• Journal of Veterinary Science
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• 제23권3호
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• pp.37.1-37.8
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• 2022

Background: Avian pathogenic Escherichia coli (APEC) causes colibacillosis, resulting in significant economic losses in the poultry industry. Objectives: In this study, the molecular characteristics of two extended-spectrum beta-lactamase (ESBL)-producing APEC isolates were compared with previously reported ESBL-producing E. coli isolates. Methods: The molecular characteristics of E. coli isolates and the genetic environments of the ESBL genes were investigated using whole genome sequencing. Results: The two ESBL-producing APEC were classified into the phylogenetic groups C and B1 and ST410 and ST162, respectively. Moreover, the ESBL genes of the two isolates were harbored in different Inc plasmids. The EC1809182 strain, harboring the bla_CTX-M-55 gene on the plasmid, exhibited extensive homology to IncFIB (98.4%) and IncFIC(FII) (95.8%). The EC1809191 strain, harboring the bla_CTX-M-1 gene, was homologous to IncI1-I (Gamma) (99.3%). All chromosomes carried the multidrug transporter, mdf(A) gene. Mobile genetic elements, adjacent to CTX-M genes, facilitated the dissemination of genes in the two isolates, analogous to other ESBL-producing E. coli isolates. Conclusions: This study clarifies the transmission dynamics of CTX-M genes and supports strengthened surveillance to prevent the transmission of the antimicrobial-resistant genes to humans via the food chain.

• Journal of Microbiology and Biotechnology
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• 제26권9호
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• pp.1643-1649
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• 2016

The aims of this study were to characterize the molecular epidemiological profiles of CTX-M-producing uropathogenic Escherichia coli isolates from a tertiary hospital in Daejeon, Korea, and to investigate the genetic diversity and compare the prevalence of sequence types (STs) in different areas. Extended spectrum β-lactamase-producing E. coli strains isolated from urine were analyzed for CTX-M, integrons, and insertion sequence common regions (ISCRs) by PCR and sequencing. Multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), phylogenetic analysis, and rep-PCR were also used for molecular typing of the isolates. Of 80 CTX-M producers, 31 and 46 expressed CTX-M-15 and CTX-M-14, respectively. MLST analysis indicated that the most prevalent ST was ST131 (n = 34, 42.5%), followed by ST38 (n = 22, 27.5%), ST405 (n = 8, 10.0%), and ST69 (n = 6, 7.5%). Most CTX-M producers harbored class 1 integrons. ST131 strains belonged to phylogenetic group B2 and showed identical rep-PCR patterns, whereas ST69, ST38, and ST405 strains belonged to phylogenetic group D; the ST38 and ST405 strains displayed the same rep-PCR pattern, respectively. ST131 and ST38 isolates showed 21 and 19 distinct types, respectively, by PFGE. In Daejeon, D-ST38 CTX-M-14 producers were relatively more prevalent than in other countries and Korean cities. Our results indicate that CTX-M-producing E. coli isolates belonged mostly to ST131 or ST38 and were more related to hospital-onset than to community-onset infections and that the bla_CTX-M gene may vary according to the ST.

• Journal of Microbiology and Biotechnology
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• 제27권9호
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• pp.1716-1723
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• 2017

The rapid dissemination of extended-spectrum ${\beta}$-lactamase (ESBL)-producing Escherichia coli has significantly contributed to public health hazard globally. A total of 281 E. coli strains recovered from pigs and chickens between 2009 and 2015 in South Korea were analyzed for ESBL production. ESBL phenotypes were recognized in 14 E. coli isolates; ten and three ESBL-producing isolates carried only $bla_{CTX-M}$ and $bla_{CMY}$ genes, respectively, and one isolate harbored both genes. The predominant CTX-M and CMY types were CTX-M-15 (n = 8) and CMY-2 (n = 3). We also detected ESBL-producing isolates harboring $bla_{CTX-M-65}$, $bla_{CTX-M-14}$, $bla_{CMY-6}$, $bla_{DHA-1}$, and $bla_{TEM-1}$ genes. All ESBL-producing isolates showed resistance to the extent of the fourth generation cephalosporins, along with multidrug resistance. CTX-M-15-producing isolates showed higher MIC values than CTX-M-14- and CTX-M-65-producing isolates. The $bla_{CTX-M}$ and $bla_{CMY}$ genes have the potential to be transferable. The spreading of $bla_{CMY}$ and $bla_{CTX-M}$ genes was arbitrated mainly via Frep and IncI1 plasmids. Our isolates showed clonal diversity in PFGE analysis. This is the first report of E. coli isolates carrying $bla_{CMY-6}$ in chicken from South Korea. The emergence of CMY-6 ESBLs in a population of poultry suggests that extensive screening with long-term surveillance is necessary to prevent the dissemination of ESBL from chicken to human.

• 한국미생물·생명공학회지
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• 제34권4호
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• pp.342-351
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• 2006

병원내 항생제 다제 내성을 일으키는 CTX-M형 ESBL을 생성하는 E. coli와 Klebsielia pneumoniae의 생성현황을 조사하고 이들 균주로 인한 감염증치료와 역학적 조사연구에 도움이 되고자 효소의 유전형을 규명하였다. 2005년 7월-12월에 부산에 소재하고 있는 2개의 종합병원에서 분리된 E. coli와 K. pneumoniae 각각 153주, 52주를 수집하였다. 그 중에서 ESBL을 생성 하는 균주를 검출하기 위해 Double disk synergy test를 시행하여서 E. coli 23주와 K. pneumoniae 13주를 분리하였다. 균주의 동정은 Vitek system GNI card(bioMerieux Vitek Inc., Hazelwood, Mo., U.S.A.)로 확인하였고, 항생제감수성시험은 disk diffusion method 와 agar dilution method를 사용하였다. 분리된 균주들의 내성을 일으키는 ESBL유전형을 규명하기 위하여 Isoelectric focusing(IEF), polymerase chain reaction test, DNA sequencing을 시행하였다. A병원의 13주와 B병원의 10주로 총 23주의 E. coli(15.0%)와 A병원의 7주와 B병원의 6주로 K. pneumoniae 13주(25.0%)가 double disk synergy test 양성으로 ESBL 생성균주로 판정하였다. ESBL 생성 36균주를 대상으로 bla$_{TEM}$, bla$_{SHV}$, bla$_{CTX-M}$ 유전자 검출을 위한 PCR을 시행한 결과 bla$_{TEM}$ 유전자는 13주(36.1%), bla$_{SHV}$ 유전자는 13주(36.1%), bla$_{CTX-M}$ 유전자는 32주(88.9%)가 양성반응을 보여서 bla$_{CTX-M}$ 유전자를 가진 균주가 가장 많이 나타났다. 그리고, bla$_{TME}$, bla$_{SHV}$ 두 가지 유전자를 가지고 있는 균주는 1주(2.8%)만 나타났고 bla$_{TEM}$, bla$_{CTX-M}$두 가지 유전자를 가지고 있는 균주는 9주(25.0%), bla$_{SHV}$, bla$_{CTX-M}$ 두 가지 유전자를 가지고 있는 균주가 10주(27.8%)로 나타나 bla$_{CTX-M}$을 포함하는 복합유전자가 많이 증가함을 알 수 있었다. 또한 CTX-M형 ESBL을 생성하는 E. coli와 K. pneumoniae에 대한 cefutaxime의 MIC는 256 $\mu$g/m1 이상으로 ceftazidime의 16-256 $\mu$g/mL 이상보다 높은 분포를 보였다. 즉, CTX-M형 ESBL 유전자를 지닌 균주에 대한 cefotaxim의 MIC는 ceftazidime의 MIC에 비해서 상대적으로 높은 양상을 보였다. 이러한 결과는 국내의 대학병원 뿐 만 아니라 일반종합병원에서도 CTX-M형 ESBL 생성 E. coli와 K. pneumoniae가 존재하며 확산 중임을 시사한다. 앞으로 CTX-M형 ESBL의 만연과 변종 CTX-M형 ESBL의 출연을 감시하기 위한 정기적인 연구와 조사가 필요한 것으로 생각한다.

• 대한임상검사과학회지
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• 제45권2호
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• pp.43-47
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• 2013

This study was designed to evaluate the prevalence of ESBL genes and monitor antimicrobial resistance pattern in Escherichia coli, isolated from a hospital and a community. We tested 200 E. coli strains isolated in the hospitals and community in Chungcheong area from January to March 2012. Antimicrobial susceptibilities were tested by using the disk diffusion method. A search for ESBL genes was conducted by PCR amplification, and the genotypes were determined by direct nucleotide sequence analysis of the amplified products. An Epidemiologic study was performed by repetitive extragenic palindromic sequence-based PCR (REP-PCR). The percentage of ESBL-producing isolates was 17% for hospital associated E. coli and 11% for community associated E. coli. The ESBL gene sequencing results showed that the most common ESBL in E. coli was CTX-M-14 (19/28), followed by CTX-M-15 (9/28). The REP-PCR study also showed the genetic diversity, but there was no difference between the hospital and community associated E. coli. In this study, the most common types of class A ESBLs identified were CTX-M in the hospital and the community in Chungcheong area. ESBL-producing E. coli isolates showed diverse clonality.

• Journal of Microbiology and Biotechnology
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• 제24권3호
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• pp.421-426
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• 2014

To characterize the extended-spectrum beta-lactamases (ESBLs) in diarrheagenic Escherichia coli from Korea in 2008-2011, we screened seven enterotoxigenic E. coli (ETEC) and one enteroaggregative E. coli (EAEC) that produce ESBLs from a nationwide survey. All eight isolates produced CTX-M-type ESBLs, including CTX-M-12 (n = 4), CTX-M-14 (n = 2), and CTX-M-15 (n = 2). PCR-based replicon typing indicated that the $bla_{CTX-M-12}$ genes of four ETEC isolates were carried on a conjugative IncF plasmid, whereas the $bla_{CTX-M-14}$ of one EAEC was located on an IncK plasmid. This is the first report of the occurrence of $bla_{CTX-M}$ genes in clinical isolates of EAEC in Korea. The ESBL-producing isolates were shown to be different based on pulsed-field gel electrophoresis and multilocus sequence typing, whereas the four isolates with CTX-M-12 were clonally related. These observations raise an alarm for the spread of plasmid-mediated resistance to ESBL among diarrheagenic E. coli.

• 생명과학회지
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• 제23권5호
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• pp.703-709
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• 2013

본 연구에서는 ESBL을 생성하는 Escherichia coli의 Extended-spectrum ${\beta}$-lactamase (ESBL) 유전자와 퀴놀론 내성결정부위(qnr)의 유전자형과 항생제 내성 양상을 규명하고자 하였다. 임상검체에서 분리 된 E. coli 274개 균주를 대상으로 double-disk synergy test 검사를 실시하여 42개의 ESBL 생성 균주를 분리하였다. 검체별로는 소변에서 28균주가 분리되었으며, 객담에서 6균주, 농에서 3균주, 상처에서 2균주, 혈액에서 2균주 그리고 조직에서 1균주가 분리되었다. 이 균주들을 대상으로 ESBL 유전자와 퀴놀론 내성 유전자를 PCR을 이용하여 검색하였다. 35개의 균주가 1개 혹은 2개의 ESBL 유전자를 보유하고 있었다. ESBL 유전자의 분포는 CTX-M-1이 가장 많았으며, CTX-M-9과 TEM 유전자 순이었다. SHV, CTX-M-2와 CTX-M-8는 검출되지 않았다. qnr 유전자는 10개 균주에서 검출되었으며 유전형별로는 qnrB4, qnrB1, qnrS 1 순이었다. 2가지 이상의 ESBL 유전자를 동시에 보유한 균주와 ESBL과 qnr 유전자를 동시에 보유한 균주가 검출되었다. ESBL 유전자 보유균주는 cefotaxmie (80.0%), levofloxacin (82.9%)과 ampicillin (100%)에 고도내성을 보였다. qnr 유전자 보유 균주의 cefotaxmie, levofloxacin과 ampicillin에 대한 내성율은 각각 70%, 70%, 100%였다. ESBL 유전자들간의 그리고 qnr 유전자와의 동시 보유가 항생제 내성에 미치는 상승효과는 없었다. qnr 유전자 보유와 퀴놀론에 대한 내성 사이의 상관관계도 없었다.

• 대한임상검사과학회지
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• 제48권3호
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• pp.210-216
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• 2016

최근 들어 CTX-M형 extended-spectrum ${\beta}$-lactamase(ESBL) 생성 대장균이 국내는 물론 전세계적으로 빠르게 증가하고 있다. 본 연구에서는 충청지역에 위치한 일개의 대학병원에서 분리된 대장균을 대상으로 ESBL 유전자를 중합효소연쇄반응 및 염기서열 분석방법을 통해 확인하였으며, 같은 방법으로 ESBL 생성 대장균으로부터 plasmid mediated quinolone resistance (PMQR) 유전자의 빈도를 조사하였다. 16.0%에 해당하는 25균주가 CTX-M-14를 생성하였으며 이중 9균주는 CTX-M-15도 동시에 생성하는 것으로 나타났다. 항균제 감수성 시험결과 CTX-M형 ESBL을 생성하는 대장균은 모두 cefotaxime에 내성을 보였다. 한편 CTX-M형 ESBL을 생성하는 대장균의 48% (12균주)가 PMQR 유전자를 포함하고 있음이 확인되었는데 8균주가 qnrS1유전자를 그리고 8균주가 aac(6')-Ib-cr 유전자를 포함하고 있었다. 그 중 4균주는 두 개의 유전자를 모두 가지고 있는 것으로 나타났다. 본 연구에서는 플라스미드를 통해 확산될 수 있는 ESBL 및 PMQR 유전자가 대장균 사이에 확산되어 있음을 확인하였다. 항균제 내성유전자들의 확산을 막기 위해서는 지속적인 내성유전자의 모니터링과 감시가 필요할 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제24권6호
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• pp.765-770
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• 2014

This study aimed to characterize CTX-M producers of urinary E. coli and K. pneumoniae isolates and to determine the prevalence of plasmid-mediated antimicrobial resistance genes among them. Minimum inhibitory concentrations (MICs) were determined, and PCR and sequencing were performed. Among the 42 (82.3%) E. coli and 24 (77.4%) K. pneumoniae isolates containing $bla_{CTX-M}$, $bla_{CTX-M-14}$ and $bla_{CTX-M-15}$ were detected in 23 and 19 E. coli isolates, respectively, and in 7 and 17 K. pneumoniae isolates, respectively. CTX-M producers of urinary E. coli and K. pneumoniae were resistant to multiple antibiotics and contained other antimicrobial resistance genes. CTX-M-15 producers contained more antimicrobial resistance genes than did CTX-M-14 producers.

• 한국동물위생학회지
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• 제34권1호
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• pp.37-43
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• 2011

This study was conducted to investigate the prevalence and genotypes of extended-spectrum ${\beta}$-lactamase (ESBL) in 377 Escherichia coli isolated from healthy and sick animals. Two isolates (0.5%), each of which were isolated from diseased swine and chicken, respectively, were confirmed as ESBL producing isolates by double disk synergy test, and showed a multidrug resistant phenotype. Minimum inhibitory concentration of cefotaxime for the two ESBL producing isolates were 3~4 times higher than those of ceftazidime, respectively. By PCR and sequencing, one isolate from swine have both $bla_{CTX-15}$ and $bla_{TEM-1}$, and one isolate from chicken have $bla_{CTX-15}$ and $bla_{TEM-116}$. Also, these genes were transferred to E. coli J53 by conjugation. These two isolates showed unrelated pulsed-field gel electrophoresis. To our knowledge, this is the first time that $bla_{TEM-116}$ gene was identified in E. coli isolated from animals in Korea. These results suggest more prudent use of third- generation cephalosporins, and surveillance and monitoring for ESBL producing E. coli in both animals and their environments should be necessary.