Development of a SYBR green real-time PCR method for rapid detection of sheep pox virus
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
Virology Journal 2012, 9:291 doi:10.1186/1743-422X-9-291Published: 27 November 2012
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.
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.
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.