Se Jin Oh; Sang Uk Lee; Hyun Yong Hwang; Il Kwon Bae; Hyun Soo Jo; Byung Ho Lee; Seok Hoon Jeong

doi:10.3343/kjlm.2006.26.1.14

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Oh, Lee, Hwang, Bae, Jo, Lee, and Jeong: Prevalence of Class A Extended-Spectrum β-Lactamases in Clinical Isolates of Acinetobacter baumannii and Pseudomonas aeruginosa

Original Article

Korean J Leg Med 2006;26(1):14-20.

Published online: 10 February 2006

DOI: https://doi.org/10.3343/kjlm.2006.26.1.14

Prevalence of Class A Extended-Spectrum β-Lactamases in Clinical Isolates of Acinetobacter baumannii and Pseudomonas aeruginosa

Se Jin Oh, M.D., Sang Uk Lee, M.D., Hyun Yong Hwang, M.D.¹, Il Kwon Bae, M.D.¹, Hyun Soo Jo, M.D.¹, Byung Ho Lee, M.D.¹, Seok Hoon Jeong, M.D.¹

Department of Internal Medicine, Kosin University College of Medicine, Busan, Korea

¹Department of Laboratory Medicine, Kosin University College of Medicine, Busan, Korea

Received 22 September 2005 Revised 6 January 2006 Accepted 13 January 2006

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

Prevalence of class A extended-spectrum β-lactamases (ESBLs) has been investigated repeatedly in members of family Enterobacteriaceae in Korea, but only rarely in Acinetobacter baumannii and Pseudomonas aeruginosa. The aims of this study were to determine the prevalence of class A ESBL-producing A. baumannii and P. aeruginosa and to characterize the genotypes.

Methods

During the period of June to September 2004, clinical isolates of A. baumannii and P. aeruginosa were collected from patients in Kosin University Gospel Hospital, Busan, Korea. Antimicrobial susceptibility was determined by the disk diffusion and the agar dilution methods, and ESBL-production by the double-disk synergy test. Transferability of ceftazidime-resistance of ESBL-producers were tested by conjugation. The isoelectric points of ESBLs were determined by isoelectric focusing. Searches for blaTEM, blaSHV, blaCTX-M, blaPER-1, blaVEB, and blaGES/IBC genes were performed by PCR amplification, and the genotypes of ESBLs were determined by a direct nucleotide sequence analysis of the amplified products.

Results

A total of 58 clinical isolates of A. baumannii and 77 P. aeruginosa were collected. Three (5.2%) isolates of A. baumannii and four (5.2%) P. aeruginosa isolates showed positive results in the double-disk synergy test using ceftazidime and imipenem disks, and one (1.7%) A. baumannii and two (2.6%) P. aeruginosa isolates showed positive results in that test using ceftazidime and cefoxitin disks. The most prevalent class A ESBL genotype in A. baumannii isolates was blaPER-1 (n=6), and blaSHV-12 gene was also found in one P. aeruginosa isolate.

Conclusions

It is concluded that class A PER-1 ESBL-producing A. baumannii isolates are spreading, and SHV-12-producing P. aeruginosa has emerged in Korea. The spread of class A ESBLs could compromise the future usefulness of expanded-spectrum β-lactam antibiotics for the treatment of A. baumannii and P. aeruginosa infections.

Keywords: Acinetobacter baumannii, Pseudomonas aeruginosa, Class A ESBL

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Fig. 1.

An example of double disk synergy obtained with an A baumannii isolate producing Ambler class A extended-spectrum β-lactamase. Two 30 μg-antibiotics disks (left, imipenem; right, cefoxitin) were placed 1.5 cm apart (margin to margin) around a disk containing 30 μg of ceftazidime.

Table 1.

Antimicrobial susceptibilities of A. baumannii and P. aeruginosa isolates

Antimicrobial agents	% Susceptibility
	A. baumannii (n=58)		P. aeruginosa (n=77)
	Intermediate	Resistant	Intermediate	Resistant
Ampicillin	3.4	94.9	ND	ND
Ampicillin-sulbactam	3.5	14.6	ND	ND
Piperacillin	7.0	24.6	0.0	39.0
Piperacillin-tazobactam	0.0	21.1	0.0	37.8
Cefoxitin	7.3	87.2	ND	ND
Cefotetan	3.6	94.6	ND	ND
Ceftazidime	5.2	25.8	5.2	29.9
Cefotaxime	60.3	32.7	36.9	58.7
Cefepime	5.2	22.4	10.0	26.0
Aztreonam	27.6	67.2	20.8	29.9
Imipenem	0.0	24.1	13.0	31.2
Meropenem	0.0	24.1	3.9	40.3
Amikacin	0.0	27.6	2.6	29.6
Gentamicin	0.0	29.4	2.6	46.0
Tobramycin	0.0	29.8	0.0	44.2
Ciprofloxacin	1.7	24.0	ND	ND
Trimethoprim-sulfamethoxazole	7.0	26.3	ND	ND
Tetracycline	0.0	23.6	ND	ND

Abbreviations: ND, not done.

Table 2.

Characteristics of double-disk synergy-positive A. baumannii and P. aeruginosa isolates

DDS (+) with	Isolate	Species	Specimen	MICs (μg/mL)
DDS (+) with	Isolate	Species	Specimen	PIP	TZP	CAZ	CAZ/CLA^*	FEP
Imipenem disk	K20653	A. baumannii	Sputum	256	32	128	256	32
	K22565	A. baumannii	Sputum	16	4	8	8	4
	K22898	A. baumannii	Urine	4	1	2	2	1
	K18885	P. aeruginosa	Urine	32	32	128	256	32
	K19135	P. aeruginosa	Urine	16	8	128	256	32
	K24002	P. aeruginosa	Urine	32	8	128	256	32
	K24857	P. aeruginosa	Pus	64	32	>256	>256	128
Cefoxitin disk	K24931	A. baumannii	Sputum	16	4	32	32	16
	K23180	P. aeruginosa	Sputum	1	1	1	2	8
	K23413	P. aeruginosa	Sputum	8	4	2	4	4

^* clavulanic acid at a fixed concentration of 4 μg/mL.

Abbreviations: DDS (+), double-disk synergy-positive; PIP, piperacillin; TZP, piperacillin-tazobactam; CAZ, ceftazidime; CAZ/CLA, ceftazidime-clavulanic acid; FEP, cefepime.

Table 3.

Characteristics of TEM-116-producing A. baumannii and P. aeruginosa isolates

Isolate	Species	Specimen	MICs (μg/mL)
Isolate	Species	Specimen	PIP	TZP	CAZ	CAZ/CLA^*	FEP
K21919	A. baumannii	Sputum	16	4	8	16	4
K22538	A. baumannii	Bile	16	4	16	16	8
K22615	A. baumannii	Sputum	>256	16	32	16	64
K22709	A. baumannii	Sputum	16	4	8	8	4
K22899	A. baumannii	Urine	4	1	2	2	1
K22842	A. baumannii	Sputum	16	4	8	8	4
K23145	A. baumannii	Sputum	16	4	8	8	4
K23308	A. baumannii	Sputum	16	4	8	8	4
K23499	A. baumannii	Sputum	16	4	4	8	2
K18396	A. baumannii	Pus	>256	128	256	>256	64
K18480	A. baumannii	Sputum	>256	128	256	>256	64
K18843	A. baumannii	Bile	256	32	128	256	128
K19583	A. baumannii	Sputum	>256	128	>256	>256	64
K25430	P. aeruginosa	Sputum	16	8	8	16	32
K24686	P. aeruginosa	Urine	>256	256	32	64	>256
K19204	P. aeruginosa	Pus	>256	256	32	64	>256

^* clavulanic acid at a fixed concentration of 4 μg/mL.

Table 4.

Characteristics of SHV-12 or PER-1-producing A. baumannii and P. aeruginosa isolates

Genotype	Isolate	Species	Specimen	Underlying disease	Prognosis	MICs (μg/mL)
Genotype	Isolate	Species	Specimen	Underlying disease	Prognosis	PIP	TZP	CAZ	CAZ/CLA^*	FEP
bla_SHV-12	K19204	P. aeruginosa	Pus	Rectal cancer	Improved	>256	256	32	64	>256
bla_PER-1	KB2660	A. baumannii	Blood	Bronchogenic cancer	Improved	16	4	4	8	4
	KB2919	A. baumannii	Blood	Stomach cancer	Expired	16	4	4	8	4
	K20936	A. baumannii	Sputum	Pulmonary tuberculosis	Improved	16	8	8	16	4
	K21866	A. baumannii	Sputum	Stomach cancer	Improved	16	4	8	16	8
	K23417	A. baumannii	Sputum	Epidural hemorrhage	Improved	16	4	8	8	4
	K20039	A. baumannii	Sputum	Paroxysmal supraventricular tachycardia	Improved	>256	128	>256	>256	>256

^* clavulanic acid at a fixed concentration of 4 μg/mL.

Abbreviations: PIP, piperacillin; TZP, piperacillin-tazobactam; CAZ, ceftazidime; CAZ/CLA, ceftazidime-clavulanic acid; FEP, cefepime.

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