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Evaluation of a novel multiplex qPCR method for rapid detection and quantification of pathogens associated with calf diarrhoea
AIMS: Diarrhoea is a common health problem in calves and a main reason for use of antimicrobials. It is associated with several bacterial, viral and parasitic pathogens, most of which are commonly present in healthy animals. Methods, which quantify the causative agents, may therefore improve confide...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9796748/ https://www.ncbi.nlm.nih.gov/pubmed/35858716 http://dx.doi.org/10.1111/jam.15722 |
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author | Pansri, Potjamas Svensmark, Birgitta Liu, Gang Thamsborg, Stig Milan Kudirkiene, Egle Nielsen, Henrik Vedel Goecke, Nicole Bakkegård Olsen, John Elmerdahl |
author_facet | Pansri, Potjamas Svensmark, Birgitta Liu, Gang Thamsborg, Stig Milan Kudirkiene, Egle Nielsen, Henrik Vedel Goecke, Nicole Bakkegård Olsen, John Elmerdahl |
author_sort | Pansri, Potjamas |
collection | PubMed |
description | AIMS: Diarrhoea is a common health problem in calves and a main reason for use of antimicrobials. It is associated with several bacterial, viral and parasitic pathogens, most of which are commonly present in healthy animals. Methods, which quantify the causative agents, may therefore improve confidence in associating a pathogen to the disease. This study evaluated a novel commercially available, multiplex quantitative polymerase chain reaction (qPCR) assay (Enterit4Calves) for detection and quantification of pathogens associated with calf‐diarrhoea. METHODS AND RESULTS: Performance of the method was first evaluated under laboratory conditions. Then it was compared with current routine methods for detection of pathogens in faecal samples from 65 calves with diarrhoea and in 30 spiked faecal samples. The qPCR efficiencies were between 84%–103% and detection limits of 100–1000 copies of nucleic acids per sample were observed. Correct identification was obtained on 42 strains of cultured target bacteria, with only one false positive reaction from 135 nontarget bacteria. Kappa values for agreement between the novel assay and current routine methods varied between 0.38 and 0.83. CONCLUSION: The novel qPCR method showed good performance under laboratory conditions and a fair to good agreement with current routine methods when used for testing of field samples. SIGNIFICANCE AND IMPACT OF STUDY: In addition to having fair to good detection abilities, the novel qPCR method allowed quantification of pathogens. In the future, use of quantification may improve diagnosis and hence treatment of calf diarrhoea. |
format | Online Article Text |
id | pubmed-9796748 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-97967482023-01-04 Evaluation of a novel multiplex qPCR method for rapid detection and quantification of pathogens associated with calf diarrhoea Pansri, Potjamas Svensmark, Birgitta Liu, Gang Thamsborg, Stig Milan Kudirkiene, Egle Nielsen, Henrik Vedel Goecke, Nicole Bakkegård Olsen, John Elmerdahl J Appl Microbiol Original Articles AIMS: Diarrhoea is a common health problem in calves and a main reason for use of antimicrobials. It is associated with several bacterial, viral and parasitic pathogens, most of which are commonly present in healthy animals. Methods, which quantify the causative agents, may therefore improve confidence in associating a pathogen to the disease. This study evaluated a novel commercially available, multiplex quantitative polymerase chain reaction (qPCR) assay (Enterit4Calves) for detection and quantification of pathogens associated with calf‐diarrhoea. METHODS AND RESULTS: Performance of the method was first evaluated under laboratory conditions. Then it was compared with current routine methods for detection of pathogens in faecal samples from 65 calves with diarrhoea and in 30 spiked faecal samples. The qPCR efficiencies were between 84%–103% and detection limits of 100–1000 copies of nucleic acids per sample were observed. Correct identification was obtained on 42 strains of cultured target bacteria, with only one false positive reaction from 135 nontarget bacteria. Kappa values for agreement between the novel assay and current routine methods varied between 0.38 and 0.83. CONCLUSION: The novel qPCR method showed good performance under laboratory conditions and a fair to good agreement with current routine methods when used for testing of field samples. SIGNIFICANCE AND IMPACT OF STUDY: In addition to having fair to good detection abilities, the novel qPCR method allowed quantification of pathogens. In the future, use of quantification may improve diagnosis and hence treatment of calf diarrhoea. John Wiley and Sons Inc. 2022-07-31 2022-10 /pmc/articles/PMC9796748/ /pubmed/35858716 http://dx.doi.org/10.1111/jam.15722 Text en © 2022 The Authors. Journal of Applied Microbiology published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Original Articles Pansri, Potjamas Svensmark, Birgitta Liu, Gang Thamsborg, Stig Milan Kudirkiene, Egle Nielsen, Henrik Vedel Goecke, Nicole Bakkegård Olsen, John Elmerdahl Evaluation of a novel multiplex qPCR method for rapid detection and quantification of pathogens associated with calf diarrhoea |
title | Evaluation of a novel multiplex qPCR method for rapid detection and quantification of pathogens associated with calf diarrhoea |
title_full | Evaluation of a novel multiplex qPCR method for rapid detection and quantification of pathogens associated with calf diarrhoea |
title_fullStr | Evaluation of a novel multiplex qPCR method for rapid detection and quantification of pathogens associated with calf diarrhoea |
title_full_unstemmed | Evaluation of a novel multiplex qPCR method for rapid detection and quantification of pathogens associated with calf diarrhoea |
title_short | Evaluation of a novel multiplex qPCR method for rapid detection and quantification of pathogens associated with calf diarrhoea |
title_sort | evaluation of a novel multiplex qpcr method for rapid detection and quantification of pathogens associated with calf diarrhoea |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9796748/ https://www.ncbi.nlm.nih.gov/pubmed/35858716 http://dx.doi.org/10.1111/jam.15722 |
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