Department of Electron Microscopy and Histopathology, Noguchi Memorial Institute for Medical Research, College of Health Sciences, University of Ghana, Legon P.O. Box LG581, Ghana.
Department of Hygiene and Molecular Epidemiology, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan.
Viruses. 2024 Sep 27;16(10):1529. doi: 10.3390/v16101529.
Most human rotaviruses belong to the Wa-like, DS-1-like, or AU-1-like genotype constellation. The AU-1-like constellation, albeit minor, captured attention because its prototype strain AU-1 originated from feline rotavirus, leading to the concept of interspecies transmission of rotavirus. The AU-1 genome sequence determined by various laboratories over the years has documented two conflicting VP7 sequences in the GenBank. As culture-adaptation may introduce changes in the viral genome, the original fecal (wild-type) and the seed stock of culture-adapted AU-1 genomes were sequenced using the Illumina's MiSeq platform to determine the authentic AU-1 sequence and to identify what mutational changes were selected during cell-culture adaptation. The wild-type and culture-adapted AU-1 genomes were identical except for one VP4-P475L substitution. Their VP7 gene was 99.9% identical to the previously reported AU-1 VP7 under accession number AB792641 but only 92.5% to that under accession number D86271. Thus, the wild-type sequences determined in this study (accession numbers OR727616-OR727626) should be used as the reference. The VP4-P475L mutation was more likely incidental than inevitable during cell-culture adaptation. This was the first study in which the whole genomes of both wild-type and cultured RVA strains were simultaneously determined by deep sequencing.
大多数人类轮状病毒属于 Wa 样、DS-1 样或 AU-1 样基因型组合。AU-1 样组合虽然很小,但引起了人们的关注,因为其原型株 AU-1 来源于猫轮状病毒,导致了轮状病毒的种间传播的概念。多年来,不同实验室确定的 AU-1 基因组序列在 GenBank 中记录了两个相互冲突的 VP7 序列。由于培养适应可能会导致病毒基因组发生变化,因此使用 Illumina 的 MiSeq 平台对原始粪便(野生型)和适应培养的 AU-1 基因组的种子库进行测序,以确定真实的 AU-1 序列,并确定在细胞培养适应过程中选择了哪些突变变化。野生型和适应培养的 AU-1 基因组除了一个 VP4-P475L 取代外完全相同。它们的 VP7 基因与先前报道的登录号 AB792641 下的 AU-1 VP7 同源性为 99.9%,但与登录号 D86271 下的同源性仅为 92.5%。因此,本研究中确定的野生型序列(登录号 OR727616-OR727626)应作为参考。VP4-P475L 突变在细胞培养适应过程中更可能是偶然发生的,而不是必然发生的。这是第一项通过深度测序同时确定野生型和培养的 RVA 株全基因组的研究。