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A Real-Time PCR Method to Detect the Population Level of Halovirus SNJ1
Although viruses of haloarchaea are the predominant predator in hypersaline ecosystem, the culture studies about halovirus-host systems are infancy. The main reason is the tradition methodology (plaque assay) for virus-host interaction depends on culturable and susceptible host. Actually, more than...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4871410/ https://www.ncbi.nlm.nih.gov/pubmed/27192212 http://dx.doi.org/10.1371/journal.pone.0155642 |
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author | Mei, Yunjun He, Congcong Deng, Wei Ba, Dala Yang, Ming Zhang, Jian Zhang, Shunxi Shen, Ping Chen, Xiangdong |
author_facet | Mei, Yunjun He, Congcong Deng, Wei Ba, Dala Yang, Ming Zhang, Jian Zhang, Shunxi Shen, Ping Chen, Xiangdong |
author_sort | Mei, Yunjun |
collection | PubMed |
description | Although viruses of haloarchaea are the predominant predator in hypersaline ecosystem, the culture studies about halovirus-host systems are infancy. The main reason is the tradition methodology (plaque assay) for virus-host interaction depends on culturable and susceptible host. Actually, more than 90% of haloarchaea are unculturable. Therefore, it is necessary to establish an approach for detecting the dynamics of virus in hypersaline environment without culture. In this study, we report a convenient method to determine the dynamics of halovirus SNJ1 based on quantitative real-time PCR (qPCR). All findings showed that the qPCR method was specific (single peak in melt curves), accurate (a good linear relationship between the log of the PFU and the C(t) values, R(2) = 0.99), reproducible (low coefficient of variations, below 1%). Additionally, the physicochemical characteristics of the samples tested did not influence the stability of qPCR. Therefore, the qPCR method has the potential value in quantifying and surveying haloviruses in halophilic ecological system. |
format | Online Article Text |
id | pubmed-4871410 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-48714102016-05-31 A Real-Time PCR Method to Detect the Population Level of Halovirus SNJ1 Mei, Yunjun He, Congcong Deng, Wei Ba, Dala Yang, Ming Zhang, Jian Zhang, Shunxi Shen, Ping Chen, Xiangdong PLoS One Research Article Although viruses of haloarchaea are the predominant predator in hypersaline ecosystem, the culture studies about halovirus-host systems are infancy. The main reason is the tradition methodology (plaque assay) for virus-host interaction depends on culturable and susceptible host. Actually, more than 90% of haloarchaea are unculturable. Therefore, it is necessary to establish an approach for detecting the dynamics of virus in hypersaline environment without culture. In this study, we report a convenient method to determine the dynamics of halovirus SNJ1 based on quantitative real-time PCR (qPCR). All findings showed that the qPCR method was specific (single peak in melt curves), accurate (a good linear relationship between the log of the PFU and the C(t) values, R(2) = 0.99), reproducible (low coefficient of variations, below 1%). Additionally, the physicochemical characteristics of the samples tested did not influence the stability of qPCR. Therefore, the qPCR method has the potential value in quantifying and surveying haloviruses in halophilic ecological system. Public Library of Science 2016-05-18 /pmc/articles/PMC4871410/ /pubmed/27192212 http://dx.doi.org/10.1371/journal.pone.0155642 Text en © 2016 Mei 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 Mei, Yunjun He, Congcong Deng, Wei Ba, Dala Yang, Ming Zhang, Jian Zhang, Shunxi Shen, Ping Chen, Xiangdong A Real-Time PCR Method to Detect the Population Level of Halovirus SNJ1 |
title | A Real-Time PCR Method to Detect the Population Level of Halovirus SNJ1 |
title_full | A Real-Time PCR Method to Detect the Population Level of Halovirus SNJ1 |
title_fullStr | A Real-Time PCR Method to Detect the Population Level of Halovirus SNJ1 |
title_full_unstemmed | A Real-Time PCR Method to Detect the Population Level of Halovirus SNJ1 |
title_short | A Real-Time PCR Method to Detect the Population Level of Halovirus SNJ1 |
title_sort | real-time pcr method to detect the population level of halovirus snj1 |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4871410/ https://www.ncbi.nlm.nih.gov/pubmed/27192212 http://dx.doi.org/10.1371/journal.pone.0155642 |
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