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Journal of Clinical Microbiology, October 2008, p. 3429-3436, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.00681-08
Copyright © 2008, American Society for Microbiology. 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 ^,

Monika Dzieciatkowska,¹ Xin Liu,¹ Astrid P. Heikema,² R. Scott Houliston,¹ Alex van Belkum,² Elke K. H. Schweda,³ Michel Gilbert,¹ James C. Richards,¹ and Jianjun Li^1*

Institute for Biological Sciences, National Research Council Canada, Ottawa, Ontario, Canada K1A 0R6,¹ Department of Medical Microbiology and Infectious Diseases, Erasmus University Medical Center Rotterdam, Rotterdam, The Netherlands,² Clinical Research Centre, Karolinska Institute and University College of South Stockholm, NOVUM, S-141 86 Huddinge, Sweden³

Received 10 April 2008/ Returned for modification 30 July 2008/ Accepted 19 August 2008

Campylobacter jejuni lipooligosaccharide (LOS) can trigger Guillain-Barré syndrome (GBS) due to its similarity to 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 implemented mass spectrometry-based approaches in a clinical setting has 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 min. Then, proteinase K was added and the samples were again microwaved. The intact LOS samples were analyzed using electrophoresis-assisted open-tubular liquid chromatography-mass spectrometry. The reliability of the rapid, high-throughput technique was demonstrated through analysis of LOS glycans from 73 C. jejuni strains. The structure was elucidated using material from a single colony. The total time for sample preparation and MS analysis is less than 60 min.

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* Corresponding author. Mailing address: NRC Institute for Biological Sciences, National Research Council Canada, 100 Sussex Drive, Ottawa, Ontario, Canada K1A 0R6. Phone: (613) 990-0558. Fax: (613) 952-9092. E-mail: Jianjun.Li{at}nrc-cnrc.gc.ca

Published ahead of print on 27 August 2008.

Supplemental material for this article may be found at http://jcm.asm.org/.

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Journal of Clinical Microbiology, October 2008, p. 3429-3436, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.00681-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

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