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Journal of Clinical Microbiology, April 2001, p. 1217-1220, Vol. 39, No. 4
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.4.1217-1220.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

PCR-Based Diagnosis of Helicobacter pylori Infection and Real-Time Determination of Clarithromycin Resistance Directly from Human Gastric Biopsy Samples

Stephanie A. Chisholm,1 Robert J. Owen,1,* E. Louise Teare,2 and Seth Saverymuttu3

Laboratory of Enteric Pathogens, Central Public Health Laboratory, London NW9 5HT,1 Public Health Laboratory, Chelmsford CM2 OYX,2 and Broomfield Hospital, Chelmsford CM1 7ET,3 United Kingdom

Received 6 September 2000/Returned for modification 20 November 2000/Accepted 12 January 2001

A novel PCR detection assay that amplifies the Helicobacter pylori-specific vacuolating cytotoxin gene (vacA) and thus enables rapid diagnosis of infection is described. Additionally, a real-time probe hybridization melting point analysis assay to detect all three mutations in the 23S rRNA gene associated with clarithromycin resistance was applied directly to antral gastric biopsy samples. Comparison with culture and an alternative PCR assay targeting the 16S rrn gene showed that the vacA assay was sensitive and specific when tested on biopsy samples from 121 patients. Clarithromycin susceptibilities could be determined in the majority (92.3%) of culture-positive gastric biopsy samples analyzed, four of which generated melting peaks indicative of clarithromycin resistance by either an Aright-arrowG or Aright-arrowC mutation. The presence of the mutations correlated with the clarithromycin disk diffusion sensitivities of matched cultures. This PCR-based system was simple to perform and could be completed in 3 to 4 h, thereby overcoming the delays associated with conventional culture methods for H. pylori identification and susceptibility testing.

* Corresponding author. Mailing address: Helicobacter Reference Unit, Laboratory of Enteric Pathogens, Central Public Health Laboratory, 61 Colindale Ave., Colindale, London NW9 5HT, United Kingdom. Phone: (44) 20 8200 4400. Fax: (44) 20 8905 9929. E-mail: rowen{at}phls.nhs.uk.

Journal of Clinical Microbiology, April 2001, p. 1217-1220, Vol. 39, No. 4
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.4.1217-1220.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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