Diagnostic Laboratory, School of Veterinary Medicine, Faculty of Health Sciences, Aristotle University of Thessaloniki, Thessaloniki, Greece; National Reference Laboratory for CaPVs, Department of Molecular Diagnostics, FMD, Virological, Rickettsial & Exotic Diseases, Athens Veterinary Center, Ministry of Rural Development and Food, Athens, Greece.
Diagnostic Laboratory, School of Veterinary Medicine, Faculty of Health Sciences, Aristotle University of Thessaloniki, Thessaloniki, Greece.
J Virol Methods. 2017 Nov;249:48-57. doi: 10.1016/j.jviromet.2017.08.011. Epub 2017 Aug 31.
Lumpy skin disease (LSD) is a transboundary viral disease of cattle with severe economic impact. Immunization of cattle with homologous live attenuated vaccines poses a number of diagnostic problems, as it has been associated with adverse reactions resembling disease symptoms. The latter hampers clinical diagnosis and poses challenges in virus identification. To this end, a duplex quantitative real-time PCR method targeting the GPCR gene was developed and validated, for the concurrent detection and differentiation of wild type and vaccine Lumpy skin disease virus (LSDV) strains. The method was evaluated in three laboratories. The evaluation included a panel of 38 poxvirus isolates/strains and the analytical characteristics of the method were determined. Amplification efficiencies were 91.3% and 90.7%, for wild type and vaccine LSDV, respectively; the limit of detection was 8 DNA copies for both targets and the inter-assay CV was 0.30% for wild type and 0.73% for vaccine LSDV. The diagnostic performance was assessed using 163 LSDV-positive samples, including field specimens and samples from experimentally vaccinated/infected animals. The method is able to confirm diagnosis in suspect cases, it differentiates infected from vaccinated animals (DIVA) and can be regarded as an important tool for effective LSD surveillance and eradication during vaccination campaigns.
块状皮肤病（LSD）是一种具有严重经济影响的牛传染性病毒病。用同源活减毒疫苗对牛进行免疫接种会带来一些诊断问题，因为它与类似于疾病症状的不良反应有关。后者妨碍了临床诊断，并在病毒鉴定方面带来了挑战。为此，开发并验证了一种针对 GPCR 基因的双定量实时 PCR 方法，用于同时检测和区分野生型和疫苗 LSD 病毒（LSDV）株。该方法在三个实验室进行了评估。该评估包括一组 38 种痘病毒分离株/株，确定了该方法的分析特性。野生型和疫苗 LSDV 的扩增效率分别为 91.3%和 90.7%；两种靶标检测限均为 8 个 DNA 拷贝，野生型和疫苗 LSDV 的组内变异系数分别为 0.30%和 0.73%。使用 163 份 LSDV 阳性样本评估了诊断性能，包括现场标本和来自实验接种/感染动物的样本。该方法能够在疑似病例中确认诊断，能够区分感染动物和接种疫苗动物（DIVA），并可作为在接种疫苗期间进行有效 LSD 监测和根除的重要工具。
