimone Scherrer0Sarah Schmitt1Fenja Rademacher2Peter Kuhnert3Giovanni Ghielmetti4Sophie Peterhans5Roger Stephan6Section of Veterinary Bacteriology, Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich Zurich SwitzerlandSection of Veterinary Bacteriology, Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich Zurich SwitzerlandSection of Veterinary Bacteriology, Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich Zurich SwitzerlandInstitute of Veterinary Bacteriology, Vetsuisse Faculty University of Bern Bern SwitzerlandSection of Veterinary Bacteriology, Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich Zurich SwitzerlandSection of Veterinary Bacteriology, Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich Zurich SwitzerlandSection of Veterinary Bacteriology, Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich Zurich SwitzerlandAbstract Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviae are important porcine pathogens responsible for polyserositis, polyarthritis, meningitis, pneumonia, and septicemia causing significant economic losses in the swine industry. A new multiplex quantitative polymerase chain reaction (qPCR) was designed on one hand for the detection of G. parasuis and the virulence marker vtaA to distinguish between highly virulent and non‐virulent strains. On the other hand, fluorescent probes were established for the detection and identification of both M. hyorhinis and M. hyosynoviae targeting 16S ribosomal RNA genes. The development of the qPCR was based on reference strains of 15 known serovars of G. parasuis, as well as on the type strains M. hyorhinis ATCC 17981T and M. hyosynoviae NCTC 10167T. The new qPCR was further evaluated using 21 G. parasuis, 26 M. hyorhinis, and 3 M. hyosynoviae field isolates. Moreover, a pilot study including different clinical specimens of 42 diseased pigs was performed. The specificity of the assay was 100% without cross‐reactivity or detection of other bacterial swine pathogens. The sensitivity of the new qPCR was demonstrated to be between 11–180 genome equivalents (GE) of DNA for M. hyosynoviae and M. hyorhinis, and 140–1200 GE for G. parasuis and vtaA. The cut‐off threshold cycle was found to be at 35. The developed sensitive and specific qPCR assay has the potential to become a useful molecular tool, which could be implemented in veterinary diagnostic laboratories for the detection and identification of G. parasuis, its virulence marker vtaA, M. hyorhinis, and M. hyosynoviae.https://doi.org/10.1002/mbo3.1353Glaesserella parasuismultiplex qPCRMycoplasma hyorhinisMycoplasma hyosynoviaevtaA |
| spellingShingle | Simone Scherrer Sarah Schmitt Fenja Rademacher Peter Kuhnert Giovanni Ghielmetti Sophie Peterhans Roger Stephan Development of a new multiplex quantitative PCR for the detection of Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviae MicrobiologyOpen Glaesserella parasuis multiplex qPCR Mycoplasma hyorhinis Mycoplasma hyosynoviae vtaA |
| title | Development of a new multiplex quantitative PCR for the detection of Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviae |
| title_full | Development of a new multiplex quantitative PCR for the detection of Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviae |
| title_fullStr | Development of a new multiplex quantitative PCR for the detection of Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviae |
| title_full_unstemmed | Development of a new multiplex quantitative PCR for the detection of Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviae |
| title_short | Development of a new multiplex quantitative PCR for the detection of Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviae |
| title_sort | development of a new multiplex quantitative pcr for the detection of glaesserella parasuis mycoplasma hyorhinis and mycoplasma hyosynoviae |
| topic | Glaesserella parasuis multiplex qPCR Mycoplasma hyorhinis Mycoplasma hyosynoviae vtaA |
| url | https://doi.org/10.1002/mbo3.1353 |
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