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Sung, Kang, Lee, Lee, Lee, Lee, Lim, Kim, and Helicobacter Study Group: Clarithromycin and Amoxicillin Susceptibility Testing of Helicobacter pylori by Disk Diffusion Method
Original Article

Korean J Clin Microbiol 2009;12(1):30-36.

Published online: 22 March 2009

DOI: https://doi.org/10.5145/KJCM.2009.12.1.30

Clarithromycin and Amoxicillin Susceptibility Testing of Helicobacter pylori by Disk Diffusion Method

Heungsup Sung1, Jung-Oak Kang2, Mi Ae Lee3, Jongwook Lee4, Hae Kyung Lee5, Mi-Kyung Lee6, Ji-Hun Lim1, Mi-Na Kim1; Helicobacter Study Group

1Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea

2Hanyang University Medical College, Seoul, Korea

3Ewha Womans University College of Medicine, Seoul, Korea

4Konyang University College of Medicine, Seoul, Korea

5The Catholic University of Korea College of Medicine, Seoul, Korea

6Chung-Ang University College of Medicine, Seoul, Korea

Correspondence: Mi-Na Kim, Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, 388-1, Pungnap-2dong, Songpa-gu, Seoul 138-736, Korea. (Tel) 82-2-3010-4511, (Fax) 82-2-478-0884, (E-mail) mnkim@amc.seoul.kr

Received 10 November 20089 December 2008       Accepted 20 January 2009

Copyright © 2009 Korean Journal of Clinical Microbiology

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

CLSI provides a guideline only for a agar dilution method of testing clarithromycin susceptibility for Helicobacter pylori. This study was to evaluate a disk diffusion method for clarithromycin and amoxicillin.

Methods

One hundred and forty clinical isolates of H. pylori isolated from May 2005 to May 2007 were tested by the CLSI agar dilution method and a disk diffusion method using 2μg (2CLR) and 15μg (15CLR) clarithromycin disks and 2μg (2AMX) and 10μg (10AMX) amoxicillin disks. The interpretation criteria used for the disk diffusion method were established by linear regression and error rate-bounded method for disk diffusion zone of inhibition (DDZ) compared to MIC.

Results

Resistance and intermediate rates to clarithromycin were 21.4% and 1.4%, respectively. A number of isolates with MIC 0.5, 1, and 2 (μg/mL) to amoxicillin were 7, 2, and 1, respectively. For 2CLR and 15CLR, the coefficients of determination (R2) between MIC and DDZ were 0.931 and 0.923 (P＜ 0.001), respectively, and the criteria for resistance/ susceptibility were 12/28 mm for 2CLR and 23/39 mm for 15CLR. For 2AMX and 10AMX, the R2 between MIC and DDZ were 0.478 and 0.421 (P＜ 0.001), respectively, and the criteria for resistance with breakpoint of 2μg/mL were 21 mm for 2AMX and 32 mm for 10AMX. All isolates had DDZ＜60 mm with 2CLR and 2AMX, but 61.4% and 75.7% of the isolates had DDZ＜60 mm with 15CLR and 10AMX, respectively.

Conclusion

Excellent correlation and agreement between MIC and DDZ were found for clarithromycin and amoxicillin. With 2μg disks, the susceptibility breakpoints were 28 mm or less; thus, two disks could be tested in one plate.

Keywords: Helicobacter pylori, Agar dilution, Disk diffusion, Clarithromycin, Amoxicillin

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Fig. 1.
Error rate-bounded analyses of agar dilution MIC versus disk diffusion zone diameter for clarithromycin against 140 H. pylori clinical isolates. There was no major or minor error with resistance and susceptibility criteria of 12 mm and 28 mm for 2μg disk (A) and 23 mm and 34 mm for 15μg disk (B). The number of isolates more than 10 was indicated with underline.
kjcm-12-30f1.tif
Fig. 2.
Error rate-bounded analyses of agar dilution MIC versus disk diffusion zone diameter for amoxicillin against 140 H. pylori clinical isolates. Dotted line and dotdash lines were for the breakpoint of 2μg/mL and 0.5μg/mL, respectively. At the resistance breakpoint of 2μg/mL, there was no major or minor error when 21 mm for 2μg disk (A) and 32 mm for 10μg disk (B) was applied for resistance criteria. The number of isolates more than 10 was indicated with underline.
kjcm-12-30f2.tif
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