Recombinase polymerase amplification assay for rapid detection of lumpy skin disease virus.
BACKGROUND
Lumpy 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.


RESULTS
The 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.


CONCLUSIONS
The 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.
Virology Department, Faculty of Veterinary Medicine, Cairo University, 12211, Giza, Egypt.
Mohamed A
MA
Shalaby
Virology Department, Faculty of Veterinary Medicine, Cairo University, 12211, Giza, Egypt.
Ayman
A
El-Deeb
Virology Department, Faculty of Veterinary Medicine, Mansoura University, 35516, Mansoura, Egypt.
Mohamed
M
El-Tholoth
Institute of Diagnostic Virology, Friedrich-Loeffler-Institute, 17493, Greifswald-Insel Riems, Germany.
Donata
D
Hoffmann
Division of Microbiology and Animal Hygiene, Department of Animal Sciences, Faculty of Agriculture Sciences, Georg-August-University, 37077, Goettingen, Germany.
Claus-Peter
CP
Czerny
Institute of Microbiology & Virology, Brandenburg Medical School Fontane, 01968, Senftenberg, Germany.
Frank T
FT
Hufert
Institute of Aquaculture, University of Stirling, FK9 4LA, Stirling, Scotland, UK.
Manfred
M
Weidmann
Division of Microbiology and Animal Hygiene, Department of Animal Sciences, Faculty of Agriculture Sciences, Georg-August-University, 37077, Goettingen, Germany. abdelwahed@gwdg.de.
Ahmed
A
Abd El Wahed
D000818:Animals; D002417:Cattle / Q000821:virology*; D004279:DNA, Viral; D008167:Lumpy skin disease virus / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity
D004279:DNA, Viral
Cattle; Lumpy skin disease virus; Point of need test; Recombinase polymerase amplification assay