• 제목/요약/키워드: Pulsed field gel electrophoresis

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서울시내 수계시설에서 분리된 Legionella spp.의 병원성에 대한 분자역학적 연관성 (Molecular Epidemiological Relationship of the Pathogenicity of Legionella spp. Isolated from Water Systems in Seoul)

  • 김진아;정지헌;김수진;진영희;오영희;한기영
    • 미생물학회지
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    • 제45권2호
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    • pp.126-132
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    • 2009
  • 레지오넬라균은 치명적인 폐렴을 일으킬 수 있는 균으로서, 기후 온난화와 함께 노출의 수위가 높아지고 있는 병원성균이다. 본 연구는 2008년 6월부터 8월까지 서울시내 25개구 소재 수계시설에서 분리한 레지오넬라균 73개 주에 대해 혈청형, Dot/Icm라 일컬어지는 병원성 유전자 분석 그리고 Pulsed field gel electrophoresis (PFGE)를 실시하여 유전자형과 그들 사이의 상관관계를 파악하였다. 73개주 중 69주는 Legionella pneumophila로 혈청형의 분포는 sg1 43주, sg6 9주, sg5 8주, sg3 8주, sg2 1주로 파악되었으며, Legionella spp. 4개주는 Legionella nautarum였다. 분리된 Legionella pneumophila 대부분이 여러 개의 병원성 유전자를 보유하고 있었으며, 반면에 Legionella nautarum은 병원성 유전자가 많이 결핍되어있었다. PFGE pattern을 분석해 볼 때, Legionella pneumophila가 동일한 혈청형안에서 다양하게 분화되어감을 볼 수 있었으며, 병원성 유전자의 분포와 깊은 상관관계를 보였다.

Escherichia coli Genotype을 이용한 생물학적 Critical Control Point의 적합성 평가 (Evaluation of Biological Critical Control Points Using Escherichia coli Genotyping)

  • 김학재;한태욱;정지헌;박경진;홍종해
    • 한국축산식품학회지
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    • 제29권6호
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    • pp.695-701
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    • 2009
  • 본 연구는 축산물가공장 작업환경에 설정된 CCP의 유효성을 평가하기 위하여, 원료도체에서부터 포장에 이르는 전 공정에서 Escherichia coli를 검출하고 PFGE genotyping으로 genetic strain의 흐름을 분석하여 CCP의 적합성을 평가하였다. 시료채취를 위한 작업장은 도축장에서 냉장차량으로 원료도체를 공급받는 독립형 소규모 HACCP 가공장(100두 처리/일)과 축산물종합처리장내 HACCP 가공장(500두 처리/일)을 각각 1곳씩 선정하였다. 시료는 원료도체, 시설 및 장비(발골기, 박피기, 작업대, 벨트, 칼, 면장갑), 환경(벽, 바닥)과 작업공정상의 발골육, 부분육, 최종육을 무균적으로 채취하였다. 작업공정에서 E. coli의 높은 분리율(59.6%)과 작업환경 전반에서의 고른 검출로 E. coli가 축산물가공장 위생지표균으로의 활용 가능성을 확인할 수 있었다. 분리된 E. coli의 genotype 분포는 작업장내 미생물 오염의 흐름을 보여주었다. 소규모가공장의 경우 오염된 원료육 반입이 작업공정을 오염시키고 공정중의 교차오염 발생으로 최종육에 영향을 주었고, 축산물종합처리장내 작업장은 원료육 오염보다는 작업환경에 상존하는 오염미생물의 교차오염이 최종육에 영향을 주는 양상이었다. 따라서 포장육가공장과 같이 작업공정에 멸균처리 과정이 없는 경우는 선행요건에 해당되는 원료반입과 작업환경의 세척 및 소독이 CCP로 관리되거나 이에 준하는 특별관리가 필요하였다. 결과적으로, 작업공정에서의 E. coli 검출과 genotyping은 작업공정상의 미생물 오염의 흐름을 파악하는데 유용하였으며, 이러한 방법은 HACCP 작업장의 미생물 오염원 분석과 설정된 생물학적 critical control point의 적합성 평가에 효과적이었다.

우리나라에서 분리된 참다래 꽃썩음병 병원세균(Pseudomonas syringae pv. syringae)의 플라스미드와 Cu 저항성 유전자 (Plasmid Profiles of Pseudomonas syringae pv. syringae Isolated from Kiwifruit Plants in Korea and the Copper Resistance Determinant)

  • 박소연;한효심;이영선;고영진;신종섭;정재성
    • 미생물학회지
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    • 제43권4호
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    • pp.337-340
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    • 2007
  • Pseudomonas syringae pv. syringae는 우리나라에서 참다래 꽃썩음병의 원인세균으로 알려져 있다. 본 연구에서는 우리나라의 서로 다른 참다래 과수원에서 분리되어 동정된 11개 균주의 꽃썩음병균이 가지고 있는 플라스미드 양상을 pulsed-field 젤 전기영동으로 조사하였다. 그 결과 전체 균주들은 가지고 있는 플라스미드의 개수와 크기에 따라 6개 그룹으로 나누어 졌다. 플라스미드의 수는 0에서 4개, 크기는 22 kb에서 160 kb로 다양하였다. 이들 중 두 개의 플라스미드를 가지고 있는 그를 III에 속하는 4균주가 Cu에 대한 저항성을 보였다. Southern blot hybridization 결과 Cu 저항성 유전자는 48 kb 크기의 플라스미드에 들어 있었다.

중환자실 입원 환자의 비강 도말에서 메티실린 내성 황색포도알균의 분자역학, 항생제 내성 연구 (Molecular Epidemiology and Antimicrobial Resistance of Methicillin-resistant Staphylococcus aureus Isolated from Nasal Swab at Intensive Care Unit)

  • 곽엄섭;권미혜;정지현;강미일;천지영;이고은;김영근;최유진;나문준;권희욱;손지웅
    • Tuberculosis and Respiratory Diseases
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    • 제65권2호
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    • pp.91-98
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    • 2008
  • 연구배경: 메티실린 내성 황색포도알균은 의료기관 관련 감염 뿐 아니라 지역사회 감염에서도 나타나 이에 저자들은 한 대학병원 중환자실에 입원한 환자의 비강 도말 배양을 통해 MRSA의 의료기관 관련 감염 및 지역사회 감염의 빈도와 분자 역학 및 항생제 내성을 연구하였다. 방 법: 2006년 6월에서 9월까지 건양대학교 병원 중환자실 환자 353명을 대상으로 입실 첫날 비강 도말 배양을 시행하여 MRSA 획득 위험인자에 따라 HA-MRSA와 CA-MRSA로 나누어 Pulsed-Field Gel Electrophorosis (PFGE)로 분류하여 각각의 항생제 내성 검사를 시행하였다. 결 과: 353명 중 비강 도말 배양에서 동정된 MRSA는 42명(11.9%)이며, 동정된 MRSA 중 HA-MRSA는 33명(78.6%), CA-MRSA는 9명(21.4%)이다. PFGE에서 type A에서 type K까지 11형으로 구분하였고 HA-MRSA는 type A (n=9), B (n=7)가, CA-MRSA는 type A (n=2), B (n=2)가 주로 나타났다. 항생제 내성률은 erythromycin, ciprofloxacin에서 HA-MRSA가 CA-MRSA보다 높게 나타났다. 결 론: MRSA의 집락률은 11.9%이며 HA-MRSA의 균주가 CA-MRSA보다 많고, CA-MRSA는 9예로 적은 예지만 PFGE type에서 대부분의 type이 HA-MRSA에서 동정된 type과 같은 경향을 보여 지역사회전파를 시사한다.

Molecular Typing and Antimicrobial Susceptibility of Staphylococcus aureus Strains Isolated from Raw Milk, Cheese, Minced Meat, and Chicken Meat Samples

  • Can, Hayriye Yesim;Elmali, Mehmet;Karagoz, Alper
    • 한국축산식품학회지
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    • 제37권2호
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    • pp.175-180
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    • 2017
  • The objectives of this study were: i) to detect the presence of Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) in raw milk, cheese, beef minced meat, and chicken meat samples; ii) to evaluate the antimicrobial susceptibility of the isolates; and iii) to determine clonal relation among the isolates by using pulsed-field gel electrophoresis (PFGE) method. Therefore, a total of 160 food samples were randomly collected between August 2014 and May 2015 in Hatay province, located in the southern Turkey. Twenty (12.5%) of the samples were found to be contaminated with S. aureus. A total of 40 isolates from the 20 positive samples were confirmed to be S. aureus by multiplex PCR based on 16S rRNA and nuc gene. The mec A gene was not detected in any of the S. aureus strains. In the present study, 39 out of 40 (97.5%) isolates were found to be resistant to one or more antibiotics. All of isolates were susceptible to gentamicin, oxacillin, and vancomycin. The highest resistance rate was detected in penicillin (95%) and ampicillin (92.5%), followed by tetracycline (30%), erythromycin (20%), ciprofloxacin (12.5%). Nine major patterns were determined by PFGE. In 6 of these patterns, thirty-six strains (90%) had identical PFGE profiles.

Detection of CTX-M and Clonal Analyses using MLST in Cefotaxime Resistant $Escherichia$ $coli$ Isolated from the Han-River, Korea

  • Son, Gun-Woo;Kim, Mal-Nam
    • 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.

Genomic Relationship of Salmonella enterica Serovar Typhimurium DT104 Isolates from Korea and the United States

  • Kim, Shukho;Chun, Sung-Guen;Lim, Ok-Young;Park, Mi-Sun;Kang, Yeon-Ho;Park, Yong-Ho;Lee, Bok-Kwon
    • Journal of Microbiology
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    • 제42권1호
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    • pp.14-19
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    • 2004
  • Salmonella enterica serovar Typhimurium DT104 (Salmonella Typhimurium DT104 or DT104) has been emerging as a common pathogen for human in Korea since 1997. In order to compare the genomic relationship and to search for the dominant strains in Korea, we conducted pulsed-field gel electrophoresis (PFGE) and IS200 fingerprinting of 25 epidemiological unrelated isolates from human and animals from Korea and cattle from America. Two Salmonella Typhimurium DT104 isolates from human in Korea and all 8 isolates from American cattle had indistinguishable patterns from the PFGE and IS200 fingerprinting but multidrug-resistant Salmonella Typhimurium isolates, including DT104, from Korean animals had diverse genetic patterns. The data suggest that a dominant DT104 strain might have circulated between Korean and American cattle and that it had a high level of clonality.

국내 임상분리주 Streptococcus pneumoniae의 혈청형에 따른 유전적 상관성 (The Genetic Correlations Among Serotypes and PFGE Patterns of Streptococcus pneumoniae Isolated in Korea)

  • 정경석
    • 한국환경보건학회지
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    • 제30권1호
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    • pp.15-21
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    • 2004
  • In an attempt to analyze the characteristics of domestic pathogenic strains of S. pneumoniae, the basic epidemiological charactristics of pathogenic strains such as their serotypes and frequency of penicillin resistance, and pattern of chromosomal DNA from PFGE(pulsed-field gel electrophoresis) were observed. For this study,56 strains of S. pneumoniae isolated from inpatients and outpatients in the four domestic university hospitals were collected from January to December in 1998. Among those strains, a total of 56 pathogenic strains from blood(39 isolates), cerebrospinal fluid(8 isolates) and other specimen(9 isolates) were selected and isolated. The penicillin resistance frequency of those 56 strains was identified with disk diffusion method with 66.1%. From the invasive strains, predominant serotypes were isolated in the order of 19F(12.5%), 23F(10.7%), 14(10.7%) and 9V(10.7%), totalling 45 percent. This experiment also used PFGE patterns to compare the correlations among genetic subtypes in several serotypes. The DNA fragments digested with Sma I and Apa I were resolved by PFGE. The PFGE patterns digested with Sma I were better than Apa I for analysis. In the DNA fragments digested with Sma 1, PFGE analysis of 56 S. pneumoniae isolates showed 25 different patterns. As a result, serotype was on the whole correlated to PFGE pattern on the ground that each different PFGE pattern by serotype was observed. This study can be utilized not only fur the study of incidence trend of domestic pneumococcal diseases but also as a useful basic data for the development of identification tool and treatment.

Riboprint and Virulence Gene Patterns for Bacillus cereus and Related Species

  • Kim, Young-Rok;Batt, Carl A.
    • Journal of Microbiology and Biotechnology
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    • 제18권6호
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    • pp.1146-1155
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    • 2008
  • A total of 72 Bacillus cereus strains and 5 Bacillus thuringiensis strains were analyzed for their EcoRI ribogroup by ribotyping and for the presence or absence of seven virulence-associated genes. From these 77 strains, 42 distinctive ribogroup were identified using EcoRI, but the two species could not be discriminated by their EcoRI ribogroup. The 77 strains were also examined by PCR for the presence of seven virulence-associated genes, cerAB, pi-plc, entFM, bceT, hblA, hblC, and hblD. All five Bacillus thuringiensis strains were positive for these genes. Although differences in the patterns of virulence genes were observed among the different B. cereus strains, within any given ribogroup the patterns of the seven virulence genes was the same. Pulsed-field gel electrophoresis (PFGE) analysis in combination with available chromosomal maps for a selected group of B. cereus strains revealed significant differences in their chromosome size and the placement of virulence genes. Evidence for significant rearrangements within the B. cereus chromosome suggests the mechanism through which the pattern of virulence-associated genes varies. The results suggest linkage between ribogroups and virulence gene patterns as well as no apparent containment of the latter within any particular species boundary.

Comparison of Cellular Fatty Acid Composition and Genotypic Analysis of Bifidobacterium longum MK-G7 with Commercial Bifidobacteria Strains

  • Jung, Hoo-Kil;Kim, Eung-Ryool;Ji, Geun-Eog;Park, Jong-Hyun;Cha, Seong-Kwan;Juhn, Suk-Lak
    • Journal of Microbiology and Biotechnology
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    • 제10권2호
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    • pp.143-146
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    • 2000
  • This study was conducted to compare the cellular fatty acid composition and genotypic analysis of Bifidobacterium longum MK-G7 originated from Koreans with other commercial type strains of bifidobacteria. The cellular fatty acid of Bif. longum MK-G7 was shown to be composed of $C_{160FAME},C_{181\;c18DMA},C_{18.1\;CIS9\; FAME},C_{14.0FAME},C_{19\;0cye9,10 DMA},Feature7(C_{17.2 FAME), and Feature 10(C_{181\; Cll/t9/t6 FAME}$. Bif. longum MK-G7 showed 99.9% homology and the highest relatedness with Bif. longum ATCC 15707 type strain. Both Bif. longum MK-G7 and Bif. longum ATCC 15707 showed 153 bp products on RAPD (randomly amplified polymorphic DNA) analysis, however, they showed quite different band patterns on PFGE (pulsed-field gel electrophoresis) analysis. Consequently, our present study showed that Bif. longum MK-G7 was different from any commercial type strains of Bif. longum tested.

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