Menasherow Sophia, Erster Oran, Rubinstein-Giuni Marisol, Kovtunenko Anita, Eyngor Evgeny, Gelman Boris, Khinich Evgeny, Stram Yehuda
Virology Division, Kimron Veterinary Institute, P.O. Box 12, Bet Dagan 50250, Israel.
Virology Division, Kimron Veterinary Institute, P.O. Box 12, Bet Dagan 50250, Israel.
J Virol Methods. 2016 Jun;232:12-5. doi: 10.1016/j.jviromet.2016.02.008. Epub 2016 Feb 20.
Lumpy skin disease (LSD) is a constant threat to the Middle East including the State of Israel. During vaccination programs it is essential for veterinary services and farmers to be able to distinguish between animals affected by the cattle-borne virulent viruses and vaccinated animals, subsequently affected by the vaccine strain. This study describes an improved high resolution-melting (HRM) test that exploits a 27 base pair (bp) fragment of the LSDV126 extracellular enveloped virion (EEV) gene that is present in field viruses but is absent from the Neethling vaccine strain. This difference leads to ∼0.5 °C melting point change in the HRM assay, when testing the quantitative PCR (qPCR) products generated from the virulent field viruses compared to the attenuated vaccine. By exploiting this difference, it could be shown using the newly developed HRM assay that virus isolated from vaccinated cattle that developed disease symptoms behave similarly to vaccine virus control, indicating that the vaccine virus can induce disease symptoms. This assay is not only in full agreement with the previously published PCR gradient and restriction fragment length polymorphism (RFLP) tests but it is faster with, fewer steps, cheaper and dependable.
结节性皮肤病（LSD）对包括以色列国在内的中东地区构成持续威胁。在疫苗接种计划期间，兽医服务部门和养殖户能够区分受牛传播的强毒病毒感染的动物和随后受疫苗毒株影响的接种疫苗动物至关重要。本研究描述了一种改进的高分辨率熔解（HRM）检测方法，该方法利用了LSDV126细胞外包膜病毒粒子（EEV）基因的一个27碱基对（bp）片段，该片段存在于野外病毒中，但在Neethling疫苗毒株中不存在。当检测从强毒野外病毒与减毒疫苗产生的定量PCR（qPCR）产物时，这种差异导致HRM分析中的熔点变化约0.5℃。通过利用这种差异，使用新开发的HRM检测方法可以表明，从出现疾病症状的接种疫苗牛中分离出的病毒与疫苗病毒对照表现相似，这表明疫苗病毒可以诱发疾病症状。该检测方法不仅与先前发表的PCR梯度和限制性片段长度多态性（RFLP）检测完全一致，而且速度更快、步骤更少、成本更低且可靠。
