• 대한임상검사과학회지
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• 제49권4호
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• pp.413-419
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• 2017

폐렴의 한(일개) 환자의 임상검체에서 연속적으로 분리된 Imipenem 내성세균 4균주를 분리하였다. 분리균을 동정하기 위해 Vitek II system의 GN card를 이용하였으며 16S rRNA유전자 염기서열을 기초로 계통학적 분석을 실시하였다. 분리균은 P. aeruginosa (2 strains), P. monteilii (1) 및 P. putida (1) 으로 동정되었다. 분리균들의 항생제에 대한 내성시험은 Vitek II system AST-N225 card를 이용해서 imipenem의 최소억제 농도가 모두 $${\geq_-}8{\mu}g/mL$$을 확인한 후 실험에 사용하였다. ${\beta}-Lactamase$ 유전자의 특이 시발체를 이용하여 증폭한 PCR 산물로 imipenem 내성 유전자형을 결정하였는데 분리된 4균주 모두에서 MBL 유전자를 확인하였으며 2균주의 P. aeruginosa는 MBL유전자중 VIM형과 SHV형 유전자를 그리고 또다른 균주는 VIM형과 OXA group II형 유전자를 동시에 보유하고 있었다. 항생제 감수성 결과에서는 amikacin이 다른 항생제보다 감수성을 보였을 뿐 대체적으로 내성율이 높았다. 균주들간의 역학적 연관성 분석을 위해 ERIC-PCR을 이용한 DNA 지문 분석결과, 분리된 2 균주의 P. aeruginosa는 유사한 균주일 것으로 추정하였으나 DNA band 유형의 상동성은 서로 다른 유형임을 알아 볼 수 있었다. 특이하게 한 환자에게서 imipenem 내성세균이 4균주가 검출 된 것은 이례적이며 동종의 DNA band 유형도 서로 상이하였다.

• 생명과학회지
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• 제20권8호
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• pp.1214-1220
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• 2010

대한민국 순천의 병원 입원 환자의 검체로부터 imipenem 내성 세균을 분리하였다. 54개의 분리균을 16S rRNA 유전자와 gyrB 유전자 염기서열 비교를 기초로 하여 계통분류학적으로 동정하였다. 분리균들은 Pseudomonas aeruginosa (30균주; 55.6%), Acinetobacter baumannii (21; 38.9%), Enterobacter hormaechei (2)와 Pseudomonas putida (2)에 속했다. 22개의 균주가 metallo-$\beta$-lactamase (MBL)를 생산하였으며 종별 구성은 다음과 같다; Acinetobacter baumannii 12균주, Pseudomonas aeruginosa 7균주, P. putida 2균주 그리고 Enterobacter hormaechei 1균주. 분리균들의 항생제 감수성은 디스크 확산법과 Vitek 을 이용하여 조사하였다. IMP 와 VIM 형의 metallo-$\beta$-lactamase를 생산하는 균주들은 OXA 와 SHV 형 $\beta$-lactamase를 생산하는 균주들에 비해 ceftazidime, aztreonam, amikacin과 gentamicin에 대한 내성율이 높았다.

• 대한의생명과학회지
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• 제20권2호
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• pp.77-84
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• 2014

Periodic analysis of local epidemiologic data of prevalent pathogens of blood culture can provide clinicians with relevant information to guide empirical antibiotic therapy. In this study, we analyzed a pattern of change of causative microorganisms and antimicrobial resistance at a tertiary medical center in Chungcheong province from 2003 to 2012, retrospectively. Of 70,258 blood specimens cultured, 6,063 (8.6%) were positive. Among the positive isolates, 95.9% were aerobic or facultative anaerobic bacteria, 0.1% were anaerobes, and 3.9% were fungi. Coagulase-negative Staphylococci (CoNS) (32.9%), Escherichia coli (16.7%), Staphylococcus aureus (9.1%), Klebsiella pneumoniae (6.4%), and ${\alpha}$-hemolytic Streptococcus (5.9%) were commonly isolated bacteria, and Candida albicans (1.4%) was the most commonly isolated fungi. Enterococcus faecium progressively increased but Streptococcus pneumoniae, Acinetobacter baumannii and Proteus species gradually decreased over a period of 10 years. The multidrug-resistant microorganisms such as methicillin-resistant S. aureus (MRSA), vancomycin-resistant enterococci (VRE), cefotaxime-resistant E. coli, imipenem-resistant Pseudomonas aeruginosa (IRPA) and imipenem-resistant A. baumannii (IRAB), were significantly increased. Therefore, there is a need for a more strict control of antibiotics and a more updated guideline for the treatment of bloodstream infection.

• Clinical and Experimental Pediatrics
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• 제56권8호
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• pp.332-337
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• 2013

Purpose: To identify the frequency of bacterial isolates in early-onset neonatal sepsis (EONS) and their antimicrobial resistance pattern. Methods: A retrospective study of EONS was conducted at the Beni Suef University Hospital from September 2008 to September 2012. A case of EONS was defined as an infant who had clinical signs of infection or who was born to a mother with risk factors for infection, and in whom blood culture obtained within 72 hours of life grew a bacterial pathogen. Results: Of 673 neonates screened, there were 138 positive blood cultures (20.5%) (confirmed EONS). Of the recovered isolates, 86.2% were gram-negative pathogens. Klebsiella pneumoniae (42.8%), Enterobacter cloacae (22.5%), and Escherichia coli (13.8%) were the commonest isolated organisms. The most common gram-positive microorganism was Staphylococcus aureus accounting for only 12 isolates (8.7%). All Klebsiella isolates and 93% of Enterobacter isolates were resistant to ampicillin. Gram-negative pathogens had the maximum overall sensitivity to imipenem, cefepime, and ciprofloxacin; whereas, gram-positive isolates were most susceptible to vancomycin, imipenem, and piperacillin. Conclusion: K. pneumoniae was the predominant causative bacteria of EONS followed by E. cloacae and E. coli. There was a high resistance to ampicillin. Imipenem had the maximum overall activity against the causative bacteria. Continuous surveillance is needed to monitor the changing epidemiology of pathogens and antibiotic sensitivity.

• 환경생물
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• 제29권4호
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• pp.286-295
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• 2011

Bacteria resistant to various antibiotics have recently become an issue of the utmost importance. Resistant strains are not uncommon, even in municipal drinking water sources. The health threat posed by resistant, pathogenic bacteria has serious ramifications for both public health and agriculture. In this study, we isolated antibiotic resistant bacteria from water samples from the Han River, Korea, which is contaminated by the wastewater from many industrial complexes, hospitals, agricultural and animal husbandry estates, and from wastewater treatment facilities. We determined the degrees of resistance to various antibiotics exhibited by the isolated strains. The similarities between the isolated $E.$ $coli$ strains were examined, using the pulsed field gel electrophoresis and multilocus sequence typing, in order to trace their origins and to explore the syntechnic adaptations and pathogenicity of the various strains and relate these to their genetic sequence. A total of 25 $E.$ $coli$ strains were isolated from six stations along the Han River. All the 25 strains exhibited resistance to ampicillin. We also investigated resistance to amoxicillin, clavulanic acid, cefazolin, cofoxitin, cefotaxime, cefpodoxime, ceftriaxone, cefepime, nalidixic acid, aztreonam, ciprofloxacin, streptomycin, gentamicin, chloramphenicol and imipenem. Based on the ESBL detection, 14 strains belonged to the ESBL producing strains. The number of the clonal complex producing strains was 5 among the 14 isolated strains. The 5 strains were included in the 168, 23, 38, 469, 156 clonal complex, respectively. The rest 9 strains were not included in the clonal complex, but showed independent STs.

• 생명과학회지
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• 제22권9호
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• pp.1268-1276
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• 2012

사람의 감염증 치료에 사용되는 carbapenem계 약제에 대한 내성균의 출현 및 확산은 감염증 치료를 제한할 뿐만 아니라 집단 발병의 원인이 될 수 있다. 이에 본 연구에서는 ${\beta}$-lactam 약제에 내성을 갖는 Pseudomonas aeruginosa (P. aeruginosa)를 대상으로 metallo-beta-lactamase (MBL)의 유전형을 규명함으로써 내성세균의 감염증 치료지침 및 확산방지책 마련에 기초 자료를 제공하고자 하였다. 본 연구의 대상이 된 254개의 임상 검체 중에서 42주의 P. aeruginosa 를 분리하여 imipenem 혹은 meropenem에 내성을 나타내는 Hodge 변법과 EDST에서 각각 28주와 23주가 양성반응을 보였다. DNA의 염기서열 분석결과 $bla_{IMP-6}$ 유전자 보유균이 8주, $bla_{VIM-2}$ 유전자 보유균이 17주로 59.5%(25/42)가 MBL을 생성하는 것으로 나타났다. $bla_{IMP-6}$의 유전자 환경은 $bla_{IMP-6}$-qac-aacA4-$bla_{OXA-1}$-aadA1 유전자 배열을 지니고 있었다. 또한 ERIC PCR 결과 IMP-6과 VIM-2를 생성하는 일부 균주에서 역학적 연관성이 있음이 확인되었다. 본 연구에서 분리한 carbapenem계 항균제 내성 P. aeruginosa가 보유한 $bla_{IMP-6}$ 유전자는 대구지역에서 발병이 보고된 유전자의 gene cassette와 일치하는 것으로 확인되었다. 따라서 이들 세균이 지역사회에 정착하고 있고 이들을 보유한 세균에 의한 감염증 치료시 치료약제에 대한 선택압을 증가시킬 것으로 우려된다. 그러므로 항균제 내성 검사를 통하여 적절한 항균제를 선택하고, 항균제 내성균들의 출현과 확산을 막는 연구가 계속되어야 할 것으로 생각된다.

• 대한임상검사과학회지
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• 제51권4호
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• pp.444-452
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• 2019

주요 획득성 metallo-β-lactamase (MBL) 유전자에 의해 매개되는 carbapenem 내성, 특히 Acinetobacter spp. 균종의 임상 분리주에 대한 보고가 증가하고 있다. 본 연구에서 임상에서 비 중복으로 분리된 carbapenem 비감수성 Acinetobacter spp. 191주 중 125 (65.4%)주가 imipenem 혹은 meropenem-Hodge 변법시험에 양성이었고, 49 (25.7%)주가 imipenem-EDTA+SMA double disk synergy (DDS) 시험에 양성이었다. bla_VIM-2 allele와 bla_IMP-2 allele 검출을 위한 중합효소연쇄반응과 염기서열분석을 시행한 결과, A. baumannii와 A. calcoaceticus에서 각각 29주와 1주가 bla_VIM-2를 갖고 있었고, A. baumannii 16주와 A. calcoaceticus 2주가 bla_IMP-1을 갖고 있었다. A. genomospecies 3는 blaVIM-2와 bla_AIM-1을 동시에 갖고 있었다. 이들 MBL 유전자는 모두 class 1 integron에 있었다. bla_VIM-2 혹은 bla_IMP-6를 갖는 class 1 integron의 크기는 A. baumannii 분리주에서는 2.8 kb에서 3.2 kb이었고, A. genomospecies 3 분리주에서는 3.2 kb에서 3.5 kb이었다. bla_VIM-2는 대부분 class 1 integron에 첫번째 혹은 두번째에 위치하였고, aacA4를 흔히 가지고 있었다. 다양한 내성 유전자를 가질 수 있는 MBL 생성 Acinetobacter spp.뿐 아니라 다양한 내성 유전자를 가질 수 있는 integron의 전파로 imipenem이나 meropenem과 같은 carbapenem 내성을 포함하여 다제 내성 그람음성 세균의 증가가 예상된다. 또한, 위중한 Acinetobacter spp. 감염증 치료를 위한 새로운 항균제 개발이 필요하다.

• 한국미생물·생명공학회지
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• 제50권1호
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• pp.147-156
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• 2022

Urinary tract infections (UTIs) are one of the most common infections in different age groups, including children. Bacteria are the main etiological agents of UTIs. The aim of the present study was to isolate, identify, and determine the antibiotic susceptibility of bacteria isolated from children with UTIs from Baghdad, Iraq. Three hundred and two urine samples were collected from children aged 6 months to 12 years. The samples were cultured on blood agar and MacConkey agar. The selected colonies were subjected to biochemical tests and antibiotic susceptibility analysis using the Vitek^® 2 Compact automated microbial identification system. In this sample, 299 bacteria were identified, of which, 267 were gram-negative bacteria, and 32 were gram-positive bacteria. Escherichia coli (56%) was the most commonly isolated gram-negative bacteria, followed by Pseudomonas aeruginosa (14%), Enterobacter spp. (10.48%), Klebsiella pneumoniae (9.36%), Proteus spp. (7.8%), Acinetobacter baumannii (1.5%), and Morganella morganii (0.37%). Enterococcus faecalis (62.5%) was the most commonly detected gram-positive bacteria, followed by Staphylococcus aureus (37.5%). E. coli and P. aeruginosa were the most antibiotic-resistant bacteria. Among the tested antibiotics, meropenem showed 100% sensitivity, followed by imipenem (97.4%), amikacin (91.8%), and tobramycin (83.5%). In contrast, the high frequencies of resistance were observed with cefixime (93.2%), cefotaxime (78.7%), and ceftriaxone/cefotaxime (71.2%). In conclusion, carbapenems and aminoglycosides are highly recommended for the empirical treatment of UTIs, while, Quinolones, penicillins, and cephalosporins are not suggested. Frequent antibiotics susceptibility testing are warranted to determine the resistance pattern of UTI bacteria.

• 한국미생물·생명공학회지
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• 제50권1호
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• pp.89-94
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• 2022

Triclosan (TCS) is an antimicrobial agent used in various human personal care products against both gram-positive and gram-negative bacteria. The purpose of this study was to evaluate the presence of TCS-resistant bacteria in sewage water in Peshawar, Khyber Pakhtunkhwa (KPK), Pakistan, for the first time. TCS-supplemented Luria Bertani (LB) agar was used to isolate TCS-tolerant bacteria. A total of 17 TCS-resistant isolates were randomly selected from a large pool of bacteria that showed growth on TCS-supplemented LB agar. Based on gram staining and physiochemical characteristics, the isolated strains were identified as Salmonella typhi (n = 6), Escherichia coli (n = 4), Citrobacter freundii (n = 4), Proteus mirabilis (n = 1), Enterobacter cloacae (n = 1), and Pseudomonas aeruginosa (n = 1). The Triclosan mean minimum inhibitory concentrations (MICs) for the isolates of Salmonella typhi, Escherichia coli, Citrobacter freundii, Proteus mirabilis, Enterobacter cloacae, and Pseudomonas aeruginosa were 23.66 ㎍ ml^-1, 18.75 ㎍ ml^-1, 42 ㎍ ml^-1, 32 ㎍ ml^-1, 64 ㎍ ml^-1, and 128 ㎍ ml^-1, respectively. The antibiogram revealed that all isolates were resistant to penicillin G (100%) and linezolid (100%), followed by ampicillin (94%), tetracycline (76%), tazobactam (76%), sulbactam/cefoperazone (64%), polymyxin PB (58%), amikacin (29.41%), aztreonam (29.41%), imipenem (5%), and gentamicin (5%). This is the first known study regarding the isolation of TCS-tolerant bacteria from sewage water in Peshawar, KPK, Pakistan. It was concluded that all the TCS-resistant isolates were multidrug resistant (MDR) gram-negative rod-shaped bacteria, mostly belonging to the Enterobacteriaceae family.

• 대한의생명과학회지
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• 제18권3호
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• pp.299-306
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• 2012

Recently, the prevalence of 16S rRNA methylase conferring high-level resistance to aminoglycosides has been increasing in Gram-negative bacilli globally. We determined the prevalence and genotype of these methylase-producing bacteria, and characterized the co-resistance to ${\beta}$-lactam antibiotics and quinolone in Gram-negative clinical isolates collected in 2010 at a hospital in Korea. Among 65 amikacin-resistant isolates screened from 864 Gram-negative bacilli (GNB), 16S rRNA methylase genes were detected from 49 isolates, including Acinetobacter baumannii (43), Klebsiella pneumoniae (2), Proteus mirabilis (2) and Serratia marcescens (1), Empedobacter brevis (1). All of the 16S rRNA methylase genotype was armA and no variant sequences of amplified PCR products for armA were noted. The 16S rRNA methylase producing bacteria showed much higher resistance to aminoglycoside for Enterobacteriaceae and glucose non-fermenting (NF)-GNB and to imipenem for glucose NF-GNB, than the non-producing isolates. All of the 16S rRNA methylase producing Enterobacteriaceae had the extended-spectrum-${\beta}$-lactamase. In addition, two K. pneumoniae concurrently produced both plasmid-mediated AmpC ${\beta}$-lactamase and qnrB gene. All of the amikacin-resistant A. baumannii (43) co-harbored armA 16S rRNA methylase and $bla_{OXA-23}$ carbapenemase. In conclusion, 16S rRNA methylase producing bacteria were very prevalent among GNB in South Korea, and were commonly associated with co-resistance, including carbapenem and quinolone.