Annals of Laboratory Medicine

Korean Society for Laboratory Medicine (대한진단검사의학회)
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Prevalence of Class A Extended-Spectrum ${\beta}$-Lactamases in Clinical Isolates of Acinetobacter baumannii and Pseudomonas aeruginosa

임상검체에서 분리된 Acinetobacter baumannii와 Pseudomonas aeruginosa의 Class A Extended-Spectrum ${\beta}$-Lactamase 생성 현황

Oh, Se-Jin;Lee, Sang-Uk;Hwang, Hyun-Yong;Bae, Il-Kwon;Jo, Hyun-Soo;Lee, Byung-Ho;Jeong, Seok-Hoon
오세진;이상욱;황현용;배일권;조현수;이병호;정석훈

  • Published : 20060000
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Abstract

Background : Prevalence of class A extended-spectrum ${\beta}$-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 $bla_{TEM}$, $bla_{SHV}$, $bla_{CTX-M}$, $bla_{PER-1}$, $bla_{VEB}$, and $bla_{GES/IBC}$ 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 $bla_{PER-1}$ (n=6), and $bla_{SHV-12}$ 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 ${\beta}$-lactam antibiotics for the treatment of A. baumannii and P. aeruginosa infections.

배경 : 국내에서 분리되는 Escherichia coli나 Klebsiella pneumoniae의 class A extended-spectrum ${\beta}$-lactamase (ESBL) 생성현황에 대해서는 여러차례 조사가 있었지만, Acinetobacter baumannii나 Pseudomonas aeruginosa의 이들 효소 생성 현황에 대한 조사는 매우 드물다. 본 연구에서는 부산의 한 대학병원에서 분리된 A. baumannii와 P. aeruginosa를 대상으로 TEM, SHV, PER, VEB, GES/IBC 등 class A의 ESBL 생성 현황을 조사하며, ESBL의 유전형을 규명고자 하였다. 방법 : 2004년 6-9월에 고신대학교 복음병원 환자의 임상검체에서 분리된 A. baumannii와 P. aeruginosa를 대상으로 하였다. 항균제 감수성은 디스크 확산법과 한천 희석법으로 시험하였고, ESBL 생성은 double-disk synergy 시험으로 확인하였다. Ceftazidime 내성 전달성은 접합으로 시험하였다. Isoelectric focusing으로 ${\beta}$-lactamase의 pI값을 측정하였으며 중합연쇄반응으로 TEM형, SHV형, CTX-M형, PER-1형, VEB형, IBC형 및 GES형 효소의 유전자를 검출하였고, 중합연쇄반응 산물의 염기서열을 양방향으로 분석하였다. 결과 : 시험기간 중 총 58주의 A. baumannii와 77주의 P. aeruginosa가 환자의 임상검체에서 검출되었다. A. baumannii 3주(5.2%)와 P. aeruginosa 4주(5.2%)는 imipenem 디스크와 ceftazidime 디스크 사이에 synergy 현상을 보였으며, A. baumannii 1주(1.7%)와 P. aeruginosa 2주(2.6%)는 cefoxitin 디스크와 ceftazidime 디스크 사이에 synergy 현상을 보였다. 대상균주 중 A. baumannii 6주는 $bla_{PER-1}$ 유전자를 지니고 있었으며, $bla_{SHV-12}$ 유전자를 지닌 P. aeruginosa 1주 및 $bla_{TEM-116}$ 유전자를 지닌 A. baumannii 13주와 P. aeruginosa 3주도 검출되었다. 결론 : 이상의 결과에서 부산의 한 대학병원에서 분리되는 A. baumannii 중 가장 흔한 class A ESBL은 PER-1이며, SHV-12를 생성하는 P. aeruginosa의 존재도 증명하였다. 이들 ESBL 생성 A. baumannii와 P. aeruginosa 균주의 확산은 향후 광범위 ${\beta}$-lactam 항균제 임상적 효용성을 제한할 것으로 우려된다.

Keywords

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