• Title/Summary/Keyword: Pulsed-field-gel-electrophoresis

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

  • Kim, Jin-Ah;Jung, Ji-Hun;Kim, Soo-Jin;Jin, Young-Hee;Oh, Young-Hee;Han, Gi-Young
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.126-132
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    • 2009
  • Legionella spp. is the causative agent of Legionellosis, which induces a potentially fatal form of pneumonia. With a concentrated performance during the summer of 2008, we secured 73 isolates from the water systems of 25 wards in Seoul. We analysed serotypes, pathogenic genes (Dot/Icm), and patterns of pulsed-field gel electrophoresis (PFGE) in an attempt to confirm relationships among them. Different from the previous year's pattern (2007), among the isolates, 69 were Legionella pneumophila and 4 were Legionella spp. The serotype distribution of Legionella pneumophila was sg1 43, sg6 9, sg5 8, sg3 8, and sg2 1. The serotype for the 4 Legionella spp. was Legionella nautarum. Most of the Legionella pneumophila had several pathogenic genes. On the other hand, the 4 Legionella spp. were defective in pathogenicity in genomic terms. The PFGE patterns of the serotypes showed a tendency for diversity of Legionella pneumophila and a close correlation with genetic pathogenicity.

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

  • Kim, Hak-Jae;Hahn, Tae-Wook;Juong, Ji-Hun;Bahk, Gyung-Jin;Hong, Chong-Hae
    • Food Science of Animal Resources
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    • v.29 no.6
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    • pp.695-701
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    • 2009
  • This study was performed to evaluate the effectiveness of biological critical control points using the genetic profile of Escherichia coli isolates from pork cutting plants. Samples were collected from carcasses, equipment (knife, table, glove, transport belt, boning and skinning machine), the environment (wall and floor), and meat cuts during the cutting process from two plants. Pulsed-field gel electrophoresis (PFGE) was used to characterize the E. coli isolates. An identical genotype was detected from the carcasses, equipment, environment, and final meat cuts, and showed that the incoming carcasses, which were contaminated during transportation from slaughterhouses, were a major source of E. coli that was spread throughout processing. Also, consistent cross-contamination due to improper cleaning and disinfection procedures was another possibility. As a result, incoming carcasses and cleaning procedures should be considered critical control points in pork cutting plants, since a heating step is not used to inactivate microorganisms. Furthermore, the high rate (59.6%) of E. coli isolation indicates E. coli can be a good indicator in livestock processing plants even though it has genetic diversity.

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

  • Park, So-Yeon;Han, Hyo-Shim;Lee, Young-Sun;Koh, Young-Jin;Shin, Jong-Sup;Jung, Jae-Sung
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.337-340
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    • 2007
  • Pseudomonas syringae Pv. syringae is a causal agent of bacterial blossom blight of kiwifruit in Korea. Eleven strains of the pathogen were isolated from different kiwifruit orchards in Korea and the plasmid profiles were obtained by pulsed-field gel electrophoresis. They could be clustered into six groups according to the number and size of plasmids. The number of plasmids per strain and size of these plasmids ranged from 0 to 4 and from 22 to 160 kb, respectively. Among them, four strains belonging to Group III which harbored two plasmids were resistant to copper sulfate. Southern blot hybridization of the plasmid DNA indicated that the copper resistance determinant was carried on a 48 kb plasmid.

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

  • Kwak, Om Sub;Kwon, Mee Hye;Jeong, Ji Hyun;Kang, Mi-il;Cheun, Ji Young;Lee, Go Eun;Kim, Young Keun;Choi, Eu Gene;Na, Moon Jun;Kwon, Hee Uk;Son, Ji Woong
    • Tuberculosis and Respiratory Diseases
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    • v.65 no.2
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    • pp.91-98
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    • 2008
  • Background: Methicillin-resistant Staphylococcus aureus (MRSA) is the most common organism associated with nosocomial infections. MRSA infections are becoming increasing important because they have emerged no only as healthcare-associated (HA) infections but also as community-associated (CA) ones. This study examined the moleculo-epidemiology of MRSA, which was isolated from nasal swabs in the intensive care unit (ICU) at Konyang University Hospital. MRSA are classified into HA-MRSA and CA-MRSA. Methods: From June to September 2006, 353 patients who were admitted to the ICU in Konyang University Hospital were enrolled in this study. Single nasal swabs were obtained for culture in the ICU on the 1st day. Pulsed-field gel electrophoresis and the antimicrobial resistant patterns were analyzed between HA- and CA-MRSA. An antimicrobial sensitivity test was also performed. Results: Forty two strains of MRSA were isolated from 353 patients (11.9%). Among the 42 isolates, HA-MRSA and CA-MRSA were found in 33 (78.6%), and 9 (21.4%), respectively. Eleven different PFGE types (type A to K) were identified. Types A (n=9) and B (n=7) were the most common for HA-MRSA, and types A (n=2) and B (n=2) were identified in CA-MRSA. The proportion of types A and B in CA-MRSA (44.4%) was similar to that in HA-MRSA (48.5%). The rates of resistance rates to erythromycin and ciprofloxacin were higher in HA-MRSA than in CA-MRSA. Conclusion: The rate of isolation of MRSA in an ICU setting was 11.9%. HA-MRSA was isolated more frequently than CA-MRSA. The rate of resistance of HA-MRSA to erythromycin and ciprofloxacin was higher than that of CA-MRSA. Despite the small number of subjects, the main isolates (type A and B) of CA-MRSA were similar to those of HA-MRSA.

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
    • Food Science of Animal Resources
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    • v.37 no.2
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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
    • Korean Journal of Environmental Biology
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    • v.29 no.4
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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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    • v.42 no.1
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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.

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

  • 정경석
    • Journal of Environmental Health Sciences
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    • v.30 no.1
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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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    • v.18 no.6
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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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    • v.10 no.2
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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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