Yazici Zafer, Ozan Emre, Tamer Cuneyt, Muftuoglu Bahadir, Barry Gerald, Kurucay Hanne Nur, Elhag Ahmed Eisa, Cagirgan Abdurrahman Anil, Gumusova Semra, Albayrak Harun
Department of Veterinary Virology, Faculty of Veterinary Medicine, Ondokuz Mayis University, 55139 Samsun, Turkey.
Department of Veterinary Experimental Animals, Faculty of Veterinary Medicine, Ondokuz Mayis University, 55139 Samsun, Turkey.
Animals (Basel). 2020 Sep 20;10(9):1700. doi: 10.3390/ani10091700.
Bovine respiratory disease (BRD) is a huge economic burden on the livestock industries of countries worldwide. Bovine respiratory syncytial virus (BRSV) is one of the most important pathogens that contributes to BRD. In this study, we report the identification and first isolation, with molecular characterization, of a new BRSV strain from lung specimens of three beef cows in Turkey that died from respiratory distress. After the screening of lung tissues for BRD-associated viruses using a multiscreen antigen-ELISA, a BRSV antigen was detected. This was then confirmed by real-time RT-PCR specific for BRSV. Following confirmation, virus isolation was conducted in MDBK cell cultures and clear CPE, including syncytia compatible with BRSV, were detected. RT-nested PCR, using F gene-specific primers, was performed on the cultured isolates, and the products were sequenced and deposited to Genbank with accession numbers MT179304, MT024766, and MT0244767. Phylogenetic analysis of these sequences indicated that the cattle were infected with BRSV from subgroup III and were closely related to previously identified American and Turkish strains, but contained some amino acid and nucleotide differences. This research paves the way for further studies on the molecular characteristics of natural BRSV isolates, including full genome analysis and disease pathogenesis, and also contributes to the development of robust national strategies against this virus.
牛呼吸道疾病(BRD)给全球各国的畜牧业带来了巨大的经济负担。牛呼吸道合胞病毒(BRSV)是导致BRD的最重要病原体之一。在本研究中,我们报告了从土耳其3头因呼吸窘迫死亡的肉牛的肺标本中鉴定并首次分离出一种新的BRSV毒株,并进行了分子特征分析。使用多筛抗原酶联免疫吸附测定法(ELISA)对肺组织进行BRD相关病毒筛查后,检测到了BRSV抗原。随后通过针对BRSV的实时逆转录聚合酶链反应(RT-PCR)进行了确认。确认后,在MDBK细胞培养物中进行病毒分离,检测到明显的细胞病变效应(CPE),包括与BRSV相符的多核巨细胞。使用F基因特异性引物对培养的分离株进行逆转录巢式PCR,对产物进行测序并提交至Genbank,登录号分别为MT179304、MT024766和MT0244767。对这些序列进行系统发育分析表明,这些牛感染了III亚群的BRSV,与先前鉴定的美国和土耳其毒株密切相关,但存在一些氨基酸和核苷酸差异。本研究为进一步研究天然BRSV分离株的分子特征(包括全基因组分析和疾病发病机制)铺平了道路,也有助于制定针对该病毒的有力国家战略。