Phenotypic H-Antigen Typing by Mass Spectrometry Combined with Genetic Typing of H Antigens, O Antigens, and Toxins by Whole-Genome Sequencing Enhances Identification of Escherichia coli Isolates

نویسندگان

  • Keding Cheng
  • Huixia Chui
  • Larissa Domish
  • Angela Sloan
  • Drexler Hernandez
  • Stuart McCorrister
  • Alyssia Robinson
  • Matthew Walker
  • Lorea A. M. Peterson
  • Miles Majcher
  • Sam Ratnam
  • David J. M. Haldane
  • Sadjia Bekal
  • John Wylie
  • Linda Chui
  • Shaun Tyler
  • Bianli Xu
  • Aleisha Reimer
  • Celine Nadon
  • J. David Knox
  • Gehua Wang
چکیده

Mass spectrometry-based phenotypic H-antigen typing (MS-H) combined with whole-genome-sequencing-based genetic identification of H antigens, O antigens, and toxins (WGS-HOT) was used to type 60 clinical Escherichia coli isolates, 43 of which were previously identified as nonmotile, H type undetermined, or O rough by serotyping or having shown discordant MS-H and serotyping results. Whole-genome sequencing confirmed that MS-H was able to provide more accurate data regarding H antigen expression than serotyping. Further, enhanced and more confident O antigen identification resulted from gene cluster based typing in combination with conventional typing based on the gene pair comprising wzx and wzy and that comprising wzm and wzt The O antigen was identified in 94.6% of the isolates when the two genetic O typing approaches (gene pair and gene cluster) were used in conjunction, in comparison to 78.6% when the gene pair database was used alone. In addition, 98.2% of the isolates showed the existence of genes for various toxins and/or virulence factors, among which verotoxins (Shiga toxin 1 and/or Shiga toxin 2) were 100% concordant with conventional PCR based testing results. With more applications of mass spectrometry and whole-genome sequencing in clinical microbiology laboratories, this combined phenotypic and genetic typing platform (MS-H plus WGS-HOT) should be ideal for pathogenic E. coli typing.

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عنوان ژورنال:

دوره 54  شماره 

صفحات  -

تاریخ انتشار 2016