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Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca와 Proteus mirabilis 에서 Boronic Acid Disk를 이용한 Extended-spectrum β-lactamase와 AmpC β-lactamase 검출

Use of Boronic Acid Disks for the Detection of Extended-spectrum β-lactamase and AmpC β-lactamase in Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca and Proteus mirabilis

장인호|어영|윤갑준|김효열|박순덕|권오건
대한임상미생물학회 (Korean Society of Clinical Microbiology)|2009.03|Annals of Clinical Microbiology|제12권 제1호|pp. 24-29

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키워드

Extended-spectrum β-lactamasePlasmidmediated AmpC β-lactamaseBoronic acidCoproducers

초록

배경: Extended spectrum β-lactamase (ESBL)와 AmpC β-lactamase를 생성하는 균종의 정확한 검출은 환자의 치료에 매우 중요하다. 그러나 ESBL 검출법과는 달리 AmpC β-lactamase는 표준화된 검출 방법이 없다. 본 연구에서는 boronic acid를 이용한 ESBL과 AmpC β-lactamase 검출법을 평가하였다. 방법: Cefoxitin 중간내성이거나 내성 또는 ESBL 시험에서 양성을 보인 Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca와 Proteus mirabilis 균종을 대상으로 하였다. ESBL 생성은 ceftazidime/clavulanic acid/boronic acid (CAZ/CA/BA)의 억제대 지름에서 CAZ/BA의 억제대 지름을 뺀 값이 5 mm 이상이거나 cefotaxime/clavulanic acid/boronic acid (CTX/CA/BA)의 억제대 지름에서 CTX/BA의 억제대 지름을 뺀 값이 5 mm 이상이면 ESBL로 판정하고, cefoxitin/boronic acid (FOX/BA)의 억제대 지름에서 FOX의 억제대 지름을 뺀 값이 5 mm 이상이거나 cefotetan/boronic acid (CTT/BA)의 억제대 지름에서 CTT의 억제대 지름을 뺀 값이 5 mm 이상이면 AmpC β-lactamase 생성 균주로 판정하였다. 결과: 총 622 균주로 CLSI ESBL 확진검사를 시행한 결과, 균종별 ESBL 양성률은 E. coli 18.1%, K. pneumoniae 38.3%, K. oxytoca 8.7%, P. mirabilis 14.8%였고, boronic acid를 이용한 ESBL 검출법으로 측정한 ESBL 양성률은 E. coli 18.4%, K. pneumoniae 40.4%, K. oxytoca 8.7%, P. mirabilis 14.8%였다. Boronic acid를 이용한 AmpC 검출 양성률은 E. coli 3.7%, K. pneumoniae 33.3%, K. oxytoca 0%, P. mirabilis 7.4%였고 균종별로 ESBL과 AmpC를 동시에 생성하는 균의 비율은 E. coli 2.4%, K. pneumoniae 27.1%, P. mirabilis 3.7%였다. 결론: Boronic acid를 이용한 ESBL 확진법은 ESBLs 검출력을 높일 수 있었고 AmpC에 의해 가려져 있는 ESBLs도 검출할 수 있었다.

Abstract

Background: Accurate detection of organisms producing extended-spectrum β-lactamase (ESBL) and AmpC β-lactamase is very important for treatment of patients. However, unlike the ESBL confirmatory test, there are no guidelines for detection of organisms producing AmpC β-lactamase. We evaluated a detection method using boronic acid (BA) for ESBL and AmpC β-lactamase. Methods: Clinical isolates of Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, and Proteus mirabilis showing intermediate resistance or resistance to cefoxitin (FOX) or positive for ESBL were tested. A ≥5 mm increase in zone diameter of ceftazidime/ clavulanic acid/BA (CAZ/CA/BA) and/or cefotaxime/ clavulanic acid/BA (CTX/CA/BA) versus CAZ/BA and/or CTX /BA was considered positive for ESBL. Likewise, a ≥5 mm increase in zone diameter of FOX/BA and/or cefotetan/BA (CTT/BA) versus FOX and/or CTT alone was considered positive for AmpC β-lactamase. Results: Among 622 clinical isolates, ESBL positive rates by the CLSI ESBL confirmatory test or by the BA method were 18.1% or 18.4% for E. coli, 38.3% or 40.4% for K. pneumoniae, 8.7% or 8.7% for K. oxytoca, and 14.8% or 14.8% for P. mirabilis, respectively. AmpC β-lactamase positive rates using the BA method were 3.7% for E. coli, 33.3% for K. pneumoniae, 0% for K. oxytoca, and 7.4% for P. mirabilis. The detection rates of coproducing ESBL and AmpC β-lactamase were 2.4% in E. coli 27.1% in K. pneumoniae, and 3.7% in P. mirabilis. Conclusion: The ESBL confirmatory method using BA was found to enhance the detection of ESBLs, even when potentially masked by AmpC β-lactamase.

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