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A high-throughput qPCR system for simultaneous quantitative detection of dairy Lactococcus lactis and Leuconostoc bacteriophages
Simultaneous quantitative detection of Lactococcus (Lc.) lactis and Leuconostoc species bacteriophages (phages) has not been reported in dairies using undefined mixed-strain DL-starters, probably due to the lack of applicable methods. We optimized a high-throughput qPCR system that allows simultaneo...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5365131/ https://www.ncbi.nlm.nih.gov/pubmed/28339484 http://dx.doi.org/10.1371/journal.pone.0174223 |
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author | Muhammed, Musemma K. Krych, Lukasz Nielsen, Dennis S. Vogensen, Finn K. |
author_facet | Muhammed, Musemma K. Krych, Lukasz Nielsen, Dennis S. Vogensen, Finn K. |
author_sort | Muhammed, Musemma K. |
collection | PubMed |
description | Simultaneous quantitative detection of Lactococcus (Lc.) lactis and Leuconostoc species bacteriophages (phages) has not been reported in dairies using undefined mixed-strain DL-starters, probably due to the lack of applicable methods. We optimized a high-throughput qPCR system that allows simultaneous quantitative detection of Lc. lactis 936 (now SK1virus), P335, c2 (now C2virus) and Leuconostoc phage groups. Component assays are designed to have high efficiencies and nearly the same dynamic detection ranges, i.e., from ~1.1 x 10(5) to ~1.1 x 10(1) phage genomes per reaction, which corresponds to ~9 x 10(7) to ~9 x 10(3) phage particles mL(-1) without any additional up-concentrating steps. The amplification efficiencies of the corresponding assays were 100.1±2.6, 98.7±2.3, 101.0±2.3 and 96.2±6.2. The qPCR system was tested on samples obtained from a dairy plant that employed traditional mother-bulk-cheese vat system. High levels of 936 and P335 phages were detected in the mother culture and the bulk starter, but also in the whey samples. Low levels of phages were detected in the cheese milk samples. |
format | Online Article Text |
id | pubmed-5365131 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-53651312017-04-06 A high-throughput qPCR system for simultaneous quantitative detection of dairy Lactococcus lactis and Leuconostoc bacteriophages Muhammed, Musemma K. Krych, Lukasz Nielsen, Dennis S. Vogensen, Finn K. PLoS One Research Article Simultaneous quantitative detection of Lactococcus (Lc.) lactis and Leuconostoc species bacteriophages (phages) has not been reported in dairies using undefined mixed-strain DL-starters, probably due to the lack of applicable methods. We optimized a high-throughput qPCR system that allows simultaneous quantitative detection of Lc. lactis 936 (now SK1virus), P335, c2 (now C2virus) and Leuconostoc phage groups. Component assays are designed to have high efficiencies and nearly the same dynamic detection ranges, i.e., from ~1.1 x 10(5) to ~1.1 x 10(1) phage genomes per reaction, which corresponds to ~9 x 10(7) to ~9 x 10(3) phage particles mL(-1) without any additional up-concentrating steps. The amplification efficiencies of the corresponding assays were 100.1±2.6, 98.7±2.3, 101.0±2.3 and 96.2±6.2. The qPCR system was tested on samples obtained from a dairy plant that employed traditional mother-bulk-cheese vat system. High levels of 936 and P335 phages were detected in the mother culture and the bulk starter, but also in the whey samples. Low levels of phages were detected in the cheese milk samples. Public Library of Science 2017-03-24 /pmc/articles/PMC5365131/ /pubmed/28339484 http://dx.doi.org/10.1371/journal.pone.0174223 Text en © 2017 Muhammed et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Muhammed, Musemma K. Krych, Lukasz Nielsen, Dennis S. Vogensen, Finn K. A high-throughput qPCR system for simultaneous quantitative detection of dairy Lactococcus lactis and Leuconostoc bacteriophages |
title | A high-throughput qPCR system for simultaneous quantitative detection of dairy Lactococcus lactis and Leuconostoc bacteriophages |
title_full | A high-throughput qPCR system for simultaneous quantitative detection of dairy Lactococcus lactis and Leuconostoc bacteriophages |
title_fullStr | A high-throughput qPCR system for simultaneous quantitative detection of dairy Lactococcus lactis and Leuconostoc bacteriophages |
title_full_unstemmed | A high-throughput qPCR system for simultaneous quantitative detection of dairy Lactococcus lactis and Leuconostoc bacteriophages |
title_short | A high-throughput qPCR system for simultaneous quantitative detection of dairy Lactococcus lactis and Leuconostoc bacteriophages |
title_sort | high-throughput qpcr system for simultaneous quantitative detection of dairy lactococcus lactis and leuconostoc bacteriophages |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5365131/ https://www.ncbi.nlm.nih.gov/pubmed/28339484 http://dx.doi.org/10.1371/journal.pone.0174223 |
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