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A simplified PCR assay for fast and easy mycoplasma mastitis screening in dairy cattle
A simplified polymerase chain reaction (PCR) assay was developed for fast and easy screening of mycoplasma mastitis in dairy cattle. Species of major mycoplasma strains [Mycoplasma (M.) bovis, M. arginini, M. bovigenitalium, M. californicum, M. bovirhinis, M. alkalescens and M. canadense] in culture...
Autores principales: | , , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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The Korean Society of Veterinary Science
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3104175/ https://www.ncbi.nlm.nih.gov/pubmed/21586880 http://dx.doi.org/10.4142/jvs.2011.12.2.191 |
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author | Higuchi, Hidetoshi Iwano, Hidetomo Kawai, Kazuhiro Ohta, Takehiro Obayashi, Tetsu Hirose, Kazuhiko Ito, Nobuhiko Yokota, Hiroshi Tamura, Yutaka Nagahata, Hajime |
author_facet | Higuchi, Hidetoshi Iwano, Hidetomo Kawai, Kazuhiro Ohta, Takehiro Obayashi, Tetsu Hirose, Kazuhiko Ito, Nobuhiko Yokota, Hiroshi Tamura, Yutaka Nagahata, Hajime |
author_sort | Higuchi, Hidetoshi |
collection | PubMed |
description | A simplified polymerase chain reaction (PCR) assay was developed for fast and easy screening of mycoplasma mastitis in dairy cattle. Species of major mycoplasma strains [Mycoplasma (M.) bovis, M. arginini, M. bovigenitalium, M. californicum, M. bovirhinis, M. alkalescens and M. canadense] in cultured milk samples were detected by this simplified PCR-based method as well as a standard PCR technique. The minimum concentration limit for detecting mycoplasma by the simplified PCR was estimated to be about 2.5 × 10(3) cfu/mL and was similar to that of the standard PCR. We compared the specificity and sensitivity of the simplified PCR to those of a culture method. Out of 1,685 milk samples cultured in mycoplasma broth, the simplified PCR detected Mycoplasma DNA in 152 that were also positive according to the culture assay. The sensitivity and specificity of the simplified PCR were 98.7% and 99.7%, respectively, for detecting mycoplasma in those cultures. The results obtained by the simplified PCR were consistent with ones from standard PCR. This newly developed simplified PCR, which does not require DNA purification, can analyze about 300 cultured samples within 3 h. The results from our study suggest that the simplified PCR can be used for mycoplasma mastitis screening in large-scale dairy farms. |
format | Text |
id | pubmed-3104175 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | The Korean Society of Veterinary Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-31041752011-06-09 A simplified PCR assay for fast and easy mycoplasma mastitis screening in dairy cattle Higuchi, Hidetoshi Iwano, Hidetomo Kawai, Kazuhiro Ohta, Takehiro Obayashi, Tetsu Hirose, Kazuhiko Ito, Nobuhiko Yokota, Hiroshi Tamura, Yutaka Nagahata, Hajime J Vet Sci Short Communication A simplified polymerase chain reaction (PCR) assay was developed for fast and easy screening of mycoplasma mastitis in dairy cattle. Species of major mycoplasma strains [Mycoplasma (M.) bovis, M. arginini, M. bovigenitalium, M. californicum, M. bovirhinis, M. alkalescens and M. canadense] in cultured milk samples were detected by this simplified PCR-based method as well as a standard PCR technique. The minimum concentration limit for detecting mycoplasma by the simplified PCR was estimated to be about 2.5 × 10(3) cfu/mL and was similar to that of the standard PCR. We compared the specificity and sensitivity of the simplified PCR to those of a culture method. Out of 1,685 milk samples cultured in mycoplasma broth, the simplified PCR detected Mycoplasma DNA in 152 that were also positive according to the culture assay. The sensitivity and specificity of the simplified PCR were 98.7% and 99.7%, respectively, for detecting mycoplasma in those cultures. The results obtained by the simplified PCR were consistent with ones from standard PCR. This newly developed simplified PCR, which does not require DNA purification, can analyze about 300 cultured samples within 3 h. The results from our study suggest that the simplified PCR can be used for mycoplasma mastitis screening in large-scale dairy farms. The Korean Society of Veterinary Science 2011-06 2011-05-19 /pmc/articles/PMC3104175/ /pubmed/21586880 http://dx.doi.org/10.4142/jvs.2011.12.2.191 Text en Copyright © 2011 The Korean Society of Veterinary Science https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0 (https://creativecommons.org/licenses/by-nc/4.0/) ) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Short Communication Higuchi, Hidetoshi Iwano, Hidetomo Kawai, Kazuhiro Ohta, Takehiro Obayashi, Tetsu Hirose, Kazuhiko Ito, Nobuhiko Yokota, Hiroshi Tamura, Yutaka Nagahata, Hajime A simplified PCR assay for fast and easy mycoplasma mastitis screening in dairy cattle |
title | A simplified PCR assay for fast and easy mycoplasma mastitis screening in dairy cattle |
title_full | A simplified PCR assay for fast and easy mycoplasma mastitis screening in dairy cattle |
title_fullStr | A simplified PCR assay for fast and easy mycoplasma mastitis screening in dairy cattle |
title_full_unstemmed | A simplified PCR assay for fast and easy mycoplasma mastitis screening in dairy cattle |
title_short | A simplified PCR assay for fast and easy mycoplasma mastitis screening in dairy cattle |
title_sort | simplified pcr assay for fast and easy mycoplasma mastitis screening in dairy cattle |
topic | Short Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3104175/ https://www.ncbi.nlm.nih.gov/pubmed/21586880 http://dx.doi.org/10.4142/jvs.2011.12.2.191 |
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