Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: infection due to Escherichia coli [ICD11:
XN6P4 
]
NCBI PubMed ID: 24789798Publication DOI: 10.1128/CVI.00685-13Journal NLM ID: 101252125Publisher: American Society for Microbiology
Correspondence: Gábor Nagy <gabor.nagy
arsanis.com>
Institutions: Arsanis Biosciences GmbH, Vienna, Austria, Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland
The Escherichia coli lineage ST131-O25b:H4 is a globally spread multi-drug resistant clone responsible for a significant proportion of extraintestinal infections. Driven by the high medical need associated with this successful pathogenic lineage, we generated murine monoclonal antibodies against its lipopolysaccharide (LPS) O25b antigen in order to develop quick diagnostic tests. Murine mAbs were generated by immunization of mice with whole killed non-encapsulated ST131-O25b E. coli cells and screening hybridoma supernatants for binding to purified LPS molecules obtained from an E. coli ST131-O25b clinical isolate. The mAbs selected for further study bound to the surface of live E. coli O25b strains irrespective of the capsular type expressed, while they could not bind to bacteria or purified LPS from other serotypes - including the related classical O25 antigen (O25a). Using these specific mAbs we have developed a latex bead-based agglutination assay that has greater specificity, more rapid and simpler than the currently available typing methods. The high specificity of these mAbs can be explained by the novel structure of the O25b repeating unit elucidated in this paper. Based on comparative analysis by NMR and mass spectrometry, the N-acetyl-fucose in the O25a O-antigen had been replaced by O-acetyl-rhamnose in the O25b repeating unit. The genetic determinants responsible for this structural variation were identified by alignment of corresponding genetic loci, and were confirmed by trans-complementation of a rough mutant by the sub-serotype specific fragments of the rfb operons.
O-antigen, Escherichia coli, antibodies, monoclonal antibodies, MAb, Serotypes, binding, typing methods
Structure type: polymer chemical repeating unit
Location inside paper: p.933, p.935, fig.4A
Compound class: O-polysaccharide, O-antigen
Contained glycoepitopes: IEDB_135813,IEDB_136105,IEDB_137340,IEDB_141806,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_151531,IEDB_225177,IEDB_885823,IEDB_983931,SB_192
Methods: 13C NMR, 1H NMR, methylation, NMR-2D, GC-MS, SDS-PAGE, DNA techniques, ELISA, 31P NMR, MALDI-TOF MS, composition analysis, NMR-1D, serological methods, genetic methods, immunoblotting
Related record ID(s): 30515, 30516, 30517
NCBI Taxonomy refs (TaxIDs): 1095709Reference(s) to other database(s): GTC:G18223OD, GlycomeDB:
28106
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There is only one chemically distinct structure:
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