Real-Time PCR Assays for the Specific Detection of Field Balkan Strains of Lumpy Skin Disease Virus

Abstract

Lumpy skin di sease (LSD) is an important disease of cattle which is included in the OIE list of notifi able terrestrial animal diseases because of its great economic importance. The etiological agent is the Lumpy skin disease virus (LSDV).In the control of LSD attenuated strains of LSDV and SPPV are successfully used as vaccine strains in infected areas. In the case of vaccination policy, due to the possibility of mild or systemic post-vaccination reactions in vaccinated animals, the application of diagnostic procedures that will rapidly and specifi cally differentiate LSDV fi eld strains from LSD vaccine virus strains are extremely important. Rapidity in diagnostics and disposal of infected animals is one of the key factors in the prevention of spreading the disease.In the presented study we have described the development and validation of two realtime TaqMan-PCR assays for a rapid, sensitive and specifi c detection of the virulent fi eld LSDV strain currently circulating in the Balkan Peninsula. Specifi city for the fi eld strain and exclusivity for vaccine strains was tested on 171 samples from naturally infected and vaccinated animals.The results of this study show that both developed real-time PCR assays are more sensitive than the conventional nested PCR in detecting fi eld LSDV strains thus enabling rapid and high-throughput detection of animals infected with fi eld strains of LSDV.In conclusion, both KV-2 and FLI real-time PCR assays described in this study are simple, rapid, sensitive and suitable for routine use in a diagnostic laboratory and have the potential to replace conventional nested gel-based PCR assays as the standard procedure for the detection of fi eld strains of LSDV in clinical samples.

Epizootic features and control measures for lumpy skin disease in south‐east Serbia in 2016

“…At the beginning of the LSD epidemic in Serbia, a new assay for the detection of field strains of LSDV, KV‐2, was developed. This assay was introduced in the LSD laboratory confirmation procedure because it was easy to perform, very sensitive, specific and high‐throughput, so large numbers of samples could be tested during one day (Vidanovic et al, ).…”

“…All the samples from suspect animals were tested first for the presence of the LSDV genome using the capripox real‐time PCR assay according to Bowden, Babiuk, Parkyn, Coops, and Boyle (). The samples that were capripox positive were then tested with KV‐2 real‐time PCR assay, which is specific for the field strains of LSDV (Vidanovic et al, ). Animals which were positive in this assay were considered infected with a field strain of LSDV.…”

Total Number of Cattle in the Republic of Serbia after an Outbreak of Lumpy Skin Disease

“…Lumpy skin disease (LSD) is an important disease of cattle included in the OIE list of the notifiable terrestrial animal diseases because of its great economic importance (Vidanović, et al, 2016). During 2016 attention was particularly drawn to the situation in the Balkans and Eastern Europe, where there were cases of LSD and reports of the spread of the disease in Turkey, Greece, Bulgaria, Macedonia and Serbia.…”

“…Given that there is no specific treatment of LSD, appropriate preventive measures were implemented to control and prevent the further spread of the disease (VAMA, 2016a). The etiological agent of LSD is the lumpy skin disease virus (LSDV) (Vidanović, et al, 2016), which is not pathogenic to humans (Bowden et al, 2009;VAMA, 2016a).…”

“…As stipulated by some Regulations (OGRS, 2016), following reports of the first suspected case of infectious LSD in the municipality, the competent veterinary inspector, epizootiologist and veterinary authority are to perform clinical examination, conduct epizootic tests and inspect documents on the farm. Prompt disease diagnosis and safe disposal of infected animals are key factors in the prevention of the spread of the disease (Vidanović, et al, 2016).…”