• Title/Summary/Keyword: Pulsed field gel electrophoresis (PFGE)

Search Result 86, Processing Time 0.022 seconds

Epidemiologic Investigation on Sporadic Occurrence of Shigellosis in a Subcounty of Cheongwon County in Chungbuk Province in 2003 (청원군 일개 면에서 산발적으로 신고한 세균성이질 집단 발병 역학조사)

  • Lee, Yong-Jae;Hwang, Ue-Kyoung;Kim, Jong-Suk;Kim, Jun-Young;Lee, Bok-Kwon;Koo, Ja-Seol;Kang, Jong-Won
    • Journal of Preventive Medicine and Public Health
    • /
    • v.38 no.2
    • /
    • pp.182-188
    • /
    • 2005
  • Objectives: This study was undertaken to investigate the source of infection and mode of transmission of shigellosis, which occurred sporadically among residents and students in a subcounty of Cheongwon county, Chungbuk province, Korea, from June 4 to July 3 2003. Methods: 692 subjects completed a questionnaire and provided a swab for microbiological examinations,and 7 environmental specimens were examined for bacterial organisms. PFGE (pulsed-field gel electrophoresis) and fingerprinting were performed to find the genetic relationship among the temporally associated sporadic isolates. Results: A total of 29 patients had symptoms consistent with the case definition, with 13 confirmed and 16 suspected cases. The frequency of diarrhea was 6 times or more a day (80.8%), with a duration of 1 to 4 days (88.5%) in most cases. The most common symptoms accompanying the diarrhea were fever (80.9%) followed by abdominal pain (76.9%), headache (65.4%), chill (61.5%), vomiting (46.2%) and tenesmus (15.4%). The epidemic curve was characteristic of a person-to-person transmission. The PFGE and fingerprinting demonstrated identical or similar DNA patterns among the 3 Shigella sonnei isolates (A51, A53 and A61 types) found in this outbreak. Conclusion: A genetically identical strain of S. sonnei was estimated to be the cause of this outbreak, and the mode of transmission was most likely person-to-person.

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
    • /
    • v.45 no.2
    • /
    • pp.126-132
    • /
    • 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.

Antimicrobial Susceptibility and Clonal Relatedness between Community- and Hospital-Acquired Methicillin-Resistant Staphylococcus aureus from Blood Cultures

  • Jung Sook-In;Shin Dong-Hyeon;Park Kyeong-Hwa;Shin Jong-Hee
    • Journal of Microbiology
    • /
    • v.44 no.3
    • /
    • pp.336-343
    • /
    • 2006
  • We compared the antimicrobial resistance and clonal relationships among the community-acquired (CA) and hospital-acquired (HA) methicillin-resistant Staphylococcus aureus (MRSA) strains that were isolated from blood cultures in a university hospital over a 4-year period. A total of 131 MRSA isolates, including 28 CA-MRSA and 103 HA-MRSA strains, were identified; antimicrobial susceptibility testing indicated that the CA-MRSA isolates were more susceptible to erythromycin (21 % vs 6% ; P=0.02), clindamycin (46% vs 12%; P<0.01), ciprofloxacin (43% vs 11%; P<0.01), and gentamicin (43% vs 6%; P<0.01) than were the HA-MRSA isolates. Pulsed-field gel electrophoresis (PFGE) typing and antimicrobial resistance profiles separated the 20 CA-MRSA isolates into 14 and 10 different patterns, respectively, and the 53 HA-MRSA isolates were separated into 24 and 7 different patterns, respectively. Twenty-one (40%) of the 53 HA-MRSA isolates belonged to two predominant PFGE types, and most of them showed multi-drug resistant patterns. Four (20%) of the 20 CA-MRSA and 10 (19%) of the 53 HA-MRSA isolates fell into two common PFGE patterns, and each of them showed the same multi-drug resistant pattern. This study suggests that, although the CA-MRSA blood isolates showed diverse PFGE and antimicrobial resistance patterns, some of these isolates may have originated from the HA-MRSA strains.

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
    • /
    • v.37 no.2
    • /
    • pp.175-180
    • /
    • 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.

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
    • /
    • v.42 no.1
    • /
    • pp.14-19
    • /
    • 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.

Comparison of the safety and immunogenicity of commercial S. gallinarum 9R vaccine (국내 시판 Salmonella gallinarum 9R vaccine의 안전성 및 면역원성 비교)

  • Hwang, Jei Kiun;Lee, Young Ju
    • Korean Journal of Veterinary Research
    • /
    • v.49 no.2
    • /
    • pp.127-133
    • /
    • 2009
  • Salmonella enterica subsp. enterica serovar gallinarum (S. gallinarum) is the agent of fowl typhoid, and the 9R vaccine is a commercial live vaccine for the prevention of fowl typhoid. The aim of this study was to assess the safety and immunogenicity of different brands of S. gallinarum 9R vaccine used in commercial laying chickens in Korea. All 9R strains originated from three different brands showed the same pattern in the biochemical and serological properties, and pulsed field gel electrophoresis (PFGE) profile. But, there was a difference in rhamnose fermentaion, agglutination with Salmonella group $D_1$ antiserum and PFGE pattern between 9R vaccine strain and field S. gallinarum isolates. In laboratory and field trials for assesment of safety and immunogenicity of 9R vaccine, all of the three 9R vaccines showed the same safety in commercial laying chickens. In addition, there was a significant difference between the vaccinated and unvaccinated control groups in mortality and the re-isolation rate of the challenge strain from the tissues (p < 0.05), and no difference by the brands of 9R vaccine. The results from this study indicated that all three different brands of S. gallinarum 9R vaccine showed highly protection against mortality and organ invasion in commercial laying chickens exposed to virulent strains of S. gallinarum.

Megabase-sized DNA isolation and electrophoretic karyotype of fusarium oxysporum schlecht

  • Park, Min-Seon;Min, Byung-Re
    • Journal of Microbiology
    • /
    • v.33 no.2
    • /
    • pp.132-135
    • /
    • 1995
  • To investigate the electrophoretic karytype of Fusarium oxysporum, intact chromosomal DNA was separated by pulsed-field gel electrophoresis (PEGE). DNA extraction from nulcei, mycelia and protoplasts were compared with one another and with the quantity and the suitability for PFGE separation in agarose gel. As a result, the most useful extracting method for intact DNA was found to be that from protoplasts. By varying the electrophoretic conditions, 8 chromosomal DNA bounds were resolved. Using the Schizosaccharomyces pombe and Saccharomyces cerevisiae as size standards, the size of Fusarium oxysporum chromosomes was estimated to range from approximately 0.6 Mb TO 6.7 Mb, and total genome size was 26.7 Mb. The suitability of electrophoretic karyotyping as a tool for strain characterization is discussed.

  • PDF

Genetic Diversity of Multi-resistant Salmonella enterica Serotype Typhimurium Isolates from Animals and Humans

  • Woo Yong-Ku;Lee Su-Hwa
    • Journal of Microbiology
    • /
    • v.44 no.1
    • /
    • pp.106-112
    • /
    • 2006
  • In this study, the genetic diversities of multi-resistant Salmonella typhimurium (ST) isolates were analyzed via the application of both pulsed field gel electrophoresis (PFGE) and Polymerase chain reaction (PCR) analysis methods, using 6 kinds of primers (REP, ERIC, SERE, BOX, P-1254 and OPB-17). And their discriminative abilities (DA) were also compared in order to determine the most effective and reliable analysis method. 118 S. typhimurium isolates, cultured from diverse animals and human patients in Korea beginning in 1993, were analyzed and subjected to a comparison of Simpson's index of diversity (SID), using both PFGE and PCR methods. PFGE by XbaI enzyme digestion allowed for discrimination into 9 pulsotypes, with high SID values (0.991) on the genomic DNA level. This shows that PFGE is a very discriminative genotypic tool, and also that multiple clones of S. typhimurium isolates had existed in domestic animals and humans in Korea since 1993. However, we could ultimately not to trace the definitive sources or animal reservoirs of specific S. typhimurium isolates examined in this study. Depending on the SID values, the combined method (7 kinds of method) was found to be the most discriminative method, followed by (in order) SERE-PCR, REP-PCR, ERIC-PCR, PFGE & OPB-17 (RAPD), P-1254 (RAPD), and BOX-PCR at the $80\%$ clone cut-off value. This finding suggests that the REP-PCR method (which utilizes 4 primer types) may be an alternative tool to PFGE for the genotyping of S. typhimurium isolates, with comparable cost, time, and labor requirement. The establishment of a highly reliable and discriminatory method for epidemiologic analysis is considered necessary in order for researchers to trace the sources of specific pathogens and, consequently, to control and prevent the spread of epidemic S. typhimurium isolates to humans.

Comparison of PFGE, IS6110-RFLP, and 24-Locus MIRU-VNTR for Molecular Epidemiologic Typing of Mycobacterium tuberculosis Isolates with Known Epidemic Connections

  • Jeon, Semi;Lim, Nara;Park, Sanghee;Park, Misun;Kim, Seonghan
    • Journal of Microbiology and Biotechnology
    • /
    • v.28 no.2
    • /
    • pp.338-346
    • /
    • 2018
  • Two molecular epidemiologic methods, IS6110 restriction fragment length polymorphism (IS6110-RFLP) and 24-locus mycobacterial interspersed repetitive unit-variable-number tandem repeat (MIRU-VNTR), are used worldwide in studies of Mycobacterium tuberculosis (MTB). Conversely, because of its poor resolution, pulsed-field gel electrophoresis (PFGE) is not widely used for MTB. In this study, we improved the 24-locus MIRU-VNTR and PFGE protocols and compared the effectiveness of these approaches for the molecular typing of MTB using 75 clinical isolates obtained from a cohort investigation of high-risk populations infected with MTB. The 24-locus MIRU-VNTR method demonstrated superior discriminatory ability, followed by PFGE and IS6110-RFLP. Next, we analyzed six isolates with clear epidemiologic connections; that is, isolates from patients who attended the same school. IS6110-RFLP and PFGE identified these samples as the same type. By contrast, according to MIRU-VNTR, two isolates differed from four other isolates at one locus each; one isolate was identified as Mtub29 and the other as QUB-26. In summary, the 24-locus MIRU-VNTR assay was the most useful molecular typing method among the three methods investigated due to its discriminatory power, short time required, and availability as an epidemiologic investigation tool. PFGE was the second-best method. Compared with the other loci assessed in the 24-locus MIRU-VNTR assay, the Mtub29 and QUB-26 loci appeared to exhibit greater variability during transmission.

Molecular Epidemiology of Legionella pneumophila Isolated from Water Supply Systems in Seoul, Korea (서울시 수계시설에서 분리된 Legionella pneumophila의 분자역학적 특성)

  • Jeon, Su Jin;Jung, Ji Hun;Seung, Hyun Jung;Kim, Chang Kyu;Jin, Young Hee;Oh, Young Hee;Choi, Sung Min;Chae, Young Zoo
    • Journal of Environmental Health Sciences
    • /
    • v.39 no.2
    • /
    • pp.166-177
    • /
    • 2013
  • Objectives: The genus Legionella is common in aquatic environments. Some species of Legionella are recognized as potential opportunistic pathogens for human, notably Legionella pneumophila that causes, Legionellosis. Thus, we investigated the contamination of Legionella pneumophila on water supply systems in Seoul, including cooling towers, public baths, hospitals and fountains. Methods: The existence of 16S rRNA and mip gene of L. pneumophila was confirmed in the genome of the isolated strains by PCR. Results: During the summer season of 2010 and 2011, Legionella pneumophila were detected from 163 samples (21.1%) out of 772 samples collected. Among the 163 strains of L. pneumophila, eighty one isolates belonged to serogroup 1 (57.4%), 23 isolates were serogroup 5 (16.3%), 21 isolates were serogroup 6 (14.9%), 8 isolates were serogroup 2 (5.79%), and 8 isolates were identified in serogroup 3 (5.7%). Through PFGE (pulsed-field gel electrophoresis) analysis using Sfi I, genetic types of L. pneumophila were classified into five (A to E) patterns by the band similarity with excess of 70% from public baths. Conclusions: The PFGE patterns of the serotypes showed a tendency for diversity of L. pneumophila. Our results suggest the existence of serological and genetic diversity among the L. pneumophila isolates.