Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia.
J Clin Microbiol. 2013 Nov;51(11):3818-25. doi: 10.1128/JCM.01531-13. Epub 2013 Sep 11.
Mammalian orthoreoviruses (MRVs) are known to cause mild enteric and respiratory infections in humans. They are widespread and infect a broad spectrum of mammals. We report here the first case of an MRV detected in a child with acute gastroenteritis, which showed the highest similarity to an MRV reported recently in European bats. An examination of a stool sample from the child was negative for most common viral and bacterial pathogens. Reovirus particles were identified by electron microscopic examination of both the stool suspension and cell culture supernatant. The whole-genome sequence was obtained with the Ion Torrent next-generation sequencing platform. Prior to sequencing, the stool sample suspension and cell culture supernatant were pretreated with nucleases and/or the convective interaction medium (CIM) monolithic chromatographic method to purify and concentrate the target viral nucleic acid. Whole-genome sequence analysis revealed that the Slovenian SI-MRV01 isolate was most similar to an MRV found in a bat in Germany. High similarity was shared in all genome segments, with nucleotide and amino acid identities between 93.8 to 99.0% and 98.4 to 99.7%, respectively. It was shown that CIM monolithic chromatography alone is an efficient method for enriching the sample in viral particles before nucleic acid isolation and next-generation sequencing application.
哺乳动物正呼肠孤病毒(MRV)已知可引起人类轻度肠和呼吸道感染。它们分布广泛,感染多种哺乳动物。我们在此报告首例在急性胃肠炎儿童中检测到的 MRV,该病毒与最近在欧洲蝙蝠中报道的 MRV 具有最高的相似性。对患儿粪便样本的检查排除了大多数常见的病毒和细菌病原体。通过电子显微镜检查粪便悬液和细胞培养上清液,鉴定出呼肠孤病毒颗粒。使用 Ion Torrent 下一代测序平台获得了全基因组序列。在测序之前,用核酸酶和/或对流相互作用介质(CIM)整体色谱法预处理粪便样本悬液和细胞培养上清液,以纯化和浓缩目标病毒核酸。全基因组序列分析表明,斯洛文尼亚 SI-MRV01 分离株与在德国蝙蝠中发现的 MRV 最为相似。所有基因组片段均具有高度相似性,核苷酸和氨基酸同一性分别为 93.8%至 99.0%和 98.4%至 99.7%。结果表明,在进行核酸分离和下一代测序应用之前,CIM 整体色谱法单独使用是一种有效富集样本中病毒颗粒的方法。