• 미생물학회지
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• 제42권2호
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• pp.125-130
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• 2006

본 연구에서는 도축장과 도축물에서 plasmid 매개성 광범위 ${\beta}$-lactam (extended spectrum ${\beta}$-lactamase) 분해효소를 생성하는 Klebsiella pneumoniae와Escherichia coli가 분리됨으로서 한국에서는 이들 균주가 임상범위를 넘어 자연계까지 확산되었음을 확인하였다. 디스크 확산 시험, 등전점 수치, 접합시험 등을 통하여 돼지의 분변으로부터 최종 10균주의 접합자를 얻었고 이들의 형별결정은 아미노산서열분석과 기준주와 대조등을 BCM Search Laucncher 및 NCBI blast를 통해 이루어졌다. 광범위 ${\beta}$-lactam 유형은 Klebsiella pneumoniae 8균주가 TEM-52형이었고 2균주 Escherichia coli가 SHV-12형이었다. CMY-1형은 본 연구에서 분리되지 않았다.

• 미생물학회지
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• 제43권2호
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• pp.116-123
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• 2007

본 연구는 부산지역 하천에서 plasmid 매개성 광범위 ${\beta}-lactam$ 분해효소(extended spectrum ${\beta}-lactamase;\;ESBL$)를 생성하는 세균을 분리하여 생성된 광범위 ${\beta}-lactam$ 분해효소를 유형별로 분류하기 위함이다. Escherichia coli 6균주와 Klebsiella pneumoniae 15균주가 피전달균주인 Escherichia coli J53 $Azid^{R}$에 plasmid 매개성 광범위 ${\beta}-lactam$ 분해효소 생성 인자를 전달하였다. PCR 후 얻은 유전인자를 plasmid로 매개된 광범위 ${\beta}-lactam$ 분해효소 유전자의 염기서열 분석결과, E. coli와 K. pneumoniae 유전자를 BCM Search Launcher 및 GenBank nucleotide database를 통하여 검색한 결과 ESBL 유형은 TEM-52형과 SHV-12형이었다. TEM-52는 E. coli와 K. pneumoniae 양쪽에서 분리되었다. 그러나 SHV-12는 K. pneumoniae에서만 분리되었다. 이상의 결과로부터 plasmid 매개성 광범위 ${\beta}-lactam$ 분해효소를 생성하는 세균이 한국에서는 이미 임상범위를 넘어 자연계까지 확산되었음을 알 수 있었다.

• 미생물학회지
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• 제37권4호
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• pp.277-283
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• 2001

본 논문은 임상검체 이외의 자연계내에서 extended-spectrum $\beta$-lactamase (ESBL) 생성균주의 분리 여부를 확인하고, 분리되었을 경우 이들의 유형을 조사하기 위함이었다. 하수종말처리장, 가물치양식장, 부경대학교 담수어양식장, 공중목욕탕으로부터 분리된 균주를 double disk synergy 검사, 교환접합시험, 등전점 조사 등을 실시하여 하수종말처리장 방류수로붙 extended-spectrum $\beta$-lactamase (ESBL) 생성균주가 26 균주 분리되었다. 균종은 Entero-bacter cloacae와 E. sakazakii, Klebsiella pneumoniae subsp. pneumoniae 및 K. pneumoniae subsp. ozaenae 였다. 이들은 모두 PCR결과 TEM+SHV 복합형이었고 등전점 결과 Klebsiella속의 균들은 pI 5.9, 5.9+5.4 E. cloacae는 $pI{\ge}8.5$, 8.0+5.4, E. sakazakii는 8.0+5.4, 8.0+5.9+5.4였다. 교차접합시험(transconjugation) 결과 피전달 균주인 E. coli RG176 nal$_{r}$에게 K. pneumoniae subsp. pneumoniae (1 균주) K. pneumoniae subsp. ozaenea (5 균주)및 E. cloacae (1 균주)의 $\beta$-lactamase 생성유전자가 전달되었다. 이 결과는 국내에서 임상검체 이외의 자연계에서 분리된 ESBL생성 장내세균의 첫 번째 보고이다.

• 대한의생명과학회지
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• 제12권3호
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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.

• 대한임상검사과학회지
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• 제37권3호
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• pp.149-154
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• 2005

대학병원의 임상검체에서 extended-spectrum-${\beta}$-lactamase (ESBL)생성 Klebsiella pneumoniae 51균주를 분리하였다. ESBL생성 K. pneumoniae 51균주는 광범위한 항생제에 내성을 보였고 대부분의 균주는 amikacin, gentamycin과 ciprofloxacin항생제에 내성을 나타내었다. 대학병원에서 분리한 51균주를 randomly amplified polymorphic DNA (RAPD)로 분석한 결과 충남대학병원 21균주와 충북대학병원에서 분리한 10균주는 Ia 와 Ib에 속하였고 경상대학 병원에서 분리한 20균주는 IIa, IIb에 속하였다. ESBL 생성 K. pneumoniae 51균주는 RAPD 분석으로 4가지의 유전형으로 구분 할 수 있었고 유전적으로 다양하였다. 이상의 결과로 RAPD 분석은 유전형분석에 빠르고 단순하고 경제적인 방법임을 알 수 있었다.

• 대한임상검사과학회지
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• 제38권3호
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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.

• 대한임상검사과학회지
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• 제39권3호
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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.

• 한국임상수의학회지
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• 제27권2호
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• pp.125-129
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• 2010

2년령 암컷 포메라니안이 교통사고로 인한 골반골절로 내원하여 골절 교정수술을 받았다. 그러나 수술 후 술부에서 감염으로 인한 화농성 삼출물이 발생하였고, 분리한 세균의 항생제 감수성 검사 결과 penicillins, cephalosporins, aminoglycosides, quinolones, 그리고 trimethoprim/sulfamethoxazole에 내성이 관찰되었다. 분리한 세균의 동정 및 extended-spectrum $\beta$-lactamase (ESBL) 확진시험을 통해 ESBL 생성 Klebsiella pneumoniae임을 확인하였다. 치료를 위한 Carbapenem계 항생제의 감수성 시험 결과에 따라 meropenem을 선택하여 치료에 이용하였다. 분리된 세균에서 ESBL 유전자의 분자생물학적 검사 결과 TEM-1 ESBL 유전자가 있음을 확인하였으며, 보호자의 손바닥에서 분리된 세균에서도 TEM-1, SHV-1 ESBL 유전자가 검출되었다.

• 대한임상검사과학회지
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• 제38권1호
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• pp.26-33
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• 2006

Recently, the rapid increase in extended-spectrum ${\beta}$-lactamase (ESBL) producing clinical isolates has become a serious problem. In this study, the epidemiologic features and molecular characteristics of ESBL among clinical isolates of Escherichia coli and Klebsiella pneumoniae, antibiotic susceptibility testing, genotype of the ESBL and patterns of chromosomal DNA from PFGE (pulsed field gel electrophoresis) were observed. A total of 53 ESBL-producing clinical isolates (30 of E. coli and 23 of Klebsiella pneumoniae) were collected from two university hospitals in the period of June to July in 2002 and 2003 respectively. The antibiotic resistance frequency of those 53 strains was tested by the disk agar diffusion method with the result that all the strains were resistant to cephalothin. To other antibiotics, the resistance rates of E. coli (30 isolates) were in order of ceftazidime (90.0%), cefotaxime and aztreonam (respectively 83.3%). Also, the resistance rates of K. pneumoniae (23 isolates) were in order of aztreonam (78.3%), ceftazidime (73.9%) and cefotaxime (65.3%). Also the sensitivity of ceftazidime-clavulanic acid were 100% in E. coli and 95.7% in K. pneumoniae. And the sensitivity of cefotaxime-clavulanic acid was 96.7% in E. coli and 91.3% in K. pneumoniae. The types of the ESBL genes were determined by using polymerase chain reaction (PCR). Among the 30 isolates of ESBL-producing E. coli, 6 (20.0%) have SHV only, 5 (16.7%) have TEM only and, 18 (60.0%) have both of TEM and SHV. Among the 23 isolates of ESBL-producing K. pneumoniae, 7 (30.4%) have SHV only, 2 (8.7%) have TEM only, and 14 (60.9%) have both of TEM and SHV. These results show that 52 strains, with only one exception, were confirmed as either TEM or SHV. The patterns of Xba I-digested chromosomal DNA of ESBL-producing E. coli and K. pneumoniae isolates were analyzed by PFGE. PFGE patterns of E. coli and K. pneumoniae were multiclonal, but many strains were grouped into a few types. Therefore, it seems that there were clonal outbreaks or possible horizontal spread. In conclusion, the TEM and SHV ${\beta}$-lactamase are most widely spread in E. coli and K. pneumoniae in Korea. As these types are usually carried by plasmids, the spread of these ${\beta}$-lactamase genes could compromise the future usefulness of third generation cephalosporins for the treatment of infections caused by E. coli and K. pneumoniae.

• Journal of Microbiology and Biotechnology
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• 제19권9호
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• pp.1065-1069
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• 2009

Of 143 clinical isolates of Klebsiella pneumoniae collected from Korean non-tertiary hospitals, 24 (16.8%) showed an extended-spectrum ${\beta}$-lactamase-positive phenotype. PCR and sequence analysis revealed the presence of TEM-116 (n=13), CTX-M-3 (n=5), CTX-M-14 (n=2), CTX-M-15 (n=3), and SHV-12 (n=16). Each of the 24 isolates encoded more than one ${\beta}$-lactamase, and seven isolates (29%) harbored two different SHV-type ${\beta}$-lactamase genes ($bla_{SHV-11}$ and $bla_{SHV-12}$) bounded by insertion sequence IS26 in a single transferable plasmid.