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Open Access Research

Comparison of a loop-mediated isothermal amplification for orf virus with quantitative real-time PCR

Guangxiang Wang14, Youjun Shang34, Yanhua Wang14, Hong Tian2 and Xiangtao Liu14*

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 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

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Virology Journal 2013, 10:138  doi:10.1186/1743-422X-10-138

Published: 1 May 2013

Abstract

Background

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.

Methods

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.

Results

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%.

Conclusion

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.

Keywords:
Orf virus; Loop-mediated isothermal amplification (LAMP); Real-time PCR