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BMC Veterinary Research

, 12:244

Virology

Abstract

BackgroundLumpy skin disease virus LSDV is a Capripoxvirus infecting cattle and Buffalos. Lumpy skin disease LSD leads to significant economic losses due to hide damage, reduction of milk production, mastitis, infertility and mortalities 10 %. Early detection of the virus is crucial to start appropriate outbreak control measures. Veterinarians rely on the presence of the characteristic clinical signs of LSD. Laboratory diagnostics including virus isolation, sequencing and real-time polymerase chain reaction PCR are performed at well-equipped laboratories. In this study, a portable, simple, and rapid recombinase polymerase amplification RPA assay for the detection of LSDV-genome for the use on farms was developed.

ResultsThe LSDV RPA assay was performed at 42 °C and detected down to 179 DNA copies-reaction in a maximum of 15 min. Unspecific amplification was observed with neither LSDV-negative samples n = 12 nor nucleic acid preparations from orf virus, bovine papular stomatitis virus, cowpoxvirus, Peste des petits ruminants and Blue tongue virus serotypes 1, 6 and 8. The clinical sensitivity of the LSDV RPA assay matched 100 % n = 22 to real-time PCR results. In addition, the LSDV RPA assay detected sheep and goat poxviruses.

ConclusionThe LSDV RPA assay is a rapid and sensitive test that could be implemented in field or at quarantine stations for the identification of LSDV infected case.

KeywordsLumpy skin disease virus Recombinase polymerase amplification assay Point of need test Cattle AbbreviationsFPForward primer

GPCRG-protein-coupled chemokine receptor

GPVGoat poxvirus

LAMPLoop-mediated isothermal amplification

LSDLumpy skin disease

LSDVLumpy skin disease virus

nNumber

OIEOffice International des Epizooties

PCRPolymerase chain reaction

RPReverse primer

RPARecombinase polymerase amplification

SPVSheep poxvirus

Electronic supplementary materialThe online version of this article doi:10.1186-s12917-016-0875-5 contains supplementary material, which is available to authorized users.

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Autor: Mohamed A. Shalaby - Ayman El-Deeb - Mohamed El-Tholoth - Donata Hoffmann - Claus-Peter Czerny - Frank T. Hufert - Manfred

Fuente: https://link.springer.com/







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