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J. Clin. Microbiol. doi:10.1128/JCM.00681-08
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Rapid method for sensitive screening of oligosaccharide epitopes in the lipooligosaccharide from Campylobacter jejuni strains isolated from Guillain-Barré syndrome and Miller Fisher syndrome patients

Institute for Biological Sciences, National Research Council Canada, Ottawa, Ontario, Canada, K1A 0R6; Department of Medical Microbiology and Infectious Diseases, Erasmus University Medical Center Rotterdam, The Netherlands; Clinical Research Centre, Karolinska Institute and University College of South Stockholm, NOVUM, S-141 86 Huddinge, Sweden

^* To whom correspondence should be addressed. Email: Jianjun.Li{at}nrc-cnrc.gc.ca.

	Abstract

Campylobacter jejuni lipooligosaccharide (LOS) can trigger Guillain-Barré syndrome (GBS) due to its similarity with human gangliosides. Rapid and accurate structural elucidation of the LOS glycan of a strain isolated from a GBS patient could help physicians determine the spectrum of anti-ganglioside antibodies likely to be found, and therefore provide valuable assistance in establishing an appropriate course of treatment. The ability of implement mass spectrometry-based approaches in a clinical setting have been limited by the laborious and time consuming nature of the protocols, typically 3 to 4 days, used to prepare LOS. In order to improve the analytical throughput, microwave-assisted enzymatic digestion was investigated. In this study, the bacterial cells were suspended in 50 µL of 20 mM ammonium acetate buffer containing DNase and RNase and treated by direct microwave irradiation for 3 minutes. Then, proteinase K was added and the samples were again microwaved. The intact LOS samples were analysed using electrophoresis-assisted open-tubular liquid chromatography-mass spectrometry. The reliability of the rapid high-throughput technique was demonstrated through the analysis of the LOS glycan from 73 C. jejuni strains. The structure could be elucidated using material from a single colony. The total time for sample preparation and MS analysis is less than 60 min.