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Development of a SYBR green real-time PCR method for rapid detection of sheep pox virus
Hong Tian1, Jingyan Wu, Yan Chen
1State Key Laboratory of Veterinary Etiological Biology, Lanzhou 730046, China.
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.
