Oliver S L, Batten C A, Deng Y, Elschner M, Otto P, Charpilienne A, Clarke I N, Bridger J C, Lambden P R
Department of Pathology and Infectious Diseases, Royal Veterinary College, Camden, London, United Kingdom.
J Clin Microbiol. 2006 Mar;44(3):992-8. doi: 10.1128/JCM.44.3.992-998.2006.
The bovine enteric caliciviruses Bo/Jena/1980/DE and Bo/Newbury2/1976/UK represent two distinct genotypes within a new genogroup, genogroup III, in the genus Norovirus of the family Caliciviridae. In the present study, the antigenic relatedness of these two genotypes was determined for the first time to enable the development of tests to detect and differentiate between both genotypes. Two approaches were used. First, cross-reactivity was examined by enzyme-linked immunosorbent assay (ELISA) using recombinant virus-like particles (VLPs) and convalescent-phase sera from calves infected with either Jena (genotype 1) or Newbury2 (genotype 2). Second, cross-reactivity was examined between the two genotypes with a monoclonal antibody, CM39, derived using Jena VLPs. The two genotypes, Jena and Newbury2, were antigenically distinct with little or no cross-reactivity by ELISA to the heterologous VLPs using convalescent calf sera that had homologous immunoglobulin G titers of log10 3.1 to 3.3. CM39 reacted with both Jena and heterologous Newbury2 VLPs. The CM39 epitope was mapped to nine amino acids (31PTAGAQIAA39) in the Jena capsid protein, which was not fully conserved for Newbury2 (31PTAGAPVAA39). Molecular modeling showed that the CM39 epitope was located within the NH2-terminal arm inside the virus capsid. Surprisingly, CM39 also reacted with VLPs from two genogroup II/3 human noroviruses by ELISA and Western blotting. Thus, although the bovine noroviruses Jena and Newbury2 corresponded to two distinct antigenic types or serotypes, they shared at least one cross-reactive epitope. These findings have relevance for epidemiological studies to determine the prevalence of bovine norovirus serotypes and to develop vaccines to bovine noroviruses.
牛肠道杯状病毒Bo/Jena/1980/DE和Bo/Newbury2/1976/UK代表杯状病毒科诺如病毒属新基因组(基因组III)内的两个不同基因型。在本研究中,首次确定了这两种基因型的抗原相关性,以便开发检测和区分这两种基因型的检测方法。采用了两种方法。首先,使用重组病毒样颗粒(VLP)和来自感染耶拿株(基因型1)或纽伯里2株(基因型2)的犊牛的恢复期血清,通过酶联免疫吸附测定(ELISA)检测交叉反应性。其次,使用源自耶拿株VLP的单克隆抗体CM39检测两种基因型之间的交叉反应性。耶拿株和纽伯里2株这两种基因型在抗原性上是不同的,使用同源免疫球蛋白G滴度为log10 3.1至3.3的犊牛恢复期血清通过ELISA检测,对异源VLP几乎没有或没有交叉反应性。CM39与耶拿株和异源纽伯里2株VLP均发生反应。CM39表位被定位到耶拿株衣壳蛋白中的九个氨基酸(31PTAGAQIAA39),纽伯里2株(31PTAGAPVAA39)对此并不完全保守。分子建模表明,CM39表位位于病毒衣壳内的NH2末端臂内。令人惊讶的是,通过ELISA和蛋白质印迹法检测,CM39也与两种基因组II/3型人诺如病毒的VLP发生反应。因此,尽管牛诺如病毒耶拿株和纽伯里2株对应于两种不同的抗原类型或血清型,但它们至少共享一个交叉反应表位。这些发现对于确定牛诺如病毒血清型的流行率以及开发牛诺如病毒疫苗的流行病学研究具有重要意义。