Rapid Molecular Detection of Genetically Diverted Foot and Mouth Disease Virus Serotype O During the Outbreak of 2012 in Egypt

S. Rahman, B. Hoffmann, B. Haas, Mohammed El Beskawy, M. Othman, M. Eladl
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Abstract

Increasing the international trade of animals and their products and the continuous changing of Foot and Mouth Disease Virus (FMDV) lead to the rise of disease introductions and create an urgent need for laboratory methods facilitating a swift and sensitive confirmation of suspect outbreaks and fast characterization of new isolates. As FMDV is extremely contagious and affects all cloven-hoofed animals, it provides a continuous burden and risk to the livestock industries of the developing world, in particular due to mortality in young animals, weight and milk loss, lameness and the trade restrictions necessary to control the disease. The control of FMD in Egypt requires an accurate analysis of the newly introduced viral strains and an analysis of their relationship with the current circulating strains and the routinely used vaccines. This study evaluates a recently developed rapid Reverse-Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) for the identification of FMDV in samples from suspect cases. Samples were collected from gum, tongue epithelium, vesicular fluid, buccal and gum swabs, as well as saliva of infected cattle, buffalo and sheep and examined using two RT-qPCR of routine and high-speed formats using "IRES1" and "3D-OIE" primers. In addition, partial VP1 sequencing of some selected isolates was carried out for phylogenetic analysis. The high-speed RT-qPCR allowed a reliable diagnosis of FMDV in less than half an hour and can be used as a fast and valuable method for the monitoring and controlling of the foot and mouth disease. A new variant genotype (FMD-O/Eg/Mans/14) was circulating in Egypt during the outbreak of 2012 showing 98.5% identity to the isolated strain in Sudan (SUD_6_2008).
2012年埃及爆发口蹄疫O型基因转移病毒的快速分子检测
动物及其产品国际贸易的增加以及口蹄疫病毒(FMDV)的不断变化导致疾病引进的增加,并迫切需要实验室方法,以便迅速和敏感地确认疑似疫情并快速表征新的分离株。由于口蹄疫病毒极具传染性,并影响所有偶蹄动物,它给发展中世界的畜牧业带来持续的负担和风险,特别是由于幼畜死亡、体重和乳汁损失、跛行以及为控制该疾病所必需的贸易限制。在埃及控制口蹄疫需要对新传入的病毒株进行准确分析,并分析它们与当前流行的病毒株和常规使用的疫苗之间的关系。本研究评估了最近开发的快速反转录定量聚合酶链反应(RT-qPCR),用于鉴定疑似病例样本中的口蹄疫病毒。从感染牛、水牛和羊的牙龈、舌上皮、囊泡液、口腔和牙龈拭子以及唾液中采集样本,使用“IRES1”和“3D-OIE”引物进行常规和高速两种RT-qPCR检测。此外,对部分分离株进行VP1部分测序,进行系统发育分析。高速RT-qPCR可在不到半小时的时间内对口蹄疫进行可靠诊断,可作为监测和控制口蹄疫的一种快速而有价值的方法。2012年暴发期间,一种新的变异基因型(FMD-O/Eg/Mans/14)在埃及流行,与苏丹分离的菌株(SUD_6_2008)有98.5%的同源性。
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