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Open Access Highly Accessed Short report

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

Hong Tian1, Jingyan Wu2, Yan Chen3, Keshan Zhang4, Youjun Shang4 and Xiangtao Liu3*

Author Affiliations

1 State Key Laboratory of Veterinary Etiological Biology, Lanzhou, 730046, China

2 National Foot and Mouth Disease Reference Laboratory, Lanzhou, 730046, China

3 Lanzhou Veterinary Research Institute, Yanchangbu, Xujiaping, Lanzhou, 730046, China

4 Chinese Academy of Agricultural Sciences, Lanzhou, 730046, China

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Virology Journal 2012, 9:291  doi:10.1186/1743-422X-9-291

Published: 27 November 2012

Abstract

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 101 to 109 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.

Keywords:
Sheep pox virus; SYBR Green I based quantitative PCR