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디스크확산법에 의한 Helicobacter pylori의 Clarithromycin과 Amoxicillin 항균제감수성 검사

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

이종욱|이혜경|임지훈|김미나|이미경|이미애|강정옥|성흥섭|Helicobacter 연구회
대한임상미생물학회 (Korean Society of Clinical Microbiology)|2009.03|Annals of Clinical Microbiology|제12권 제1호|pp. 30-36

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키워드

Helicobacter pyloriClarithromycinAgar dilutionDisk diffusionAmoxicillin

초록

배경: Helicobacter pylori 항균제감수성 검사는 clarithromycin에 대한 한천희석법만 Clinical and Laboratory Standard Institute (CLSI) 지침이 정해져 있다. 본 연구에서는 임상검사실에서 H. pylori의 clarithromycin과 amoxicillin 항균제감수성 검사를 위해 디스크확산법을 평가하고자 하였다. 방법: 2005년 5월부터 2007년 5월까지 임상검체에서 분리된 140균주의 H. pylori를 대상으로 하였다. CLSI 지침에 따라 한천희석법으로 clarithromycin과 amoxicillin에 대한 최소억제농도(minimum inhibition concentration, MIC)를 구했으며, 디 스크확산법은 2μg과 15μg의 clarithromycin, 2μg과 10μg의 amoxicillin 디스크를 사용하여 5% 면양혈액 Mueller-Hinton 한천배지에서 검사하였다. 최소억제농도와 디스크 억제대를 선형회귀분석과 error rate-bounded 방법으로 비교하여 디스 크확산법의 감수성 판독 기준을 설정하였다. 결과: 한천희석법 MIC 결과 clarithromycin 내성과 중간내성의 비율은 각각 21.4%, 1.4%였으며, amoxicillin MIC가 0.5, 1, 2μg/mL인 균주는 각각 7주, 2주, 1주였다. 디스크확산법에서 clarithromycin 2μg과 15μg 디스크 억제대는 log2 MIC와 R2값이 각각 0.931, 0.923이었으며(P＜0.001), 내성/중간내성 기준은 각각 12/28 mm와 23/39 mm로 설정하였다. Amoxicillin 2μg와 10μg 디스크 억제대는 log2 MIC와 R2값이 각각 0.478과 0.421이었으며(P＜0.001), 억제대의 판독기준은 MIC 내성 기준을 2μg/mL로 하면 21 mm와 33 mm였다. 2μg 디스크에서는 억제대 지름이 모두 60 mm 미만인데 비해 15μg clarithromycin 디스크에서는 61.4% (86/140), 10μg amoxicillin 디스크에서는 75.7% (106/140)가 억제대 지름이 60 mm 이상이 었다. 결론: Clarithromycin, amoxicillin 디스크확산법은 억제대의 지름이 MIC와 높은 상관성을 보였으며, MIC에 의한 감수성 판정과 100% 일치하는 억제대 지름의 판정기준을 설정할 수 있었다. 2μg 디스크의 감수성 판정기준이 28 mm 이하로 더 적절하였으며 두 디스크를 하나의 평판배지에서 검사할 수 있었다.

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.

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