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- Title
MS-H: A Novel Proteomic Approach to Isolate and Type the <i>E. coli</i> H Antigen Using Membrane Filtration and Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS).
- Authors
Cheng, Keding; Drebot, Mike; McCrea, Joanne; Peterson, Lorea; Lee, David; McCorrister, Stuart; Nickel, Richard; Gerbasi, Alyssia; Sloan, Angela; Janella, Debra; Van Domselaar, Gary; Beniac, Daniel; Booth, Tim; Chui, Linda; Tabor, Helen; Westmacott, Garrett; Gilmour, Matthew; Wang, Gehua
- Abstract
Serotyping is the long-standing gold standard method to determine E. coli H antigens; however, this method requires a panel of H-antigen specific antibodies and often culture-based induction of the H-antigen flagellar motility. In this study, a rapid and accurate method to isolate and identify the Escherichia coli (E. coli) H flagellar antigen was developed using membrane filtration and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Flagella were isolated from pure culture, digested with trypsin, and then subjected to LC-MS/MS using one of two systems (Agilent-nano-LC-QSTAR XL or Proxeon-nano-LC-LTQ-Orbitrap XL). The resulting peptide sequence data were searched against a custom E. coli flagella/H antigen database. This approach was evaluated using flagella isolated from reference E. coli strains representing all 53 known H antigen types and 41 clinical E. coli strains. The resulting LC-MS/MS classifications of H antigen types (MS-H) were concordant with the known H serogroup for all 53 reference types, and of 41 clinical isolates tested, 38 (92.7%) were concordant with the known H serogroup. MS-H clearly also identified two clinical isolates (4.9%) that were untypeable by serotyping. Notably, successful detection and classification of flagellar antigens with MS-H did not generally require induction of motility, establishing this proteomic approach as more rapid and cost-effective than traditional methods, while providing equitable specificity for typing E. coli H antigens.
- Subjects
PROTEOMICS; ESCHERICHIA coli; BACTERIAL antigens; MEMBRANE separation; LIQUID chromatography; TANDEM mass spectrometry; SEROTYPING
- Publication
PLoS ONE, 2013, Vol 8, Issue 2, p1
- ISSN
1932-6203
- Publication type
Article
- DOI
10.1371/journal.pone.0057339