• Title/Summary/Keyword: PFGE patterns

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Antimicrobial resistance and pulsed -field gel electrophoresis patterns of Salmonella gallinarum isolated from broiler (육계에서 분리한 Salmonella gallinarum 의 약제내생 및 PFGE 양상)

  • Kim Seong-Guk;Kim Yeong-Hwan;Eom Hyun-Jung;Jang Seong-Jun;Jo Gwang-Hyeon;Lee Yang-Soo
    • Korean Journal of Veterinary Service
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    • v.29 no.3
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    • pp.297-308
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    • 2006
  • Fowl typhoid (FT) is a septicemic disease caused by Salmonella gallinarum. The purpose of this study was to investigate the antimicrobial resistance and pulsed -field gel electrophoresis (PFGE) patterns of S gallinarum isolated from broiler. During 1999 to 2004, there was isolated a total of 26 strains in liver and spleen. The biochemical characteristics of S gallinarum isolates was nonmotile, no production of $H_2S$, glucose gas, non-fermented rhamnose, indole-negative, fermentation of dulcitol, mannitol, maltose, and ornithine decarboxylase. At antimicrobial susceptibility, all of isolates were susceptible to amoxicillin/clavulanic acid, amikacin, neomycin, kanamycin, and cephalothin. Twenty-six isolates were divided into 19 resistant patterns and 6 strains was 8-multi-drug resistance. PFGE of Xba I restriction fragments of S gallinarum isolates was 22 patterns.

Genomic analysis of Mycobacterium fortuitum by pulsed-field gel electrophoresis (Pulsed-field Gel Electrophoresis를 이용한 Mycobacterium fortuitum의 유전형 분석)

  • Lee, Tae-Yoon;Do, In-A;Kim, Sung-Kwang
    • Journal of Yeungnam Medical Science
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    • v.12 no.2
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    • pp.366-385
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    • 1995
  • Epidemiological studies are important in both the prevention and treatment of mycobacterial infections. This study was initiated to establish the pulsed-field gel electrophoresis (PFGE) method, which are not yet extensively studied. The most apprpriate restriction endonucleases included DraI, AsnI, and XbaI. The optimal PFGE condition was different according to the enzymes used. Two stage PFGE was performed, in case of DraI first stage was performed with 10 seconds of initial pulse and 15 seconds of final pulse, while the second stage was performed with 60 seconds of initial pulse and 70 seconds of final pulse. The electrophoresis time for DraI-PFGE was 14 hours for each stage. Electrophoresis was performed for 22 hours, in case of XbaI, with 3 seconds of initial pulse and 12 seconds of final pulse. Electrophoresis was performed for 22 hours, in case of AsnI, with 5 seconds of initial pulse and 25 seconds of final pulse. In all cases the voltage of the electrophoresis was maintained constantly at 200 voltage. Standard mycobacterial strains, which included Mycobacterium bovis BCG, M. tuberculosis, and M. fortuitum, could not be differentiated by PFGE analysis. PFGE analysis was performed to differentiate 9 clinically isolated M. fortuitum strains using AsnI. All M. fortuitum strains showed different genotypes except 2 strains. Cluster analysis divided M. fortuitum strains into 2 large groups. PFGE analysis was performed to further differentiate M. fortuitum isolates using XbaI. The undifferentiated 2 M. fortuitum strains showed different PFGE patterns with Xba I. Cluster analysis of the XbaI-PFGE patterns showed more complex grouping than AsnI-PFGE patterns, which showed that XbaI-PFGE analysis was better than AsnI-PFGE in M. fortuitum genotyping. The top dissimilarity values of AsnI-PFGE and XbaI-PFGE were 0.74 and 0.75, respectively. This value was higher than that of arbitrarily primed polymerase chain reaction (AP-PCR) analysis and lower than that of restriction fragment length polymorphism (RFLP) analysis. This suggested that PFGE can be used as a supportive or alternative genotyping method to RFLP analysis.

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Discrimination of Listeria monocytogenes by Sequence Typing Based on Two Housekeeping Genes and Its Comparison to PFGE Patterns

  • Suh, Dong-Kyun
    • Biomedical Science Letters
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    • v.11 no.3
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    • pp.289-293
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    • 2005
  • Two housekeeping genes, of Listeria monocytogenes dat and hlyA, were analyzed in a set of 28 isolates from different sources to estimate their genetic diversities. These strains were previously characterized by pulsed-field gel electrophoresis. Complete gene sequences for dat (465 bp) and hlyA (584 bp) had sequence similarity of $99.87-100\%$ S and $99.96-100\%$ S among isolates, respectively. Also, we found that the numbers of sequence types (ST) were about 3-fold less than those of PFGE types (3 STs versus 11 PFGE types). There was, however, a good correlation between the PFGE patterns and phylogenetic grouping of two gene sequences among the isolates. Further studies on analyzing additional loci would increase the discriminatory power of sequence typing for L. monocytogenes strains.

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Epidemiological analysis of Escherichia coli O157 : H7 by pulsed-field gel electrophoresis and multiplex polymerase chain reaction

  • Jung, Byeong-yeal;Jung, Suk-chan;Cho, Dong-hee;Kim, Jong-yeom;Kim, Bong-hwan
    • Korean Journal of Veterinary Research
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    • v.39 no.2
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    • pp.338-342
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    • 1999
  • Twenty three strains of Escherichia (E) coli O157 : H7 isolated from Korea, Japan, USA were analyzed by pulsed-field gel electrophoresis (PFGE) of XbaI-digested chromosomal DNA and multiplex polymerase chain reaction. Various PFGE patterns of E. coli O157 : H7 were found on the same farm. Most of the E, coli O157 : H7 strains had shiga-like toxin (slt) II gene only (43.5%) or both slt I and slt II genes(30.4%). eaeA gene was highly conserved in the E. coli O157 : H7. There was no correlation between PFGE and slt gene patterns. The results indicate that various genotypes of E. coli O157 : H7 have spread throughout the country and genomic DNA patterns generated by PFGE are highly specific for different strains and have significant value in epidemiologic investigations of infectious disease outbreaks.

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Phage typing and pulsed-field gel electrophoresis of Salmonella typhimurium and S enteritidis isolated from domestic animals in Gyeongbuk province (경북지역 가축에서 분리된 Salmonella typhimurium과 S enteritidis의 phage typing 및 pulsed-field gel electrophoresis)

  • Kim, Sang-Yun;Lee, Hee-Moo;Kim, Sin;Hong, Hyon-Pyo;Kwon, Heon-Il
    • Korean Journal of Veterinary Service
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    • v.24 no.3
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    • pp.243-253
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    • 2001
  • Forty-five Salmonella typhimurium isolates were encountered 8 phage types in which DT197 and U302 were the predominant types. The DT104 type which was first found from pig in Korea, and was resistant to chloramphenicol, streptomycin, sulfamethoxazole/trimethoprim, tetracycline, gentamicin and nalidixic acid. Twenty-two S enteritidis isolates were encountered 5 phage types in which PT4 were the representative (predominant). S enteritidis isolates were susceptible to all antimicrobial agents. As a result of PFGE analysis for S typhimurium and S enteritidis, PFGE patterns was better than phage typing in discriminating of strains. PFGE patterns were not in accord with phage type even though some strain had the same phage types.

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Molecular Typing of Salmonella enterica serovar Typhi Strains Isolated in Busan by Pulsed-Field Gel Electrophoresis (부산지역에서 분리된 Salmonella enterica serovar Typhi균에 대한 PFGE를 이용한 Molecular typing)

  • Min, Sang-Kee;Lee, Ju-Hyun;Park, Eun-Hee;Kim, Jung-Ah;Kim, Kyu-Won
    • Journal of Life Science
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    • v.16 no.4
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    • pp.664-671
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    • 2006
  • We obtained 424 Salmonella enterica serovar Typhi isolates from sporadic cases of infection in Busan during 1996 to 2005. We investigated the trend of antimicrobial resistance and molecular typing by pulsed-field gel electrophoresis (PFGE). Of the total 424 isolates, 6 strains (1.4%) were multidrug-resistant (MDR) S. enterica serovar Typhi isolates, 2 strains (0.5%) were resistant to only nalidixic acid, and the remaining 416 strains (98.1%) were fully susceptible to the 18 antimicrobial agent. PFGE of XbaI-digested chromosomal DNA was performed on 50 sporadic S. enterica serovar Typhi isolates with the objective of investigating the extent of genetic diversity of these isolates in our region. We could find that these isolates were much more heterogeneous and at least 32 different PFGE patterns were generated according by dice coefficient, between 0.69 and 1.0. Restriction fragment patterns consisted of 13 to 18 fragments ranged in size from 20 to 630 kb. The results confirmed that PFGE would be an useful tool for investigating surveillance of sporadic or outbreak case and assessing clonality for S. enterica serovar Typhi in Busan area. Our finding will be valuable in developing rational strategies to control this pathogen and setting the basis of an effective PulseNet system in Korea.

Analysis of antimicrobial resistance and PFGE patterns of Salmonella spp. isolated from chickens at slaughterhouse in Incheon area (인천지역 닭 도축장에서 분리된 Salmonella spp.의 항생제 내성 및 PFGE 패턴분석)

  • Yang, Ha-Young;Lee, Sung-Mo;Park, Eun-Jeong;Kim, Jung-Hee;Lee, Jung-Goo
    • Korean Journal of Veterinary Service
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    • v.32 no.4
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    • pp.325-334
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    • 2009
  • Salmonella spp. are the important pathogens both economically and clinically in animals as well as human. Some of them have highly zoonotic potentials even though they are asymptomatic in animals. Therefore, the prevalence of Salmonella spp. in animals is highly concerned for human health. The present study was carried out to investigate the prevalence, antimicrobial resistance and PFGE patterns of Salmonella spp. isolated from chickens at slaughterhouse in Incheon area. The overall isolation rate of Salmonella spp. from cloaca and cecum specimens was 7.3 % (37/510). Thirty seven isolates of Salmonella spp. were identified to 5 serotypes; S. Enteritidis, S. Newport, S. Typhimurium, S. Gallinarum, and S. Derby with prevalence of 46.0%, 40.5%, 8.1%, 2.7%, and 2.7%, respectively. Resistance to nalidixic acid was found in 97.3% of Salmonella spp. isolated, followed by streptomycin (16.2%), tetracycline (16.2%), ampicillin (5.4%). Only 6 isolates (16.2%) showed resistance to more than two antimicrobials. In PFGE analysis of chicken and human isolates with Xba I, S. Enteritidis isolates from chicken showed very high similarity over 82.8% and also the similarity was very high in the comparison with human isolates. However, the higher similarity (100%) was observed among chicken isolates of S. Typhimurium. These results suggest the close genetic relatedness of Salmonella spp. isolated from chickens with human.

Characterization of Antimicrobial Resistance Patterns and Integrons of Nontyphoid Salmonella Isolates from Infants in Seoul (서울지역 소아에서 분리된 Nontyphoid Salmonella의 항생제 내성과 Integron의 특징)

  • Jin, Young-Hee;Kim, Jin-Ah;Jung, Ji-Hun;Jeon, Soo-Jin;Lee, Jae-Kyoo;Oh, Young-Hee;Han, Ki-Young;Lee, Young-Ki
    • Korean Journal of Microbiology
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    • v.46 no.4
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    • pp.326-333
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    • 2010
  • A total of 105 nontyphoid Salmonella isolated from infants in Seoul from 2003 to 2009 was investigated for their serotype, antimicrobial resistance, characterization of integron, and the patterns of Pulsed-field gel electrophoresis (PFGE). Eighteen serotypes were detected in 105 isolates, and the two most common serotypes were S. Enteritidis (47.6%) and Montevideo (15.2%). Among the Salmonella serovars, a high level of antimicrobial resistance was found to ampicilin (60%), tetracycline (46.7%), streptomycin (35.2%) and nalidixic acid (28.6%). In the multi-drug resistance patterns, the predominant patterns were only nalidixic acid (15.7%), ampicillin-ampicillin/sulbactam-tetracycline (14.5%), and ampicillin-streptomycin-chloramphenicol-tetracycline (10.8%). PCR and DNA sequencing analysis revealed the presence of class 1 integron in 20 isolates (19%). Of the class 1 integron positive isolates 20% harboured the integron-associated gene cassettes : aadA2, blaP1, dfr17-aadA5, dfrA12-aadA2, and aadA7. PFGE was carried out to examine the genetic relatedness among S. Enteritidis isolates. Except for three strains, fifty strains were divided by three pulsotypes.

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.

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.