Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Mei, Yunjun, He, Congcong, Deng, Wei, Ba, Dala, Yang, Ming, Zhang, Jian, Zhang, Shunxi, Shen, Ping, Chen, Xiangdong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
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
_version_ 1782432585173958656
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
work_keys_str_mv AT meiyunjun arealtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT hecongcong arealtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT dengwei arealtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT badala arealtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT yangming arealtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT zhangjian arealtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT zhangshunxi arealtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT shenping arealtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT chenxiangdong arealtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT meiyunjun realtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT hecongcong realtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT dengwei realtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT badala realtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT yangming realtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT zhangjian realtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT zhangshunxi realtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT shenping realtimepcrmethodtodetectthepopulationlevelofhalovirussnj1
AT chenxiangdong realtimepcrmethodtodetectthepopulationlevelofhalovirussnj1