Molecular and antigen detection, phylogenetics, and immunoinformatics study of the zoonotic coronavirus in Iranian diarrheic calves

MohammadM Ranjbar, Soodeh Enayati, Soodeh Hooshmandi, Shahrzad Ahangarzadeh, Shima Aboutalebian
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Abstract

Background: Bovine coronaviruses (BCoVs) are zoonotic diseases that result in substantial economic losses due to mortality, impaired growth, and increased medication expenses in large animals. These viruses pose a risk to children who live beside infected animal, as they can cause diarrhea. This study was dedicated to molecular and antigen detections and phylogenetic and immunoinformatics analysis of zoonotic coronavirus (CoV) in Iran.Materials and Methods: A total of 77 diarrheic samples were collected from Holstein dairy herds in selected provinces of Iran. Samples were tested by capture antibody enzyme-linked immunosorbent assay (ELISA) to detect CoV and reverse transcriptase–polymerase chain reaction (RT–PCR) for verification of detection and also genotyping of spike glycoprotein in CoV-positive samples. After statistical analysis, nucleotide sequence alignment, and nucleotide and protein phylogenetic tree construction, the centralized sequence for vaccine strains was obtained using computationally optimized broadly reactive antigen (COBRA)'s center-of-the-tree (COT) method.Results: Twenty-two (28.5%) and eight (10.3%) of 77 samples were positive according to RT–PCR and ELISA, respectively. (Basic Local Alignment Search Tool) BLAST and phylogenetic analysis revealed that most similar sequences to the Iranian CoV sequence were for European countries. Furthermore, there were strong correlations to other CoVs in humans and wild and domesticated animals. As CoV has variable COT, the most recent strains and COBRA vaccine strains were obtained.Conclusion: Based on the high prevalence of this viral disease in calves and its economic impact on the breeding industry, as well as the potential transmission to humans and correlation with World Health Organization (WHO) One Health approach guidelines, the study emphasizes the importance of implementing preventive strategies such as animal vaccination.
伊朗腹泻小牛人畜共患冠状病毒的分子和抗原检测、系统发育和免疫信息学研究
背景:牛冠状病毒(BCoVs)是一种人畜共患疾病,由于大型动物的死亡率、生长受损和药物费用增加而导致重大经济损失。这些病毒对与受感染动物生活在一起的儿童构成风险,因为它们可能导致腹泻。本研究致力于伊朗人畜共患冠状病毒(CoV)的分子和抗原检测以及系统发育和免疫信息学分析。材料与方法:从伊朗选定省份的荷斯坦奶牛群中采集77例腹泻样本。用捕获抗体酶联免疫吸附试验(ELISA)检测冠状病毒,用逆转录聚合酶链反应(RT-PCR)验证冠状病毒阳性样本刺突糖蛋白的检测结果和基因分型。经统计分析、核苷酸序列比对、构建核苷酸和蛋白质系统发育树后,采用计算优化的广义反应性抗原(COBRA)树中心法(COT)获得疫苗株集中序列。结果:77份样品中,RT-PCR阳性22例(28.5%),ELISA阳性8例(10.3%)。BLAST和系统发育分析显示,与伊朗冠状病毒序列最相似的序列来自欧洲国家。此外,与人类以及野生和驯养动物中的其他冠状病毒有很强的相关性。由于冠状病毒具有可变的COT,因此获得了最新的株和COBRA疫苗株。结论:鉴于这种病毒性疾病在犊牛中的高流行率及其对养殖业的经济影响、对人类的潜在传播以及与世界卫生组织(WHO)“同一个健康”方针的相关性,本研究强调了实施动物疫苗接种等预防策略的重要性。
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