Abstract
BackgroundOrf virus (ORFV) is the causative agent of a severe infectious skin disease (also known as contagious ecthyma) in goats, sheep and other small ruminants. Importantly, ORFV also infect humans which causes a public health concern in the context of changing environment and increase in human populations. The rapid detection is critical in effective control of the disease and urgently needed.ResultsA novel “point of care” molecular amplification assay for rapid visual detection of ORFV was developed based on isothermoal recombinase polymerase amplification (RPA) technology in combination with a simpler lateral flow immunoassay strip (ORFV RPA- LFD assay). The developed ORFV RPA- LFD assay was able to detect ORFV in less than 25 min. This assay was highly sensitive, with detection limit of as low as 80 copies/reaction, and highly specific, with no cross-reactions with capripox virus, foot-and-mouth disease virus and peste des petits ruminants virus. Furthermore, the ORFV RPA- LFD assay has good correlation with qPCR assay for detection of ORFV present in clinical samples.ConclusionsThe developed ORFV RPA-LFD assay was a sensitive and specific method for rapid detection of ORFV, and has great potential as an onsite molecular diagnostic tool in control of Orf.Electronic supplementary materialThe online version of this article (doi:10.1186/s12985-016-0502-x) contains supplementary material, which is available to authorized users.
Highlights
Orf virus (ORFV) is the causative agent of a severe infectious skin disease in goats, sheep and other small ruminants
Orf, known as contagious ecthyma, is a skin disease caused by Orf virus (ORFV), which is classified in the genus Parapoxvirus of the family Poxviridae
Optimization of the ORFV recombinase polymerase amplification (RPA)-lateral flow dipstick (LFD) conditions To determine the optimal temperature for the RPA reaction, the ability of the ORFV RPA–LFD assay was tested to amplify 80 copies of ORFV DNA as template at a range of temperatures from 15 °C to 50 °C
Summary
Orf virus (ORFV) is the causative agent of a severe infectious skin disease ( known as contagious ecthyma) in goats, sheep and other small ruminants. ORFV infect humans which causes a public health concern in the context of changing environment and increase in human populations. Known as contagious ecthyma, is a skin disease caused by Orf virus (ORFV), which is classified in the genus Parapoxvirus of the family Poxviridae. Humans of infection with ORFV is thought to be under diagnosed and causes a public health concern in the context of. Molecular amplification assays, in particular real-time quantitative PCR (qPCR) and loopmediated isothermal amplification (LAMP) are sensitive for detection of ORFV [2,3,4,5]. A novel and rapid isothermal molecular diagnostic approach, termed recombinase polymerase amplification(RPA), has been developed as an alternative to PCR assay and
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