Comparison of a loop-mediated isothermal amplification for orf virus with quantitative real-time PCR
1 State Key Laboratory of Veterinary Etiological Biology, Lanzhou 730046, China
2 National Foot and Mouth Disease Reference Laboratory, Lanzhou 730046, China
3 Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou 730046, China
4 Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Science, Lanzhou 730046, China
Virology Journal 2013, 10:138 doi:10.1186/1743-422X-10-138Published: 1 May 2013
Orf virus (ORFV) causes orf (also known as contagious ecthyma or contagious papular dermatitis), a severe infectious skin disease in goats, sheep and other ruminants. Therefore, a rapid, highly specific and accurate method for the diagnosis of ORFV infections is essential to ensure that the appropriate treatments are administered and to reduce economic losses.
A loop-mediated isothermal amplification (LAMP) assay based on the identification of the F1L gene was developed for the specific detection of ORFV infections. The sensitivity and specificity of the LAMP assay were evaluated, and the effectiveness of this method was compared with that of real-time PCR.
The sensitivity of this assay was determined to be 10 copies of a standard plasmid. Furthermore, no cross-reactivity was found with either capripox virus or FMDV. The LAMP and real-time PCR assays were both able to detect intracutaneous- and cohabitation-infection samples, with a concordance of 97.83%. LAMP demonstrated a sensitivity of 89.13%.
The LAMP assay is a highly efficient and practical method for detecting ORFV infection. This LAMP method shows great potential for monitoring the prevalence of orf, and it could prove to be a powerful supplemental tool for current diagnostic methods.