Development of a SYBR green real-time PCR method for rapid detection of sheep pox virus

BACKGROUND: In this study, we developed a SYBR Green-based real-time PCR assay for the detection of sheep pox virus using a plasmid construct carrying one of the highly conserved genes encoding the virion envelope protein (P32) as a template. RESULTS: The method was demonstrated to be highly sensiti...

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Detalles Bibliográficos
Autores principales: Tian, Hong, Wu, Jingyan, Chen, Yan, Zhang, Keshan, Shang, Youjun, Liu, Xiangtao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Short Report
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3546957/
https://www.ncbi.nlm.nih.gov/pubmed/23186407
http://dx.doi.org/10.1186/1743-422X-9-291
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author Tian, Hong
Wu, Jingyan
Chen, Yan
Zhang, Keshan
Shang, Youjun
Liu, Xiangtao
author_facet Tian, Hong
Wu, Jingyan
Chen, Yan
Zhang, Keshan
Shang, Youjun
Liu, Xiangtao
author_sort Tian, Hong
collection PubMed
description BACKGROUND: In this study, we developed a SYBR Green-based real-time PCR assay for the detection of sheep pox virus using a plasmid construct carrying one of the highly conserved genes encoding the virion envelope protein (P32) as a template. RESULTS: The method was demonstrated to be highly sensitive, allowing a precise SPV DNA quantitation over a range of nine orders of magnitude (from 10(1) to 10(9) copies of standard DNA). Then, specimens from SPV suspected sheep were analyzed by conventional gel-based PCR, real-time PCR and sequence analysis. CONCLUSION: Comparison between these different techniques revealed that real-time PCR is more sensitive than conventional gel-based PCR, allowing detection low viral titers of SPV in infected sheep.
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spelling pubmed-35469572013-01-17 Development of a SYBR green real-time PCR method for rapid detection of sheep pox virus Tian, Hong Wu, Jingyan Chen, Yan Zhang, Keshan Shang, Youjun Liu, Xiangtao Virol J Short Report BACKGROUND: In this study, we developed a SYBR Green-based real-time PCR assay for the detection of sheep pox virus using a plasmid construct carrying one of the highly conserved genes encoding the virion envelope protein (P32) as a template. RESULTS: The method was demonstrated to be highly sensitive, allowing a precise SPV DNA quantitation over a range of nine orders of magnitude (from 10(1) to 10(9) copies of standard DNA). Then, specimens from SPV suspected sheep were analyzed by conventional gel-based PCR, real-time PCR and sequence analysis. CONCLUSION: Comparison between these different techniques revealed that real-time PCR is more sensitive than conventional gel-based PCR, allowing detection low viral titers of SPV in infected sheep. BioMed Central 2012-11-27 /pmc/articles/PMC3546957/ /pubmed/23186407 http://dx.doi.org/10.1186/1743-422X-9-291 Text en Copyright ©2012 Tian et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Short Report
Tian, Hong
Wu, Jingyan
Chen, Yan
Zhang, Keshan
Shang, Youjun
Liu, Xiangtao
Development of a SYBR green real-time PCR method for rapid detection of sheep pox virus
title Development of a SYBR green real-time PCR method for rapid detection of sheep pox virus
title_full Development of a SYBR green real-time PCR method for rapid detection of sheep pox virus
title_fullStr Development of a SYBR green real-time PCR method for rapid detection of sheep pox virus
title_full_unstemmed Development of a SYBR green real-time PCR method for rapid detection of sheep pox virus
title_short Development of a SYBR green real-time PCR method for rapid detection of sheep pox virus
title_sort development of a sybr green real-time pcr method for rapid detection of sheep pox virus
topic Short Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3546957/
https://www.ncbi.nlm.nih.gov/pubmed/23186407
http://dx.doi.org/10.1186/1743-422X-9-291
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