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
Descripción
Sumario: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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