• Title/Summary/Keyword: Clinical isolates

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Detection of Extended-Spectrum β-Lactamase Producing Klebsiella pneumoniae by Multiplex Polymerase Chain Reaction (Multiplex Polymerase Chain Reaction을 이용한 Extended-Spectrum β-Lactamase 생성 Klebsiella pneumoniae 균주의 검출)

  • Yang, Byoung-Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.173-178
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    • 2006
  • The production of extended-spectrum ${\beta}$-lactamases ($ESBL_S$) is the main mechanism of bacterial resistance to third-generation cephalosporins and monobactams, whose prevalence varies depending on the different geographical areas. In the last years it has increased notably to the point of being considered a health problem of great importance. The characterization of the ESBLs producing Klebsiella penumoniae strains present in clinical isolates is time-consuming. I describe here the development of a new system, which consists of a multiplex PCR. I found 51 K. pneumoniae strains to be presumptive strains ESBLs producers by clinical and laboratory standards institute (CLSI) guidelines. The double disc synergy test showed 47 positive K. pneumoniae, which were K. pneumoniae isolates. All ESBLs producing K. pneumoniae strains were resistant to antibiotic amikacin, gentamicin and ciprofloxacin. By multiplex PCR analysis, $bla_{TEM}$ gene in 17 strains 44 $bla_{SHV}$ genes and $bla_{CTX}$ genes in 33 strains were identified. In this study, the multiplex polymerase chain reaction (PCR) assay was a good method to detect and differentiate ESBLs producing K. penumoniae strains in clinical isolates.

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Antibiotic Susceptibility Pattern and Molecular Typing By PCR-RAPD Analysis of Clinical and Environmental Isolates of Pseudomonas aeruginosa

  • Oluborode, O.B.;Smith, S.I.;Seriki, T.A.;Fowora, M.;Ajayi, A.;Coker, A.O.
    • Microbiology and Biotechnology Letters
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    • v.46 no.4
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    • pp.434-437
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    • 2018
  • Pseudomonas aeruginosa accounts for a significant proportion of nosocomial infections. This study examined the antimicrobial susceptibility pattern and clonal relatedness of P. aeruginosa isolates of clinical and environmental origin. These isolates displayed susceptibility to levofloxacin, ciprofloxacin, gentamicin, imipenem, and ceftazidime of 65.0%, 62.5%, 90.0%, 100%, and 85%, respectively. PCR-RAPD analysis of the P. aeruginosa isolates revealed marked variation. No correlation was observed between the antibiotic resistance profiles and the DNA typing patterns.

Genetic analyses of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea

  • Yu, Hak-Sun;Choi, Kyung-Hee;Kim, Hyo-Kyung;Kong, Hyun-Hee;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.161-170
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    • 2001
  • We conducted both the small subunit ribosomal DNA (SSU rDNA) polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and mitochondrial (mt) DNA RFLP analyses for a genetic characterization of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea. Twenty-three strains of Acanthamoeba from the American Type Culture Collection and twelve clinical isolates from Korean patients were used as reference strains. Thirty-nine isolates from contact lens storage cases were classified into seven types (KA/LS1 , KA/LS2, KA/LS4, KA/LS5, KA/LS7 KA/LS18, KA/LS31). Four types (KA/LS1 , KA/LS2, KA/LS5, KA/LS18) including 33 isolates were regarded as A. castellanii complex by riboprints. KA/LS1 type was the most predominant (51.3%) in the present survey area, followed by KA/LS2 (20.9%), and KA/LSS (7.7%) types. Amoebae of KA/LS1 type had the same mtDNA RFLP and riboprint patterns as KA/E2 and KA/E12 strains, clinical isolates from Korean keratitis patients. Amoebae of KA/LS2 type had the identical mtDNA RFLP patterns with A. castellanii Ma strain, a corneal isolate from an American patient as amoebae of KA/LS5 type, with KA/E3 and KA/E8 strains from other Korean keratitis patients. Amoebae of KA/LS 18 type had identical patterns with JAC/E1, an ocular isolate from a Japanese patient. Three types , which remain unidentified at species level, were not corresponded with any clinical isolate in their mtDNA RFLP and riboprint patterns. Out of 39 isolates analyzed in this study, mtDNA RFLP and riboprint patterns of 33 isolates (84.6%) were identical to already known clinical isolates, and therefore, they may be regarded as potentially keratopathogenic. These results suggest that contact lens wearers in Seoul should pay more attention to hygienic maintenance of contact lens storage cases for the prevention of Acanthamoeba keratitis.

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First Detection of $bla_{IMP-1}$ in Clinical Isolate Multiresistant Acinetobacter baumannii from Korea

  • Jeong Seok-Hoon;Bae Il-Kwon;Sohn Seung-Ghyu;Park Kwang-Ok;An Young-Jun;Sung Kwang-Hoon;Jang Seon-Ju;Heo Myong-Jin;Yang Ki-Suk;Lee Sang-Hee
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1377-1383
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    • 2006
  • Among 46 Acinetobacter baumannii isolates collected in 2004, two imipenem-resistant isolates were obtained from clinical specimens taken from patients hospitalized in Busan, Republic of Korea. Two carbapenemase-producing isolates were further investigated to determine the mechanism of resistance. These isolates were analyzed by antibiotic susceptibility testing, microbiological tests of carbapenemase activity, determination of pI, transconjugation test, enterobacterial repetitive consensus (ERIC)-PCR, and DNA sequencing. Two cases of infection by A. baumannii producing the IMP-1 ${\beta}$-lactamase were detected. The isolates were characterized by a modified cloverleaf synergy test and EDTA-disk synergy test. Isoelectric focusing of crude bacterial extracts revealed nitrocefin-positive bands with a pI value of 9.0. PCR amplification and characterization of the amplicons by direct sequencing indicated that the isolates carried a $bla_{IMP-l}$ determinant. The isolates were characterized by a multidrug resistance phenotype, including penicillins, extended-spectrum cephalosporins, carbapenems, and aminoglycosides. These results indicate that the observed imipenem resistance of two Korean A. baumannii isolates was due to the spread of an IMP-1-producing clone. Our microbiological test of carbapenemase activity is simple to screen class B metallo-${\beta}$-lactamase-producing clinical isolates to determine their clinical impact and to prevent further spread. This study shows that the $bla_{IMP-l}$ resistance determinant, which is emerging in Korea, may become an emerging therapeutic problem, since clinicians are advised not to use extended-spectrum cephalosporins, imipenem, and aminoglycosides. This observation emphasizes the importance of having effective control measures in Asian hospitals, such as early detection of colonized patients, isolation procedures, and a judicious use of antibiotics.

Identification of Fusobacterium nucleatum isolated from Korean by F. nucleatum subspecies-specific DNA probes (Dot blot hybridization법을 이용한 Fusobacterium nucleatum 아종-특이 DNA 프로브의 특이성 평가)

  • Kim, Hwa-Sook;Kook, Joong-Ki
    • Journal of Korean society of Dental Hygiene
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    • v.6 no.4
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    • pp.311-324
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    • 2006
  • The purpose of this investigation was to evaluate of the specificity of Fusobacterium nucleatum subspecies-specific DNA probes using dot blot hybridization. To confirm whether the clinical isolates were F. nucleatum or not, 16S rDNA of them were cloned and sequenced. The sequencing data were used in homology search with database of GenBank. When the homology was above 98% compared with the nucleotide sequence of a certain bacteria, it was judged as the same species with the bacteria. 23 strains of F. nucleatum were isolates from subgingival plaque of periodontitis patient. The clinical isolates of F. nucleatum were classified into 10 groups using phylogenetic analysis of 16S rDNA sequence. F. nucleatum subspecies nucleatum-specific DNA probe Fu4(1.3 kb) reacted with genomic DNAs from 8 type strains of F. nucleatum and it reacted strongly with those from 8 clinical isolates. The Fp4(0.8 kb) reacted with F. nucleatum subsp. polymorphum ATCC 10953 and one clinical isolates. Fv35(1.9 kb) and Fs17(8.2 kb) probes reacted with genomic DNAs from F. nucleatum subsp. vincentii ATCC 49256 and F. nucleatum subsp. fusiform ATCC 51190, respectively. Our results showed that it is not enough to evaluate the specificity of F. nucleatum subspecies-specific DNA probes with only dot blot hybridization. Therefore, Southern blot analysis will be necessary to confirm the specificity of F. nucleatum subspecies-specific DNA probes.

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Antibiotic Resistance Patterns of Acinetobacter baumannii and Pseudomonas aeruginosa from a Hospital in Daegu City Area (대구지역병원에서 분리된 Acinetobacter baumannii와 Pseudomonas aeruginosa의 항생제 내성현황)

  • Kim, Su-Jung;Lee, Jae-Sik
    • Korean Journal of Clinical Laboratory Science
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    • v.40 no.2
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    • pp.75-79
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    • 2008
  • 86 clinical isolates of Acinetobacter baumannii and 116 clinical isolates of Pseudomonas aeruginosa strains isolated from clinical specimens were collected from a hospital in Daegu city area. We investigated the Antimicrobial susceptibility patterns of A. baumannii and P. aeruginosa isolated from sputum, urine, wound, blood, nasal swab, body fluid. The antimicrobial resistance of A. baumannii were shown 96% for piperacillin, carbenicillin 82%. cefotaxime 78%, ciprofloxacin 77%, sulfamethoxazole/trimethoprime 76%, ceftazidime 75%, tobramycin 72%. For P. aeruginosa, the resistance of cefotaxime and sulfamethoxazole/trimethoprime were 100%, carbenicillin 49%, piperacillin 47%, ticarcillin 45%, ticarcillin/ clavulanic acid 40%.

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Evaluation of Ciclopirox as a Virulence-modifying Agent Against Multidrug Resistant Pseudomonas aeruginosa Clinical Isolates from Egypt

  • Zakaria, Azza S.;Edward, Eva A.;Mohamed, Nelly M.
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.651-661
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    • 2019
  • Targeting the pathogen viability using drugs is associated with development of drug resistance due to selective pressure. Hence, there is an increased interest in developing agents that target bacterial virulence. In this study, the inhibitory effect of ciclopirox, an antifungal agent with iron chelation potential, on the microbial virulence factors was evaluated in 26 clinical MDR Pseudomonas aeruginosa isolates collected from Alexandria Main University Hospital, a tertiary hospital in Egypt. Treatment with 9 ㎍/ml ciclopirox inhibited the hemolytic activity in 70% isolates, reduced pyocyanin production, decreased protease secretion in 46% isolates, lowered twitching and swarming motility, and decreased biofilm formation by 1.5- to 4.5-fold. The quantitative real-time PCR analysis revealed that treatment with ciclopirox downregulated the expression levels of alkaline protease (aprA) and pyocyanin (phzA1). Ciclopirox is used to treat hematological malignancies and the systemic administration of ciclopirox is reported to have adequate oral absorption with a satisfactory drug safety profile. It is important to calculate the appropriate clinical dose and therapeutic index to reposition ciclopirox from a topical antifungal agent to a promising virulence-modifying agent agent against P. aeruginosa, a problematic Gram-negative pathogen.

Isolation Rate of Campylobacter fetus subsp. jejuni from Enteritis Patients (장염환자에서의 Campylobacter fetus subsp. jejuni 분리율)

  • Chong, Yun-Sop;Yi, Kui-Nyung;Lee, Sam-Uel Y.
    • The Journal of the Korean Society for Microbiology
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    • v.17 no.1
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    • pp.43-47
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    • 1982
  • C. fetus suhsp. jejuni has been reported to be an important enteric pathogen in many parts of the world. Although the infection has been reported in Korea, the incidence is not known. In this study the results of stool culture during the period of August 1981 to July 1982 at Yonsei Medical Center was analyzed and the following results were obtained. 1. C. fetus subsp. jejuni was isolated from 0.8% of stool specimens. The isolation rate was lower than that of salmonella(3.3%) and shigella(7.1%). The isolation was most frequent in June and from $\leq$15-year-old patients. 2. All of the isolates from the patients were susceptible to chloramphenicol and erythromycin. It was noteworthy that 4 isolates were resistant to all of the aminoglycosides, i.e., amikacin, gen tamicin, kanamycin and tobramycin. 3. We also isolated C. fetus subsp. jejuni from chicken. When the susceptibility of the isolates was compared to that of the isolates from human the former were less susceptible to erythromycin(34.1%) and tetracycline(38.6%).

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Outbreaks of Strangles due to Capnophilic Streptococcus equi subsp equi in South Korea (호이산화탄소성 Streptococcus equi subspecies equi에 의한 선역의 발생)

  • Moon, Ja-Ho;Kim, Jeong-Hee;Takai, Shinji;Son, Won-Geun
    • Journal of Veterinary Clinics
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    • v.32 no.1
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    • pp.41-44
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    • 2015
  • We reported an outbreak of clinical strangles in thoroughbred horses due to capnophilic Streptococcus equi subsp equi in South Korea. On three different farms, we isolated 17 S equi subsp equi isolates from 29 horses with or without abscesses in their lymph nodes. Of the 17 isolates, two isolates from clinical cases grew well in aerobic conditions, whereas 7/7 isolates from clinical cases and 8/22 isolates from the nasal discharges of horses did not. The latter 15 isolates were capnophilic, oxygen-sensitive, and $CO_2$-requiring S equi subsp equi, which could not grow in aerobic conditions, but which grew well in a $CO_2$ incubator with 5% $CO_2$, in anaerobic conditions using a GasPak, and with reduced oxygen tension in a candle jar. This study is the first report of a strangles outbreak caused by capnophilic S equi subsp equi in South Korea.

Isolation of Anaerobic Bacteria from Clinical Specimens in Veterinary Medical Teaching Hospital (대학 동물병원 임상 검체로부터 분리된 혐기성 세균과 항생제 감수성 양상)

  • Park, Se-won;Chung, Jin-young;Hwang, Cheol-yong;Youn, Hwa-young;Han, Hong-ryul
    • Journal of Veterinary Clinics
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    • v.21 no.1
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    • pp.15-19
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    • 2004
  • The emergence of bacterial resistance to antibiotics during therapy is a matter of great problem in clinical medicine. This may be because many veterinarians have used inappropriate antibiotics without bacteriological culture. Therefore, the purpose of this study is to determine isolation of anaerobic bacteria as pathogens from veterinary clinical specimens as well as susceptibility pattern for choosing antibiotics. Various anaerobic bacteria were isolated from clinical specimens of dogs, cats, rabbits at Veterinary Medical Teaching Hospital of Seoul National University from May 2001 to October 2002. The total number of isolated anaerobic bacteria was 13 isolates; Bacteroides spp. (3 isolates), Fusobacterium spp. (2 isolates), Peptostreptococcus spp. (2 isolates), Porphyromonas gingivalis (2 isolates), Prevotella spp. (3 isolates), and Propionibacterium acnes (1 isolate). For evaluating the antibiotic susceptibility patterns of the isolates disk diffusion method was used. All isolates were susceptable to all tested antibiotics except only one Fusobacterium varium was resistant to norfloxacin.