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Journal of Clinical Microbiology, September 2001, p. 3052-3055, Vol. 39, No. 9
Department of Medical Microbiology, Royal Free and
University College Medical School,1 and
Helicobacter Reference Unit, Central Public Health
Laboratory,2 London, and Institute of
Infections and Immunity, University of Nottingham,
Nottingham,4 United Kingdom, and
Unité de Pathogénie Bactérienne des
Muqueuses, Institut Pasteur, Paris, France3
Received 27 March 2001/Returned for modification 23 April
2001/Accepted 14 June 2001
Antimicrobial resistance in Helicobacter pylori is a
serious and increasing problem, and the development of rapid, reliable methods for detecting resistance would greatly improve the selection of
antibiotics used to treat gastric infection with this organism. We
assessed whether detection of the RdxA protein could provide the basis
for determining the susceptibility of H. pylori to
metronidazole. In order to raise polyclonal antisera to RdxA, we cloned
the rdxA gene from H. pylori strain 26695 into
the commercial expression vector pMAL-c2, purified the resultant fusion
protein by affinity chromatography, and used this recombinant RdxA
preparation to immunize rabbits. We then used this specific anti-RdxA
antibody to perform immunoblotting on whole bacterial cell lysates of
17 metronidazole-sensitive and 27 metronidazole-resistant clinical isolates of H. pylori. While a 24-kDa immunoreactive band
corresponding to the RdxA protein was observed in all
metronidazole-sensitive strains, this band was absent in 25 of 27 resistant isolates. Our results indicate that testing for the absence
of the RdxA protein would identify the majority of clinical isolates
that will respond poorly to metronidazole-containing eradication
regimens and have implications for the development of assays capable of detecting metronidazole resistance in H. pylori.
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.9.3052-3055.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Differentiation of Metronidazole-Sensitive and -Resistant
Clinical Isolates of Helicobacter pylori by
Immunoblotting with Antisera to the RdxA Protein
*
Corresponding author. Mailing address: Institute of
Infections and Immunity, Floor C, West Block, University Hospital,
Queen's Medical Centre, Nottingham NG7 2UH, United Kingdom. Phone:
44-115-9249924, ext. 42457. Fax: 44-115-9709923. E-mail:
Peter.Jenks{at}nottingham.ac.uk.
Journal of Clinical Microbiology, September 2001, p. 3052-3055, Vol. 39, No. 9
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.9.3052-3055.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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