• Title/Summary/Keyword: PFGE

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Serotypes, antimicrobial resistance of Salmonella spp. and plasmid profiles, phage types, PFGE of S. Enteritidis and S. Typhimurium isolated from ducks in Daegu-Gyeongbuk province (대구.경북지역 오리 유래 Salmonella속 균의 혈청형, 항균제 내성 및 S. Enteritidis와 S. Typhimurium의 plasmid profiles, phage types 및 PFGE)

  • Cho, Jae-Keun;Kang, Min-Su;Kim, Ki-Seuk
    • Korean Journal of Veterinary Service
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    • v.34 no.3
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    • pp.217-226
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    • 2011
  • Salmonella spp. is of increasing public health concern as causative pathogens of food poisoning. The aim of this study was to investigate the serotypes and antimicrobial resistance pattern of Salmonella spp. isolated from duck farms in Daegu-Gyeongbuk province. Also, S. Enteritidis and S. Typhimurium isolates were further examined for plasmid analysis, phage typing and pulsed-field gel electrophoresis (PFGE). A total of 34 Salmonella spp. (16.4%) were isolated from duck farms and ten serotypes were identified. The predominant serotypes were S. Typhimurium (23.5%) S. Fyris (17.6%) and S. Haardt (11.8%), S. Agona and S. Enteritidis (respectively 8.8%). Of 34 Salmonella isolates, 15 (44.1%) isolates were resistant to at least one antimicrobial agent and multiple resistance (resistance to more than 4 drugs) was observed in 9 strains (26.5%). The high resistance was found to streptomycin (32.4%), tetracycline (29.4%), ampicillin, kanamycin and nalidixic acid (respectively, 26.5%), all Salmonella isolates were susceptible to cefoxitin, cefotaxime, gentamicin, amikacin and ciprofloxacin. All S. Enteritidis and S. Typhimurium isolates were found to contain only one plasmid (ca. 54 or 55kb, respectively). Among the S. Enteritidis isolates, two phage types were found, PT32a and PT1c, respectively, one isolates did not react with any of the phages used. Whereas, all S. Typhimurium isolates were RDNC (reacts but does not conform). PFGE showed to be a useful typing method better than plasmid analysis and phage typing for discrimination of isolates especially, S. Typhimurium isolates. Our results indicated that the serotypes of Salmonella isolates are widely distributed in duck farms, further epidemiological studies should be carried out.

Comparison of Biotyping, Serotyping and Molecular Typing of Yersinia enterocolitica Isolated from Spring water in Seoul (서울시내 약수에서 분리한 Yersinia enterocolitica의 생물형, 혈청형 및 분자학적 형별비교)

  • 이영기;최성민;오수경;신재영
    • Journal of environmental and Sanitary engineering
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    • v.14 no.4
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    • pp.99-109
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    • 1999
  • Enteropathogenic Yersina enterocolitica is an important cause of human and animal disease. Phenotypic and genotypic characteristics currently used to identify Yersinia enterocolitica are not necessarily sufficient to differentiate pathogenic from non-pathogenic strains or to analyze the epidemiology of yersiniae at a molecular level. To improve the characterization of Yersinia enterocolitica, A total of 65 isolates of Yersinia enterocolitica were examined with bioserotyping, antibiotic susceptibilities, PFGE, PCR-ribotyping. Genomic DNA pattern generated by PFGE are highly specific for different strains of an organism and have significant value in epidemiologic investigations. The PFGE analysis of Not I-digested chromosomal DNA of Y. enterocolitica were performed with a CHEF Mapper(Bio-Rad, USA). Not I generated 19 restriction endonuclease digestion profiles(REDP). PCR-ribotyping, performed with primers complementry to conserved regions of 16S and 23S rRNA gene, generated 13 ribotypes. PCR-ribotyping can be considered a good technich for subtyping strains of Y.enterocolitica.

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Characterization of Multidrug-resistant Salmonella enterica Serovar Typhimurium Isolated from Swine Sources

  • Suh Dong Kyun;Song Jae Chan
    • Biomedical Science Letters
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    • v.11 no.2
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    • pp.115-119
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    • 2005
  • A total of 28 Salmonella enterica serovar Typhimurium isolated from diseased pigs and swine carcasses between 2001 and 2003 were characterized by the antimicrobial resistance profiles, PCR for detection of S. Typhimurium DT104 and pulsed-field gel electrophoresis (PFGE) with the restriction enzyme XbaI. All but one isolate presented multidrug resistance (MDR) to more than two antibiotics tested. A total of 11 resistance profiles were observed, and two phenotypes, ST and ASSuTG, were the most common among them. Two isolates were found to be S. Typhimurium DT104 isolates by PCR, and their resistance profile did not show the DT104 typical resistance type ACSSuT, but ACSSuTGK instead. PFGE identified 11 banding patterns in dendrogram, and three main clusters (designated A to C) were represented. Interestingly, sixteen of 19S. Typhimurium isolates belonging to cluster B showed an identical band pattern.

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Study of Electrophoretic Karyotypes of Fusarium Section Liseola (Fusarium 균의 section Liseola에 대한 핵형 연구)

  • 밍병례;안미선;최영길
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.192-196
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    • 1999
  • CHEF-PFGE(Contour-Clarnped Homogeneous Electric field- Pulsed Field Gel Electrophoresis) was used to identify electrophoretic karyotype for eight strains belonging to the Fzisoriuni section Liseolo. Chromosome numbers were nine to thirteen bands, ranging in size Cram 0.75 to 6.45 Mb. The total genome size was eslimated to range from 38.19 Mb to 43.12 Mb and numerous chromosome-length polymorphisms (CLPs) were observed. For the chromosome localizalion of the gene, 1GS sequence(2.6 Kb) of rDNA from F: moniliforme, chs-2 gene(2.8 Kb) and 4 - 3 gene(3.8 Kb) from Neuuospora cmssa were wed as probes.

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Genetic analysis of Campylobacter jejuni isolates from diarrhea patients in Gyeonggi-do (경기도에서 분리된 Campylobacter jejuni의 유전자 패턴 분석 연구)

  • Kim, Woon-Ho;Choi, Ok-Kyung;Jeong, Jin-A;Park, Sung-Hee;Lee, Yea-Eun;Park, Gwang-Hee;Yoon, Mi-Hye
    • Korean Journal of Microbiology
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    • v.54 no.1
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    • pp.31-37
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    • 2018
  • Campylobacter jejuni is an important food-borne pathogen causing gastroenteritis in human. We isolated 208 strains of Campylobacter jejuni from 430 diarrhea patients and food employees with 17 food-poisoning outbreaks between 2014 and 2016 in Gyeonggi area. The strains were tested for genetic relationship and the genotype distribution using PFGE and multiplex-PCR typing. Among the 47 Penner-serotypes known for C. jejuni, it was identified as a genotype consisting of 35 genotypes by multiplex-PCR typing and represented 7 genotypes (HS2, HS4A, HS8, HS15, HS29, HS41, and HS53) in the selected strain. From the PFGE analysis of 11 food-poisoning outbreaks, 5 group of PFGE profile were obtained, and genetic similarity in these clusters ranged from 61.8 to 66.6%. This study examines the genetic diversity of C. jejuni that have been separated in the Gyeonggi area through various genetic analysis methods and identifies the correlation between strains in patients who have been infected with the disease in the future.

Characterization of Drug-Resistant Salmonella enterica Serotype Typhimurium by Antibiograms, Plasmids, Integrons, Resistance Genes, and PFGE

  • Benacer, Douadi;Thong, Kwai Lin;Watanabe, Haruo;Puthucheary, Savithri Devi
    • Journal of Microbiology and Biotechnology
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    • v.20 no.6
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    • pp.1042-1052
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    • 2010
  • Forty-seven Salmonella Typhimurium (33 zoonotic, 14 clinical) strains were tested for antimicrobial resistance using the standard disk diffusion method. The presence of relevant resistance genes and class 1 integrons were investigated by using PCR. Pulsed-field gel electrophoresis (PFGE) and plasmid profiling were carried out to determine the genomic diversity of Salmonella Typhimurium. Approximately 57.4% of the S. Typhimurium strains were multidrug resistant (MDR) and showed high resistance rates to tetracycline (70.2%), sulfonamides (57.4%), streptomycin (53.1%), ampicillin (29.7%), nalidixic acid (27.6%), kanamycin (23.4%), chloramphenicol (21.2%), and trimethoprim (19.1%). Resistance towards cephalosporins was noted for cephalothin (27.6%), cephradine (21.2%), amoxicillin clavulanic acid (17.0%), and cephalexin (17.0%). Resistance genes, $bla_{TEM}$, strA, aadA, sul1, sul2, tetA, tetB, and tetC, were detected among the drug-resistant strains. Thirtythree strains (70.2%) carried class 1 integrons, which were grouped in 9 different profiles. DNA sequencing identified sat, aadA, pse-1, and dfrA genes in variable regions on class 1 integrons. Thirty-five strains (74.4%) were subtyped to 22 different plasmid profiles, each with 1-6 plasmids (2.0 to 95 kb). PFGE subtyped the 47 strains into 39 profiles. In conclusion, high rates of multidrug resistance were found among the Malaysian Salmonella Typhimurium strains. The emergence of multidrug-resistant Salmonella Typhimurium to cephalosporin antibiotics was also observed. The strains were very diverse and no persistent clone was observed. The emergence of MDR Salmonella Typhimurium is a worldwide problem, and this report provides information for the better understanding of the prevalence and epidemiology of MDR S. Typhimurium in Malaysia.

Prevalence and Characteristics of Antimicrobial-Resistant Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus from Retail Meat in Korea

  • Kim, Yong Hoon;Kim, Han Sol;Kim, Seokhwan;Kim, Migyeong;Kwak, Hyo Sun
    • Food Science of Animal Resources
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    • v.40 no.5
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    • pp.758-771
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    • 2020
  • This study was to investigate the prevalence and characteristics of antimicrobial-resistant Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) from 4,264 retail meat samples including beef, pork, and chicken in Korea between 2013 and 2018. A broth microdilution antimicrobial susceptibility testing was performed for S. aureus. Molecular typing by multilocus sequence typing (MLST), spa typing, and pulsed-field gel electrophoresis (PFGE), was performed on mecA-positive S. aureus strain. S. aureus was isolated at a rate of 18.2% (777/4,264), of which MRSA comprised 0.7% (29 strains). MLST analysis showed that 11 out of the 29 MRSA isolates were predominantly sequence type (ST) 398 (37.9%). In addition, ST72, ST692, ST188, ST9, and ST630 were identified in the MRSA isolates. The spa typing results were classified into 11 types and showed a high correlation with MLST. The antimicrobial resistance assays revealed that MRSA showed 100% resistance to cefoxitin and penicillin. In addition, resistance to tetracycline (62.1%), clindamycin (55.2%), and erythromycin (55.2%) was relatively high; 27 of the 29 MRSA isolates exhibited multidrug resistance. PFGE analysis of the 18 strains excluding the 11 ST398 strains exhibited a maximum of 100% homology and a minimum of 64.0% homology. Among these, three pairs of isolates showed 100% homology in PFGE; these results were consistent with the MLST and spa typing results. Identification of MRSA at the final consumption stage has potential risks, suggesting that continuous monitoring of retail meat products is required.

Comparative Analysis of Salmonella enterica subsp. enterica Serovar Thompson Isolates associated with Outbreaks Using PFGE and wgMLST

  • Youngho Koh;Yunyoung Bae;Min-Jung Lee;Yu-Si Lee;Dong-Hyun Kang;Soon Han Kim
    • Journal of Microbiology and Biotechnology
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    • v.32 no.12
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    • pp.1605-1614
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    • 2022
  • The strains associated with foodborne Salmonella enterica Thompson outbreaks in Korea have not been identified. Therefore, we characterized S. Thompson strains isolated from chocolate cakes linked to foodborne outbreaks in Korea. A total of 56 strains were isolated from preserved cake products, products in the supply chain distribution, the manufacturer's apparatus, and egg white liquid products used for cream preparation. Subsequently, serological typing, pathogenic gene-targeted polymerase chain reaction (PCR), pulsed-field gel electrophoresis (PFGE), and whole-genome multi-locus sequence typing (wgMLST) were performed to characterize these isolates. The antigen formula of all isolates was 7:k:1,5, namely Salmonella enterica subsp. enterica Serovar Thompson. All 56 isolates harbored invA, his, hin, and stn, and were negative for sefA and spvC based on gene-targeted PCR analyses. Based on PFGE results, these isolates were classified into one group based on the same SP6X01.011 pattern with 100% similarity. We selected 19 strains based on the region and sample type, which were subjected to wgMLST. Although the examined strains showed 100% similarity, they were classified into seven clusters based on allelic differences. According to our findings, the cause of these outbreaks was chocolate cake manufactured with egg white liquid contaminated with the same Salmonella Thompson. Additionally, comparative analysis of wgMLST on domestic isolates of S. Thompson from the three outbreaks showed genetic similarities of over 99.6%. Based on the results, the PFGE and wgMLST combination can provide highly resolved phylogeny and reliable evidence during Salmonella outbreak investigations.

Pulsed-Field Gel Electrophoresis and Monoclonal Antibody Analysis of Leptospira interrogans Isolated in Korea (국내 분리 렙토스피라균의 단클론 항체 및 Genomic DNA의 Pulsed-Field Gel Electrophoresis 분석)

  • 조민기;기선호;김형준;김윤원;장우현;오희복
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.197-204
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    • 1999
  • A total of 22 Leptospiua inlermgans field isolates from the ~ a t s captured in 5 provinces of Korea in 1996, and 6 antigenically closely related relerence serovars of lai, yeonchon, birkini. gem, mwogolo. and canicola were analysed. When the antigenic characteristics were analysed by reactivity with 7 monoclonal antibodies prepared with sh.ains belongng to serogroup Icterohaemorrhagiae. all 22 isolates showed the same reaction pattern with that of serovar lai. Large restriction fragment patterns obtained after cleavage of geno~nic DNAs with infrequently cuttimg restriction enzymes were analyzed by pulsed-field pel electrophoresis(PFGE). Identification of leptospira strains by PFGE with Nor I, Asc I or Iise I digests correlated with their antigenically typed serovars, silh a few exceptions. PFGE of isolates, except for JR89, digested wjth Nor I showed identical pattern w~th serovar lai, showing 13 Cragments between 940 kb and 63 kb. When PFGE pallerns of JR89 were compared with those of serovar lai, Not I digest showed additional two hands of 1000 kb and 460 kb, while Asc I digest showed 650 kb fragment and Fse I digest did not show the fragment of 280 kb. Whereas serovar yeonchon. which was isolated in Korea and identified as a new serovar previously. could be differentiated from serovar lai in antigenic reactivities with monoclonal antibodres. it showed the similar PFGE pattern with serovar lai includin~ reference and field isolates. It was suggested that Korean leptospiral field isolates are closely related in DNA level.

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Analysis of Epidemiological Characteristics, PFGE Typing and Antibiotic Resistance of Pathogenic Escherichia coli Strains Isolated from Gyeonggi-do (경기도에서 분리한 병원성대장균의 역학적 특성 및 PFGE, 항생제 내성 연구)

  • Kim, Kyung-A;Yong, Kum-Chan;Jeong, Jin-A;Huh, Jeong-Weon;Hur, Eun-Seon;Park, Sung-Hee;Choi, Yun-Sook;Yoon, Mi-Hye;Lee, Jong-Bok
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.285-295
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    • 2014
  • This study was conducted to survey the epidemiological characteristics and the isolated strains for pathogenic E. coli which was the major causative organisms for food poisoning occurred at school food services in the Gyeonggi-do area during the past three years. We investigated 19 accidents of food-borne disease outbreaks by pathogenic E. coli at school food services from 2010 to 2012. Food-borne disease outbreaks by pathogenic E. coli were usually occurred at direct management type (18 accidents, 95%) and high schools. For the seasonal factors, 13 accidents (65%) were occurred in June to September, especially the end of August and September after the summer holidays. The first patients were occurred on Wednesday (7 accidents, 37%) and Thursday (7 accidents, 37%), and they were mainly reported on Thursday (7 accidents, 37%) and Friday (5 accidents, 26%). The exposure of risk was estimated in Monday (4 accidents, 21%), Tuesday (7 accidents, 37%) and Wednesday (4 accidents, 21%), and kimchi (5 accidents, 50%) was estimated as the food of the high risk responsible for the outbreaks. 98 isolates of pathogenic E. coli consisted of PEC (50%), ETEC (34%), EAEC (15%), and EHEC (1%). The antibiotic resistance of pathogenic E. coli showed in the descending order of ampicilline (40%), nalidixic acid (37%), trimethoprim/sulfamethoxazole (24%), and tetracycline (19%). The antibiotics of second and third generation cephalosporins, cabarpenem, aminoglycosides, and second generation quinolones had antimicrobial susceptibilities and cefalotin, ampicillin/sulbactam and chloramphenicol showed medium resistance at 29%, 25%, and 6% respectively, and 70% of isolates were resistant to more than one antibiotic. By the PFGE analysis, they were classified into nine major groups and 31 profiles with 57% pattern similarity. It was very difficult to find the correlation of antimicrobial susceptibilities and genotype in the small scale-food poisoning, but the similarity of antimicrobial resistance and PFGE patterns in the large scale-food poisoning enabled the outbreaks to estimate the same pathotype of E. coli derived from identical origins.