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J Clin Microbiol ; 51(11): 3818-25, 2013 Nov.
Article in English | MEDLINE | ID: mdl-24025904

ABSTRACT

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.


Subject(s)
Gastroenteritis/virology , Orthoreovirus/classification , Orthoreovirus/genetics , Reoviridae Infections/virology , Animals , Chiroptera/virology , Cluster Analysis , Feces/virology , Genome, Viral , Humans , Infant , Microscopy, Electron , Molecular Sequence Data , Orthoreovirus/isolation & purification , Orthoreovirus, Mammalian/genetics , Phylogeny , RNA, Viral/genetics , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Sequence Homology, Nucleic Acid , Slovenia , Virus Cultivation
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