Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Characterization of Plasmid-Mediated SHV-11 β-lactamase Gene of Klebsiella pneumoniae Isolated from the Clinical Specimens

임상검체로부터 분리한 플라스미드 매개성 SHV-11 β-lactamase 유전자의 특성

  • Kim, Yun-Tae (Department of Microbiology Genetics, Seoul Medical Science Institute/Seoul Clinical Laboratories) ;
  • Lee, Sang-Hoo (Department of Microbiology Genetics, Seoul Medical Science Institute/Seoul Clinical Laboratories) ;
  • Jang, Ji-Hyun (Department of Microbiology Genetics, Seoul Medical Science Institute/Seoul Clinical Laboratories) ;
  • Kim, Tae-Un (Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan) ;
  • Choi, Seok-Cheol (Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan) ;
  • Baik, Hyung-Suk (Division of Biological Science, Pusan National University) ;
  • Lee, Kyoung-Ryul (Department of Microbiology Genetics, Seoul Medical Science Institute/Seoul Clinical Laboratories) ;
  • Yoon, Hye-Ryoung (Department of Microbiology Genetics, Seoul Medical Science Institute/Seoul Clinical Laboratories) ;
  • Kim, Young-Jin (Department of Microbiology Genetics, Seoul Medical Science Institute/Seoul Clinical Laboratories)
  • Published : 2009.12.30
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Abstract

In this study, we characterized extended-spectrum $\beta$-lactamase (ESBL)-producing Enterobacteriaceae isolated from clinical specimens in Korea and found two strains harboring plasmid-mediated $bla_{SHV-11}$, Klebsiella pneumoniae. First, the isolates were detected using the Vitek system and confirmed by the double-disk synergy test. The classification of gene coding for ESBL was also performed by polymerase chain reactions and followed by DNA sequencing. The transmission of genes was confirmed by transconjugation and transformation. Resistant expression of transformants was determined by broth microdilution minimal inhibitory concentration test. Genotypic analysis revealed that one strain harbored the $bla_{TEM-1}$, $bla_{SHV-11}$ and $bla_{CTX-M-15}$ and the other strain harbored the $bla_{SHV-11}$ and $bla_{CTX-M-15}$. They showed high resistance to oxyiminocephalosphorins (3rd-generation cephalosporins), while the transformant containing only $bla_{SHV-11}$ did not show any resistance to the antibiotics.

Chromosomal 인 SHV-11 $\beta$-lactamase가 plasmid를 매개로 다른 균주로 전달 되는 현상은 흔하지 않다. 본 연구에서는 플라스미드성 SHV-11 $\beta$-lactamase를 동시에 가지고 있는 ESBL생성 두 균주를 검출하였다. 따라서 이들 균주에 대한 유전적 특성과 임상적 의의에 대해 알아보고자 하였다. Vitek system과 이중디스크확산법을 이용하여 ESBL생성균주를 검출하였고, PCR과 DNA 염기서열분석을 이용하여 SHV-11 $\beta$-lactamase를 가지고 있는 ESBL생성균주를 확인 하였다. 이들 균주를 교차접합실험과 형질전환실험을 이용하여 유전자전이를 확인하고 액체배지 희석법으로 3세대 cephalosphorin 항생제에 대한 최소억제농도를 측정하였다. 이들 균주의 유전형 분석결과는 SHV-11 $\beta$-lactamase 유전자와 CTX-M-15 ESBL 유전자를 동시에 가지고 있었다. 3세대 cephalosphorin 항생제에 대한 최소억제농도는 SHV-11 $\beta$-lactamase와 CTX-M-15 ESBL 유전자를 동시에 가지고 있는 균주에서 $64{\mu}g/ml$ 이상이었고, SHV-11 $\beta$-lactamase 만을 가지고 재조합 한 균주에서 $0.5{\mu}g/ml$ 이하로 나타났다.

Keywords

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