Dea S, Michaud L, Milane G
Centre de recherche en virologie, Institut Armand-Frappier, Université du Québec, Laval, Canada.
J Gen Virol. 1995 May;76 ( Pt 5):1263-70. doi: 10.1099/0022-1317-76-5-1263.
Cytopathic coronaviruses were isolated in HRT-18 cells from bloody faecal samples collected from cows in Québec dairy herds having experienced typical outbreaks of winter dysentery (WD). The formation of polykaryons in the infected cell cultures was found to be dependent on the presence of trypsin in the medium. The WD isolates differed from the prototype Mebus strain of bovine enteropathogenic coronavirus (BCV.Meb) in respect to haemagglutination inhibition (HI), haemagglutination patterns at 4 degrees C and 37 degrees C, and receptor destroying enzyme activity with rat erythrocytes. Other field strains of BCV associated with outbreaks of neonatal calf diarrhoea (NCD) also differed from the BCV.Meb strain by demonstrating differences in HI. In all cases, no differences were detected by virus neutralization and Western immunoblotting. Analysis and comparison of the nucleotide and deduced amino acid sequences of the PCR-amplified haemagglutinin esterase (HE) genes of one representative WD strain (BCQ.2590) and two highly cytopathic NCD strains (BCQ.3 and BCQ.571) revealed high degrees of similarities (nt and aa sequence homologies > 98%) with the BCV.Meb strain. The putative esterase active site FGDS was conserved among these four BCV strains, indicating that this domain is probably not a determinant for BCV virulence. Six amino acid substitutions occurred between the HE glycoproteins of BCV.Meb and BCQ.2590 strains; two proline substitutions occurred respectively in the signal peptide (at aa 5) and near the sequences of the putative esterase domain (at aa 53).
从魁北克奶牛场经历典型冬季痢疾(WD)暴发的奶牛采集的血性粪便样本中,在HRT - 18细胞中分离出具有细胞病变效应的冠状病毒。发现在感染的细胞培养物中多核体的形成取决于培养基中胰蛋白酶的存在。WD分离株在血凝抑制(HI)、4℃和37℃时的血凝模式以及对大鼠红细胞的受体破坏酶活性方面与牛肠道致病性冠状病毒(BCV.Meb)的原型Mebus株不同。与新生犊牛腹泻(NCD)暴发相关的其他BCV野毒株在HI方面也与BCV.Meb株不同。在所有情况下,通过病毒中和试验和Western免疫印迹未检测到差异。对一株代表性WD毒株(BCQ.2590)和两株高细胞病变性NCD毒株(BCQ.3和BCQ.571)的PCR扩增血凝素酯酶(HE)基因的核苷酸和推导氨基酸序列进行分析和比较,发现与BCV.Meb株具有高度相似性(核苷酸和氨基酸序列同源性>98%)。这四株BCV毒株中推定的酯酶活性位点FGDS是保守的,表明该结构域可能不是BCV毒力的决定因素。BCV.Meb株和BCQ.2590株的HE糖蛋白之间发生了六个氨基酸替换;两个脯氨酸替换分别发生在信号肽(第5位氨基酸)和推定酯酶结构域序列附近(第53位氨基酸)。