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In Silico Genotyping of Escherichia coli Isolates for Extraintestinal Virulence Genes by Use of Whole-Genome Sequencing Data

Extraintestinal pathogenic (ExPEC) is the leading cause in humans of urinary tract infection and bacteremia. The previously published web tool VirulenceFinder (http://cge.cbs.dtu.dk/services/VirulenceFinder/) uses whole-genome sequencing (WGS) data for characterization of isolates and enables resear...

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Bibliographic Details
Published in:Journal of clinical microbiology 2020-09, Vol.58 (10)
Main Authors: Malberg Tetzschner, Anna Maria, Johnson, James R, Johnston, Brian D, Lund, Ole, Scheutz, Flemming
Format: Article
Language:English
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Summary:Extraintestinal pathogenic (ExPEC) is the leading cause in humans of urinary tract infection and bacteremia. The previously published web tool VirulenceFinder (http://cge.cbs.dtu.dk/services/VirulenceFinder/) uses whole-genome sequencing (WGS) data for characterization of isolates and enables researchers and clinical health personnel to quickly extract and interpret virulence-relevant information from WGS data. In this study, 38 ExPEC-associated virulence genes were added to the existing VirulenceFinder database. In total, 14,441 alleles were downloaded. A total of 1,890 distinct alleles were added to the database after removal of redundant sequences and analysis of the remaining alleles for open reading frames (ORFs). The database now contains 139 genes-of which 44 are related to ExPEC-and 2,826 corresponding alleles. Construction of the database included validation against 27 primer pairs from previous studies, a search for serotype-specific P fimbriae alleles, and a BLASTn confirmation of seven genes ( , , , , , , and ) not covered by the primers. The augmented database was evaluated using (i) a panel of nine control strains and (ii) 288 human-source strains classified by PCR as ExPEC and non-ExPEC. We observed very high concordance (average, 93.4%) between PCR and WGS findings, but WGS identified more alleles. In conclusion, the addition of 38 ExPEC-associated genes and the associated alleles to the VirulenceFinder database allows for a more complete characterization of isolates based on WGS data, which has become increasingly important considering the plasticity of the genome.
ISSN:0095-1137
1098-660X
DOI:10.1128/JCM.01269-20