MS-H: A novel proteomic approach to isolate and type the E. coli H antigen using membrane filtration and liquid chromatography-tandem mass spectrometry (LC-MS/MS)

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 and Wang, Gehua (2013) MS-H: A novel proteomic approach to isolate and type the E. coli H antigen using membrane filtration and liquid chromatography-tandem mass spectrometry (LC-MS/MS). PLoS One, 8 (2). ISSN 1932-6203

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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.

Item Type:	Article
Uncontrolled Keywords:	biochemistry, genetics and molecular biology(all),agricultural and biological sciences(all),general ,/dk/atira/pure/subjectarea/asjc/1300
Faculty \ School:	Faculty of Medicine and Health Sciences > Norwich Medical School
Related URLs:	http://www.scopus.com/inward/record.url?...
Depositing User:	LivePure Connector
Date Deposited:	14 Sep 2022 09:30
Last Modified:	31 Oct 2023 02:35
URI:	https://ueaeprints.uea.ac.uk/id/eprint/88237
DOI:	10.1371/journal.pone.0057339

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