Animal Production and Health Laboratory, Joint FAO/IAEA Agricultural and Biotechnology laboratory, Division of Nuclear Techniques in Food and Agriculture, Department of Nuclear Sciences and Applications, International Atomic Energy Agency, Wagramer Strasse 5, P.O. Box 100, A1400, Vienna, Austria.
Institute of Biotechnology, University of Natural Resources and Life Sciences (BOKU), Muthgasse 18, 1190, Vienna, Austria.
Sci Rep. 2019 Apr 30;9(1):6646. doi: 10.1038/s41598-019-43158-x.
Sheep poxvirus (SPPV), goat poxvirus (GTPV) and lumpy skin disease virus (LSDV) affect small ruminants and cattle causing sheeppox (SPP), goatpox (GTP) and lumpy skin disease (LSD) respectively. In endemic areas, vaccination with live attenuated vaccines derived from SPPV, GTPV or LSDV provides protection from SPP and GTP. As live poxviruses may cause adverse reactions in vaccinated animals, it is imperative to develop new diagnostic tools for the differentiation of SPPV field strains from attenuated vaccine strains. Within the capripoxvirus (CaPV) homolog of the variola virus B22R gene, we identified a unique region in SPPV vaccines with two deletions of 21 and 27 nucleotides and developed a High-Resolution Melting (HRM)-based assay. The HRM assay produces four distinct melting peaks, enabling the differentiation between SPPV vaccines, SPPV field isolates, GTPV and LSDV. This HRM assay is sensitive, specific, and provides a cost-effective means for the detection and classification of CaPVs and the differentiation of SPPV vaccines from SPPV field isolates.
绵羊痘病毒（SPPV）、山羊痘病毒（GTPV）和牛结节疹病毒（LSDV）分别感染小反刍动物和牛，导致绵羊痘（SPP）、山羊痘（GTP）和牛结节疹病（LSD）。在流行地区，使用源自 SPPV、GTPV 或 LSDV 的弱毒活疫苗对小反刍动物和牛进行免疫接种，可预防 SPP 和 GTP。由于活痘病毒可能在接种动物中引起不良反应，因此开发新的诊断工具来区分 SPPV 田间株与弱毒疫苗株势在必行。在天花病毒 B22R 基因的羊痘病毒（CaPV）同源物中，我们在 SPPV 疫苗中发现了一个独特的区域，该区域有 21 和 27 个核苷酸的缺失，并开发了一种基于高分辨率熔解（HRM）的检测方法。HRM 检测法产生四个独特的熔解峰，可区分 SPPV 疫苗、SPPV 田间分离株、GTPV 和 LSDV。该 HRM 检测法具有敏感性、特异性，并且为检测和分类 CaPV 以及区分 SPPV 疫苗和 SPPV 田间分离株提供了一种具有成本效益的方法。
