• Title/Summary/Keyword: Beta-lactamases

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Characterization of Noble AmpC-Type $\beta$-Lactamases Among Clinical Isolates Using New Expression/Secretion Vector (발현ㆍ분비 벡터 및 임상 균주가 생성하는 신규 AmpC-type $\beta$-lactamase의 특성)

  • 정하일;성광훈;이정훈;장선주;이상희
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.104-110
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    • 2004
  • To determine evolution and genotype of new chromosomal AmpC $\beta$-lactamases among clinical isolates of Enterobacter species, we performed antibiotic susceptibility testing, pI determination, sequencing, and phy-logenetic analysis using developed expression/secretion vector. Six isolates have shown to produce AmpC $\beta$-lactamases. Six genes of AmpC $\beta$-lactamases that are responsible for the resistance to cephamycins (cefoxitin and cefotetan), amoxicillin, cephalothin, and amoxicillin-clavulanic acid were cloned and characterized in pMSG12119. Insert fragment containing the ampC genes was sequenced and found to have an open reading frame coding for 381-amino-acid $\beta$-lactamase. The nucleotide sequence of four ampC genes ($bla_EcloK992004.l$, $bla_EcloK995120.1$, $bla_EcloK99230$, and $bla_EareK9911729$) shared considerable homology with that of chromosomal ampC gene ($bla_EcloMHN1$) of E. cloacae MHN1 (more than 99.6% identity). The sequences of two ampC genes ($bla_EcloK9973$ and $bla_EcloK9914325$) showed close similarity to the chromosomal ampC gene ($bla_EcloQ908R$) of E. clo-acae 908R (99.7% identity). The results from phylogenetic analysis suggested that six ampC genes could be originated from $bla_EcloMHN1$ / or $bla_EcloQ908R$ / MIC patterns and exact pI values of six transformants indicated that the developed expression/secretion vector (pMSG1219) was suitable for the characterization of foreign genes in E. coli strain.

Definitive Nomenclature of GES/IBC-Type Extended-Spectrum ${\beta}-Lactamases$

  • Weldhagen Gerhard F.;Kim, Bok-Hee;Cho, Chan-Hwi;Lee, Sang-Hee
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1837-1840
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    • 2006
  • Because there are no unified nomenclature systems for either GES-type or IBC-type extended-spectrum ${\beta}-lactamases$ (ESBLs), we propose a unified and definitive nomenclature system for GES/IBC-type ESBLs. This proposed nomenclature update is greatly helpful in two points: (i) it would not confuse microbiologists studying GES-type ESBLs, fundamentally preventing misleading nomenclature of these antibiotic resistance genes, and (ii) the definitive renaming of GES/IBC-type ESBLs can help some researchers to correctly designate new GES-type ESBLs such as novel enzymes identified trom some nationwide surveys.

Characterization of a New ${\beta}$-Lactamase Gene from Isolates of Vibrio spp. in Korea

  • Jun, Lyu-Jin;Kim, Jae-Hoon;Jin, Ji-Woong;Jeong, Hyun-Do
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.555-562
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    • 2012
  • PCR was performed to analyze the ${\beta}$-lactamase genes carried by ampicillin-resistant Vibrio spp. strains isolated from marine environments in Korea between 2006 and 2009. All 36 strains tested showed negative results in PCR with the primers designed from the nucleotide sequences of various known ${\beta}$-lactamase genes. This prompted us to screen new ${\beta}$-lactamase genes. A novel ${\beta}$-lactamase gene was cloned from Vibrio alginolyticus KV3 isolated from the aquaculture water of Geoje Island of Korea. The determined nucleotide sequence (VAK-3 ${\beta}$-lactamase) revealed an open reading frame (ORF) of 852 bp, encoding a protein of 283 amino acids (aa), which displayed low homology to any other ${\beta}$-lactamase genes reported in public databases. The deduced 283 aa sequence of VAK-3, consisting of a 19 aa signal peptide and a 264 aa mature protein, contained highly conserved peptide segments specific to class A ${\beta}$-lactamases including the specific amino acid residues STFK (62-65), SDN (122-124), E (158), and RTG (226-228). Results from PCR performed with primers specific to the VAK-3 ${\beta}$-lactamase gene identified 3 of the 36 isolated strains as V. alginolyticus, Vibrio cholerae, and Photobacterium damselae subsp. damselae, indicating the utilization of various ${\beta}$-lactamase genes including unidentified ones in ampicillin-resistant Vibrio spp. strains from the marine environment. In a mating experiment, none of the isolates transfered the VAK-3 ${\beta}$-lactamase gene to the Escherichia coli recipient. This lack of mobility, and the presence of a chromosomal acyl-CoA flanking sequence upstream of the VAK-3 ${\beta}$-lactamase gene, led to the assumption that the location of this new ${\beta}$-lactamase gene was in the chromosome, rather than the mobile plasmid. Antibiotic susceptibility of VAK-3 ${\beta}$-lactamase was indicated by elevated levels of resistance to penicillins, but not to cephalosporins in the wild type and E. coli harboring recombinant plasmid pKV-3, compared with those of the host strain alone. Phylogenetic analysis showed that VAK-3 ${\beta}$-lactamase is a new and separate member of class A ${\beta}$-lactamases.

Multiplex PCR for Detection of Quinolone Resistance qnr Genes in Extended-Spectrum β-Lactamase Producing Escherichia coli and Klebsiella pneumoniae (Multiplex PCR을 이용한 Extended-Spectrum β-Lactamase 생성 Escherichia coli와 Klebsiella pneumoniae의 Quinolone 내성 qnr유전자 검출)

  • Yang, Byoung-Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.3
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    • pp.161-166
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    • 2007
  • To develop a rapid and reliable single-tube-based PCR technique for detection simultaneously the quinolone resistance qnrA, qnrB and qnrS genes. After multiple alignment, primers were designed to detect known qnr variants. I was used for A total of 43 extented-spectrum ${\beta}$-lactamases (ESBLs) producing Escherichia coli and Klebsiella pneumoniae isolated from university hospital were tested for screening, as with qnr genes. In optimized conditions, all positive controls confirmed the specificity of the PCR primers. Out of 43 isolates, qnrA genes were detected 19 (44.2%), qnrB genes 5 (11.7%), qnrS genes 15 (34.9%) and 8 (18.6%) isolates were not detected. I report here a fast and reliable technique for rapid screening of qnr positive strains to be used for epidemiological surveys.

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Typing of Extended-Spectrum ${\beta}-Lactamase$ of Escherichia coli and Klebsiella pneumoniae Isolated from Rivers in Busan, Korea (부산지역 하천에서 분리된 장내세균 Escherichia coli와 Klebsiella pneumoniae의 광범위 베타 락탐 분해효소 (Extended-Spectrum ${\beta}-Lactamase$)에 대한 유형별 분류)

  • Lee, Hun-Ku;Kim, Hye-Jin;Kim, Gun-Do
    • Korean Journal of Microbiology
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    • v.43 no.2
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    • pp.116-123
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    • 2007
  • The purpose of this study was typing the plasmid mediated extended spectrum ${\beta}-lactamases$ produced by enteric bacteria isolated from rivers in Pusan. Six strains of Eschericha coli and fifteen strains of Klebsiella pneumoniae transferred their plasmid mediated extended spectrum ${\beta}-lactamase$ genes to the recipient strain Eschericha coli J53 $Azid^{R}$. The plasmid mediated extended spectrum ${\beta}-lactamase$ genes were sequenced directly after PCR and the types were determined by the BCM Search Launcher and GenBank nucleotid database. Determined types of the plasmid mediated extended spectrum ${\beta}-lactamases$ were TEM-52 and SHV-12. TEM-52 was isolated from both Escherichia coli and Klebsiella pneumoniae. However SHV-12 was isolated from Klebsiella pneumoniae only. The results indicated that the plasmid mediated extended spectrum ${\beta}-lactamase$ producing bacteria spreded over the area of clinical to the nature in Korea.

Detection Rate of Extended-Spectrum ${\beta}$-Lactamase Producers in Klebsiella pneumoniae and Escherichia coli Isolated at Yeungnam University Medical Center (영남대학교 의과대학 부속병원에서 동정된 Klebsiella pneumoniae와 Escherichia coli의 Extended-Spectrum ${\beta}$-Lactamase생성 빈도)

  • Lee, Chae-Hoon;Lee, Ho-Chan;Kim, Kyung-Dong;Lee, Tae-Su
    • Journal of Yeungnam Medical Science
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    • v.16 no.2
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    • pp.270-276
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    • 1999
  • Background: Extended-spectrum ${\beta}$-lactamases(ESBL) are enzymes that confer resistance to oxyimino-${\beta}$-lactams as well as to penicillins and cephalosporins. Strains of Klebsiella pneumoniae and Escherichia coli that produce ESBL have been increasingly prevalent in many countries. The purpose of this study was to investigate the ESBL production rate of K. pneumoniae and E. coli at the in Yeungnam University Medical Center. Materials and Methods: Thirty-one isolates of K pneumoniae and twenty-five isolates of E. coli were examined for ESBL by double disk synergy test, using 20/$10{\mu}g$ ticarcillin/clavulanic acid and $30{\mu}g$ oxyimino-${\beta}$-lactam(ceftazidime, ceftaxime, ceftriaxone and aztreonam) disks. Results: Fifty-two percent of K. pneumoniae and sixteen percent of E. coli isolates revealed double disk synergism. Majority of ESBL-producing strains(fifty-five percent) were isolated from patients in the intensive care unit. Conclusion: ESBL production of K. pneumoniae and E. coli were also common at the Yeungnam University Medical Center and pose a serious problem for antimicrobial therapy.

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Synthesis and Antibacterial Activities of Triphenyltin Cephalosporins

  • Park, Sang-Woo;Kim, You-Seung;Chung, Young-Keun
    • Archives of Pharmacal Research
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    • v.12 no.3
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    • pp.231-232
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    • 1989
  • Although it has been known that organometallic $\beta$-lactam compounds improve the resistance to $\beta$-lactamases as well as their pharmacological activities, only a few results on organometallic $\beta$-lactam antibiotics were reported. In the course of our extensive study on the development of new cephalosporins, we were interested in organotin compounds since they show some biological activities.

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Synthetic Cephalosporin Derivatives

  • Oh, Chang-Hyun;Park, Sang-Woo;Cho, Jung-Hyuck
    • Bulletin of the Korean Chemical Society
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    • v.11 no.4
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    • pp.323-327
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    • 1990
  • The synthesis and some biological properties of $7{\beta} $-[2-(Z)-(2-aminothiazole-4-yl)-2-(N-substitutedcar bonyl)ethoxyiminoacetamido]-3-vinyl-3-cephem-4- carboxylic acid are described. The effect of substituents on the carbamoly group in the 7-side chain were investigated in order to improve antibacterial activities. Two of these new orally active $7{\beta} $-lactam derivatives showed wide expanded antimicrobial activities against Gram-positive and Gram-negative bacteria, including Pseudomonas aeruginosa, as well as good stability to $7{\beta} $ -lactamases.

Selection of Clinically Isolated Strains for Evaluation of the Newly Synthesized Antibiotics (새로운 $\beta$-lactam계 항생물질 개발을 위한 검정용 균주의 개발)

  • 김대진;최금화;김숙경;최성숙;김병각;강창율;최응칠
    • YAKHAK HOEJI
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    • v.39 no.2
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    • pp.131-136
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    • 1995
  • Clinically isolated bacterial strains resistant to almost of all the clinically superior .betha.-lactam antibiotics can be used to screen the promising ones among the newly synthesized $\beta$-lactam antibiotics. To select the resistant strains, the susceptibility of 389 strains of S. aureus, 144 strains of coagulase negative staphylococci, 509 strains of E. coli, 115 strains of E. cloacae and 187 strains of P. aeruginosa to methicillin, ampicillin, piperacillin and gentamicin was determined. The susceptibility of 19 bacterial strains selected through the first screening to cefixime, cefotiam, cefotaxime, flomoxef, cepfirome, cefdnir, SCE-2787, panipenem and imipenem was determined. Four strains of S. aureus finally selected have high degree of resistance to almost of all $\beta$-lactam antibiotics used and also produce $\beta$-lactamases. These 4 strains of S. aurues can be used to screen effectively the promising $\beta$-lactam antibiotics among the numerous numbers of the newly synthesized $\beta$-lactam antibiotics.

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