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Characterization of Extended Spectrum $\beta$-Lactamase Genotype TEM, SHV, and CTX-M Producing Klebsiella pneumoniae Isolated from Clinical Specimens in Korea

  • Kim Yun-Tae;Kim Tae-Un;Baik Hyung-Suk
    • Journal of Microbiology and Biotechnology
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    • v.16 no.6
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    • pp.889-895
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    • 2006
  • To investigate the antibiotic-resistant patterns and the gene types of extended-spectrum $\beta$-lactamase (ESBL)-producing Klebsiella pneumoniae, we collected 226 Klebsiella pneumoniae strains from three general hospitals with more than 500 beds in Busan, Korea from September 2004 to October 2005, The minimum inhibitory concentration (MIC) of antibiotics was measured using the Gram-negative susceptibility (GNS) cards of Vitek (Vitek system, Hazelwood Inc., MO, U.S.A.). Of the 226 K, pneumoniae isolates, 65 ESBL-producing K. pneumoniae strains were detected by the Vitek system and confirmed by the double-disk synergy test. TEM (Temoniera) type, SHV (sulfhydryl variable) type, and CTX-M (cefotaxime) type genes were detected by polymerase chain reaction. All 65 K. pneumoniae strains were resistant to ampicillin, cefazolin, cefepime, ceftriaxone, and aztreonam, and 83.0% of the organisms were resistant to ampicillin/sulbactam, 66.1% to tobramycin, 67.6% to piperacillin/tazobactam, 61.5% to ciprofloxacin, and 47.6% to trimethoprim/sulfamethoxazole, and 43.0% to gentamicin. TEM-type ESBLs (TEM-1 type, -52 type) were found in 64.6% (42 of 65) of the isolates, SHV-type ESBLs (SHV-2a type, -12 type, -28 type) in 70.7% (46 of 65) of isolates, and CTX-M-type ESBLS (CTX-M-15 type) in 45% (29 of 65) of isolates. Of the 65 ESBL-producing K. pneumoniae strains, two strains were found to harbor blaSHV-28, which were detected in Korea for the first time. Therefore, more investigation and research on SHV-28 are needed in order to prevent the ESBL type-producing K. pneumoniae from spreading resistance to oxyimino cephalosporin antibiotics.

Random Amplified Polymorphic DNA (RAPD) Analysis for Extended Spectrum ${\beta}-Lactamase$ Producing Klebsiella pneumonia Isolated from Clinical Specimens in Korea

  • Kim Yun-Tae
    • Biomedical Science Letters
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    • v.12 no.3
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    • pp.267-274
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    • 2006
  • Klebsiella pneumoniae is the leading cause of nasocomial infection and the most commonly isolated from clinical specimens. $Extended-spectrum-{\beta}-lactamase$ (ESBL) producing K. pneumoniae infection was associated with a significantly longer duration of hospital stay and greater hospital charges. The purpose of this study is to investigate the antibiotic resistant patterns and the DNA fingerprint types of extended-spectrum ${\beta}-lactamase$ (ESBL) producing K. pneumoniae. 223K. pneumoniae strains were collected from three general hospitals with more than 500 beds in Busan, Korea from September 2004 to October 2005. The minimum inhibitory concentration (MIC) of antibiotics was measured using the Gram negative susceptibility (GNS) cards of VITEK (Vitek system, Hazelwood Inc., MO). Random amplified polymorphic DNA method was used to detect DNA fingerprint of the organisms. Of the 226 K. pneumoniae isolates 65 ESBL-producing K. pneumoniae strains were detected by the Vitek system and confirmed by the double-disk synergy test. All the 65K. pneumoniae strains were resistant cefazolin, cefepime, ceftriaxone and aztreonam, and 83.0% of the organisms were resistant to ampicillin/sulbactam, 66.1% to tobramycin, 67.6% to piperacillin/tazobactam, 61.5% to ciprofloxacin, and 47.6% to trimethoprim/sulfamethoxazole and 43.0% to gentamicin. The RAPD patterns were distincted as 10 types by three random 10-mer primers (208, 272, 277). Among ten type patterns, three types (Ic, IIb, IIIe) were remarkably represented at patient of internal department, nerve surgery department, general surgery department, and neonatal room. These results indicate that RAPD can be useful for DNA of strains typing in the epidemiological investigations. Therefore more investigation are needed in order to prevent the ESBL type-producing K. pneumoniae from spreading resistance to oxyimino cephalosphorin antibiotics.

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Characterization of Extended-Spectrum-$\beta$-Lactamase Genotype TEM, SHV and CTX-M from Clinical Isolates of Klebsiella pneumoniae and Comparison with Antibiotic Susceptibility Test

  • Kim Yun-Tae;Oh Kwang-Seok;Choi Seok-Cheol;Kim Tae-Un
    • Biomedical Science Letters
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    • v.11 no.3
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    • pp.389-396
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    • 2005
  • Resent studies have reported increased isolation of extended-spectrum $\beta-lactamase$ (ESBL) producing strains at several hospital in Korea. We studied to investigate the isolation rates of ESBL strains from clinical isolates of Klebsiella pneumoniae and to characterize differences in types using analyses of genotyping and antibiotic susceptibility test. Antibiotic susceptibility test with confirmation of ESBL by double disk synergy test was performed on the 54 ESBL strains of Klebsiella pneumoniae from a hospital in Busan. Transfer of resistant gene in ESBL strains resistant to 3rd generated antibiotics was confirmed by transconjugation test using E. coli $RG176^{nal(r)}$. blaTEM, blaSHV, blaCTX-M genes were detected by PCR. ESBL producing strains had 100% of resistant rate to ampicillin, azteronam, cefazolin, cefepime and ceftriaxone ($\beta-lactam$ antibiotics). Forty strains of bla TEM$(74\%)$, 41 strains of bla SHV $(76\%)$, 23 strains of bla CTX-M $(43\%)$ were found, respectively. The strains had one or more genes. They had high resistant rates to $\beta-lactam$ antibiotics including cephalosporin. The resistant rates of strains with multiple resistant genes were higher than those of strains with single resistant gene.

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Antimicrobial Resistance of Escherichia coli Isolates from Mussel Mytilus galloprovincialis Farms and Inland Pollution Sources in the Changseon Area, Korea (창선 해역의 지중해담치(Mytilus galloprovincialis) 양식장 및 육상오염원에서 분리한 대장균(Escherichia coli)의 항생제 내성)

  • Kwon, Soon Jae;Lee, Ka Jeong;Jung, Yeoun Joong;Park, Sang Gi;Go, Kyeong Ri;Yang, Ji Hye;Mok, Jong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.5
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    • pp.564-572
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    • 2016
  • In this study, we isolated and characterized Escherichia coli from mussels and inland pollution sources in or in proximity to the Changseon area on the southern coast of Korea in 2014. A total of 147 strains of E. coli were isolated from 54 mussels and 32 pollution-source samples. The susceptibility of the isolates to 24 antimicrobial agents was analyzed. The resistance of E. coli isolates to rifampin was highest at 100%, followed by cephalothin (98.6%), tetracycline (91.8%), amikacin (81.0%), ampicillin (79.6%), cefazolin (79.6%), streptomycin (73.5%), piperacillin (70.7%), gentamicin (37.4%), cefoxitin (35.4%), cefamandole (34.7%), tobramycin (29.9%), amoxicillin/clavulanic acid (24.5%), nalidixic acid (21.8%), trimethoprim (19.0%), chloramphenicol (17.7%), cefotaxime (12.9%), trimethoprim (10.9%), ceftazidime (10.2%), aztreonam (7.5%), imipenem (2.7%), cefepime (2.0%), and cefotetan (0.0%). In addition, the antimicrobial resistance of E. coli isolates from inland pollution sources was slightly greater than or similar to that of isolates from mussels.

Antimicrobial Resistance and Minimum Inhibitory Concentrations of Vibrio parahaemolyticus Strains Isolated from Gomso Bay, Korea (곰소만 해역 해수에서 분리한 장염비브리오(Vibrio parahaemolyticus)의 항균제 내성 및 최소발육억제농도의 구명)

  • Kim, Tae-Ok;Um, In-Seon;Kim, Hee-Dai;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.5
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    • pp.582-588
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    • 2016
  • Seventy-nine Vibrio parahaemolyticus isolates from surface seawater from Gomso Bay, west coast of Korea, were analyzed for the presence of virulence genes and their susceptibility to 30 different antimicrobials. All 79 isolates were examined for the presence of two virulence genes (tdh or trh) using polymerase chain reaction (PCR); however, no isolates possessed either the tdh or trh gene. According to a disk diffusion susceptibility test, all of the strains studied were resistant to oxacillin, penicillin, and vancomycin, followed by ticarcillin (97.5%), ampicillin (96.2%), clindamycin (86.1%), erythromycin (10.1%), streptomycin (7.6%), cefoxitin (6.3%), amikacin (2.5%), and cephalothin (2.5%). However, all of the strains were susceptible to 19 other antimicrobials including cefepime, cefotaxime, chloramphenicol, gentamycin, nalidixic acid, sulfamethoxazole/trimethoprim, and trimethoprim. All 79 isolates (100%) were resistant to four or more classes of antimicrobials, and two strains exhibited resistance to eight antimicrobial agents. The average minimum inhibitory concentrations (MICs) for V. parahaemolyticus for ampicillin, penicillin, ticarcillin, and vacomycin were 946.5, 1,305.9, 1,032.3, and 45.0 µg/mL, respectively.

Uropathogenic Escherichia coli ST131 in urinary tract infections in children

  • Yun, Ki Wook;Lee, Mi-Kyung;Kim, Wonyong;Lim, In Seok
    • Clinical and Experimental Pediatrics
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    • v.60 no.7
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    • pp.221-226
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    • 2017
  • Purpose: Escherichia coli sequence type (ST) 131, a multidrug-resistant clone causing extraintestinal infections, has rapidly become prevalent worldwide. However, the epidemiological and clinical features of pediatric infections are poorly understood. We aimed to explore the characteristics of ST131 Escherichia coli isolated from Korean children with urinary tract infections. Methods: We examined 114 uropathogenic E. coli (UPEC) isolates from children hospitalized at Chung-Ang University Hospital between 2011 and 2014. Bacterial strains were classified into STs by partial sequencing of seven housekeeping genes (adk, fumC, gyrB, icd, mdh, purA, and recA). Clinical characteristics and antimicrobial susceptibility were compared between ST131 and non-ST131 UPEC isolates. Results: Sixteen UPEC isolates (14.0%) were extended-spectrum ${\beta}-lactamase$ (ESBL)-producers; 50.0% of ESBL-producers were ST131 isolates. Of all the isolates tested, 13.2% (15 of 114) were classified as ST131. There were no statistically significant associations between ST131 and age, sex, or clinical characteristics, including fever, white blood cell counts in urine and serum, C-reactive protein, radiologic abnormalities, and clinical outcome. However, ST131 isolates showed significantly lower rates of susceptibility to cefazolin (26.7%), cefotaxime (40.0%), cefepime (40.0%), and ciprofloxacin (53.3%) than non-ST131 isolates (65.7%, 91.9%, 92.9%, and 87.9%, respectively; P<0.001 for all). ESBL was more frequently produced in ST131 (53.3%) than in non-ST131 (8.1%) isolates (P<0.01). Conclusion: ST131 E. coli isolates were prevalent uropathogens in children at a single medical center in Korea between 2011 and 2014. Although ST131 isolates showed higher rates of antimicrobial resistance, clinical presentation and outcomes of patients were similar to those of patients infected with non-ST131 isolates.

Antimicrobial Resistance and Minimum Inhibitory Concentrations of Vibrio parahaemolyticus Strains Isolated from Seawater and Commercial Fisheries (해수 및 시판 수산물에서 분리한 장염비브리오균(Vibrio parahaemolyticus)의 항균제 내성 및 최소발육억제농도의 규명)

  • Cho, Eui-Dong;Kim, Hee-Dai;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.52 no.6
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    • pp.587-595
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    • 2019
  • Eighty-three Vibrio parahaemolyticus isolates from surface seawater in Gomso Bay on the west coast of Korea, and commercial fisheries from Gunsan fisheries center were analyzed for the presence of virulence genes and susceptibility to 30 different antimicrobials. All 83 isolates were examined for the presence of two virulence genes (tdh or trh) using polymerase chain reaction; however, neither gene was found in any of the isolates. A disk diffusion susceptibility test, showed that all of the strains studied were resistant to clindamycin, oxacillin, ticarcillin, and vancomycin, and also revealed varying levels of resistance to ampicillin (98.8%), penicillin G (95.2%), streptomycin (20.5%), cefoxitin (14.5%), amikacin (6.0%), cephalothin (4.8%), and erythromycin (3.6%). However, all of the strains were susceptible to 19 other antimicrobial agents, including cefepime, cefotaxime, chloramphenicol, gentamycin, nalidixic acid, sulfamethoxazole/trimethoprim, and trimethoprim. All 83 isolates (100%) were resistant to five or more classes of antimicrobials, and two strains exhibited resistance to ten antimicrobial agents. The average minimum inhibitory concentrations against V. parahaemolyticus of clindamycin, oxacillin, ticarcillin, and vancomycin were 55.9, 98.3, 499.3, and 44.3 ㎍/mL, respectively. These results provide new insight into the necessity for seawater sanitation in Gomso Bay and commercial fisheries, and provide evidence to help reduce the risk of contamination by antimicrobial-resistant bacteria.

L-glutamine:D-fructose-6-phosphate Aminotransferase as a Key Protein Linked to Multidrug Resistance in E. coli KD43162

  • Lee, Sung-Eun;Jung, Tae-Jeon;Park, Byeoung-Soo;Kim, Byung-Woo;Lee, Eun-Woo;Kim, Hye Jin;Yum, Jong Hwa
    • Journal of Applied Biological Chemistry
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    • v.58 no.3
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    • pp.227-232
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    • 2015
  • A microarray study has been employed to understand changes of gene expression in E. coli KD43162 resistant to ampicillin, ampicillin-sulbactam, piperacillin, piperacillin-tazobactam, cefazolin, cefepime, aztreonam, imipenem, meropenem, gentamicin, tobramycin, ciprofloxacin, levofloxacin, moxifloxacin, fosfomycin, and trimethoprim-sulfamethoxazole except for amikacin using disk diffusion assay. Using Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and MALDI-TOF MS analyses, 36 kDa of outer membrane proteins (OMPs) was found to be deleted in the multidrug resistant E. coli KD 43162. Microarray analysis was used to determine up- and down-regulated genes in relation to multidrug resistant E. coli KD43162. Among the up-regulated genes, these genes were corresponded to express the proteins as penicillin-binding proteins (PBPs), tartronate semialdehyde reductase, ethanolamine utilization protein, shikimate kinase I, allantoinase, predicted SAM-dependent methyltransferase, L-glutamine: D-fructose-6-phosphate aminotransferase (GFAT), phospho-glucosamine mutase, predicted N-acetylmannosamine kinase, and predicted N-acetylmannosamine-6-P epimerase. Up-regulation of PBPs, one of primary target sites of antibiotics, might be responsible for the multidrug resistance in E. coli with increasing amount of target sites. Up-regulation of GFAT enzyme may be related to the up-regulation of PBPs because GFAT produces N-acetylglucosamine, a precursor of peptidoglycans. One of GFAT inhibitors, azaserine, showed a potent inhibition on the growth of E. coli KD43162. In conclusion, up-regulation of PBPs and GFATs with the loss of 36 kDa OMP refers the multidrug resistance in E. coli KD 43162.

A Case of Ochrobactrum anthropi Infection after Using Medicinal Plants (약초 복용 후 Ochrobactrum anthropi에 감염 1예)

  • Cho, Seang-Sig;Cheun, Jai-Woo;Jeun, Chun-Bae;Park, Sang-Muk;Jang, Sook-Jin;Moon, Dae-Soo;Park, Young-Jin
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.1
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    • pp.22-25
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    • 2006
  • Ochrobactrum anthropi, previously known as Achromobacter species biotypes 1 and 2 (CDC groups Vd-1, Vd-2), belong to the groups of non-Enterobacteriaceae- nonfermentative Gram negative bacilli. Achromobacter is not presently a recognized genus. Achromobacter xylosoxidans has been transferred to genus Alcaligenes as A. xylosoxidans subsp. xylosoxidans, and "Achromobacter" sp. group Vd has been named Ochrobactrum anthropi. O. anthropi was isolated from a blood culture. Organisms were identified as O. anthropi by use of the biochemical test and the VITEK 2(bioMerieux, USA). The Organism was susceptible only to colistin, imipenem, meropenem, and tetracycline, but were resistant to amikacin, aztreonam, cefepime, ceftazidime, cefpirome, ciprofloxacin, gentamicin, isepamcin, netilmicin, pefloxacin, piperacillin, piperacillin/tazobactam, ticarcillin, ticarcillin/clavulanic acid, tobramycin, and trimethoprim/sulfamethoxazole. We report the clinical and microbiologic characteristics of O. anthropi infection in the patient. This is the first case of O. anthropi infection after using a plant as medicine at Chosun University Hospital.

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Prevalence of antimicrobial resistance and integrons in extended-spectrum β-lactamases producing Escherichia coli isolated from Nakdong and Gumho river (낙동강과 금호강에서 분리된 광범위 베타 락탐 분해효소 생성 Escherichia coli 내 항균제 내성 및 integrons의 분포)

  • Cho, Jae-Keun;Kim, Hwan-Deuk;Kwon, Soon-Hyo;Kim, Jin-Hyun;Jang, Sung-Il;Park, Choi-Kyu;Kim, Ki-Seuk
    • Korean Journal of Veterinary Service
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    • v.37 no.1
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    • pp.19-27
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    • 2014
  • This study was conducted to investigate the antimicrobial resistance, presence of ${\beta}$-lactamase genes and integrons in 83 ESBL-producing Escherichia coli isolated from Nakdong river and Geumho river in Daegu. Among the ${\beta}$-lactam antimicrobials, all isolates were resistant to ampicillin, cephalothin, cefamandole and cefotaxime, followed by piperacillin (98.8%), ampicillin/sulbactam (86.7%), aztreonam (60.2%) and cefepime (59.0%), whereas resistance to piperacillin/tazobacram, ticarcillin/clavulanic acid and cefoxitin was less than 30%. Many of the ESBL-producing Escherichia coli were also resistant to non-${\beta}$-lactams antimicrobials such as nalidixic acid (83.1%), sulfonamides (72.3%), ciprofloxacin (62.7%) and gentamicin (38.6%). All isolates showed resistance to seven or more antimicrobial agents. The most frequently detected gene was $bla_{TEM+CTX-M}$ (49.4%), followed by $bla_{CTX-M}$ (27.7%), $bla_{TEM}$ (6.0%) and $bla_{OXA}$ (1.2%). But $bla_{SHV}$ was not found. Class 1 integrons were found in 61.4% (51 isolates) of isolates, however, class 2 and 3 integrons were not detected. The results showed water from Nakdong river and Geumho river is contaminated with ESBL-producing E. coli isolates. These results suggest the need for further investigation of antibiotic resistant bacteria to prevent public health impacts in the water environment.