Document Detail


A rapid and sensitive procedure for determination of 5-N-acetyl neuraminic acid in lipopolysaccharides of Haemophilus influenzae: a survey of 24 non-typeable H. influenzae strains.
MedLine Citation:
PMID:  11595219     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In view of the importance of 5-N-acetyl neuraminic acid in bacterial pathogenesis, a sensitive, reproducible and reliable method for the determination of 5-N-acetyl neuraminic acid levels in lipopolysaccharide (LPS) is described and applied to 24 different non-typeable Haemophilus influenzae (NTHi) strains. The method involves analysis by high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) of terminal 5-N-acetyl neuraminic acid residues released by neuraminidase treatment of O-deacylated LPS. The procedure is relatively fast and the instrumental effort is moderate. The results of the procedure were compared with data obtained by 1H NMR and electrospray ionisation-mass spectrometry (ESI-MS). The analysis of LPS from 24 NTHi strains showed that 5-N-acetyl neuraminic acid was found to be a common constituent of LPS in NTHi. Only one strain (NTHi 432) did not show any sialylation. Molar ratios (LPS/5-N-acetyl neuraminic acid) ranged between 5/1 and 500/1. Several strains in which no 5-N-acetyl neuraminic acid could be determined by other methods including 1H NMR and ESI-MS were shown to contain 5-N-acetyl neuraminic acid by this HPAEC-PAD procedure. The method was applied to determine levels of terminal 5-N-acetyl neuraminic acid in LPS from NTHi strains grown under different conditions and mutant strains containing inactive LPS biosynthetic genes.
Authors:
S H Bauer; M Månsson; D W Hood; J C Richards; E R Moxon; E K Schweda
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Carbohydrate research     Volume:  335     ISSN:  0008-6215     ISO Abbreviation:  Carbohydr. Res.     Publication Date:  2001 Oct 
Date Detail:
Created Date:  2001-10-11     Completed Date:  2002-01-29     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0043535     Medline TA:  Carbohydr Res     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  251-60     Citation Subset:  IM    
Affiliation:
Clinical Research Centre, Karolinska Institutet and University College of South Stockholm, NOVUM, S-141 86 Huddinge, Sweden.
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MeSH Terms
Descriptor/Qualifier:
Chromatography, Ion Exchange / methods,  standards
Haemophilus influenzae / chemistry*,  classification
Lipopolysaccharides / chemistry*,  metabolism
Magnetic Resonance Spectroscopy
N-Acetylneuraminic Acid / analysis*
Neuraminidase / metabolism
Reproducibility of Results
Sensitivity and Specificity
Spectrometry, Mass, Electrospray Ionization
Chemical
Reg. No./Substance:
0/Lipopolysaccharides; 131-48-6/N-Acetylneuraminic Acid; EC 3.2.1.18/Neuraminidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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