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Abstract：
Lumpy skin disease virus (LSDV) infection, accompanied by loss of hide quality, poor reproductive efficiency, consistent degenerative emaciation, and milk yield reduction of animals, causes severe economic implications in endemic zones. The heterologous attenuated goat pox (GTPV) vaccine (AV41 strain) was used in China to prevent LSDV infection. Only a few LSDV detection methods that distinguish LSDV from GTPV vaccine strains have been reported before. For simple, rapid, and specific detection of LSDV, the real-time recombinase polymerase amplification (RPA) method was established with the specific primers and probes designed according to the conserved regions of ORF132 gene sequences. The assay could be finished within 20 min at a constant temperature (39 °C). This method had a limit of detection (LOD) of 15 copies/μL for LSDV and no cross-reaction with the nucleic acids of goat pox virus, infectious bovine rhinotracheitis virus, Pasteurella multocida, and bovine healthy tissue. Furthermore, 43 clinical samples were detected by this method and the real-time PCR recommended by the World Organisation for Animal Health (WOAH), with a kappa value, was 0.94. These results demonstrated that the real-time RPA method for detecting LSDV developed in this study was characterized by high sensitivity and specificity, which has wide application value in the clinical diagnosis and detection of LSDV in China.
摘要：
结节性皮肤病病毒（LSDV）感染，伴随着皮革质量的损失，繁殖效率低，一致的退行性消瘦，和减少动物的产奶量，对流行区造成严重的经济影响。在中国使用异源减毒山羊痘（GTPV）疫苗（AV41株）来预防LSDV感染。以前仅报道了将LSDV与GTPV疫苗株区分开的几种LSDV检测方法。对于简单的，快速,和LSDV的特异性检测，根据ORF132基因序列的保守区设计特异性引物和探针，建立了实时重组酶聚合酶扩增（RPA）方法。测定可以在恒温(39°C)下在20分钟内完成。该方法对LSDV的检测限（LOD）为15个拷贝/μL，并且与山羊痘病毒的核酸没有交叉反应。传染性牛鼻支气管炎病毒，多杀性巴氏杆菌,和牛健康组织。此外，通过该方法和世界动物卫生组织（WOAH）推荐的实时PCR检测了43个临床样品，具有卡帕值，是0.94。这些结果表明，本研究开发的用于检测LSDV的实时RPA方法具有高敏感性和特异性，在我国LSDV的临床诊断和检测中具有广泛的应用价值。
