• The Journal of the Korean Society for Microbiology
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• v.35 no.4
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• pp.289-297
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• 2000

Background: Staphylococcus aureus (s. aureus) and Escherichia coli (E. coli) are major pathogens in community and hospital. And they sometimes cause the outbreak in hospital in the immunocompromised patients. Pulsed-field gel electrophoresis (PFGE) has been regarded as a standard method for genotyping in epidemiologic studies, but it is laborious and time-consuming. Infrequent restriction site-polymerase chain reaction (IRS-PCR), a new genotyping methods, was performed to compare the applicability with PFGE. Methods: We performed PFGE and IRS-PCR on S. aurues (n=120) and E. coli (n=117) which were collected clinically in 4 different hospitals. We assessed each method in terms of discriminatory power, quality, and efficiency. Results: In E. coli, the discriminatory power of IRS-PCR was $46.7{\sim}86.7%$, and that of PFGE was $88.9{\sim}96.7%$ according to hospital. But in S. aurues, the discriminatory power of IRS-PCR was $20{\sim}56.7%$, and that of PFGE was $40{\sim}90%$ according to hospital. The typablity and reproducibility of IRS-PCR were 100% of each. PFGE needed four days to complete the procedure, but IRS-PCR could be performed within one day, IRS-PCR showed better resolution than PFGE. Conclusion: In case of gram negative bacteria (like E. coli), IRS-PCR could be a reliable alternative for epidemiologic typing due to better efficiency and comparable discriminatory power. But in the case of gram positive bacteria (like S. aureus), IRS-PCR does not seem to be suitable for the strain-to-strain differentiation. More trials and changes of restriction enzymes or primers could reveal the efficacy of IRS-PCR in the field of molecular typing.

• 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.

• 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.

• 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.

• Korean Journal of Veterinary Service
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• v.30 no.3
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• pp.363-374
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• 2007

A biochemical characterization and antimicrobial drugs susceptibility study was conducted in four breeding kennel which was canine abortion caused by Brucella canis in Gyeongbuk province in 2003-2006. Total of 267 dogs domesticated in the four kennel were examination. Among them, 143 (53.6%) dogs were sero-positive and 25 of blood samples were isolated to Brucella canis. At amplification of 35KDa-BCSP gene using PCR, 711 bp DNA fragment was same visible in 25 isolates and B canis RM6/66. Biochemical characterization of B canis isolated was non-hemolytic, no production of $H_2S$, no fermentation of carbohydrates, catalase-positive, oxidase-positive, indol-negative, hydrolyzation of urea, reduction of nitrate and development of thionin dye medium. Using disk-diffusion method, all of 25 strains tested were found to be highly susceptible to tetracycline, aminoglycoside, quinolone, macrolide antibiotics, rifampin and ampicillin in vitro. Using PFGE with restriction enzyme Smi I, 25 isolates tested were typed to 2 pattern, S1 and S2.

• Journal of Microbiology
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• v.44 no.3
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• pp.336-343
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• 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.

• Journal of Microbiology
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• v.43 no.5
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• pp.424-429
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• 2005

In previous studies, it has been reported that both S. enteritidis, the most common serotype, and S. enteritidis Phage Type 4 (SEPT 4) isolates were identified as the most prevalent PT in domestic poultry and also in humans in Korea until 2002. The aim of this study was to analyze the genetic diversity and epidemiological properties of both PT isolates, and also to trace the source of SEPT 4 isolates from domestic poultry and humans by Pulsed-field gel electrophoresis (PFGE). In order to understand the molecular epidemiologic properties of SEPT 4 isolates, which have very similar phenotypic properties to our preliminary investigations (serotyping, phage typing, large plasmids and antibiograms), PFGE analysis with XbaI enzyme was performed on the representative SEPT 4 isolates. Thirty-six SEPT 4 isolates were analyzed and differentiated with 10 pulsed-field profiles (PFP) expressing very high discriminative ability (SID: 0.921). In PFP, SEPT 4 isolates from human patients showed a perfect genetic match with those from broiler chickens and meats. Therefore, this study was able to successfully trace the major source of SEPT 4 isolates and also to determine the usefulness of the PFGE method for genetic analysis of epidemic strains.

• 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.

• The Journal of the Korean Society for Microbiology
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• v.35 no.3
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• pp.239-250
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• 2000

Sixty-eight clinical isolates of Stenotrophomonas maltophilia from inpatients of 2 university hospitals in Taegu were epidemiologically analyzed by using the minimum inhibitory concentrations of 25 antimicrobial drugs, biochemical reaction, pulsed-field gel elctropgoresis (PFGE), and PCR with enterobacterial repetitive intergenic consensus sequences as primer (ERIC-PCR). 1. All the strains were susceptible to minocycline. More than 57% were susceptible to sulfisomidine (Su), ciprofloxacin (Ci), Ofloploxacin (Of), nalidixic acid (Na), and chloramphenicol (Cm), and $19{\sim}35%$ to ceftazidime (Cd), trimethoprim (Tp), Ticacillin-clavulanic acid, and cefoperazone-sulbactam. Most isolates were resistant to ${\beta}$-lactam antibiotics such as ampicillin (Ap), carbenicillin (Cb), cefotaxim (Ct), cefoxitin (Cx), and aminoglycosides including gentamicin (Gm), tobramycin (Tb), amikacin (Ak). 2. All the isolates were multiply resistant of 5 to 17 drugs and showed 40 different resistance pattern types. 3. All the strains showed very similar biochemical reactions except ${\beta}$-galactosidase and nitrate reduction test. Fourteen strains selected randomly were classified 10 different pattern type by PFGE and ERIC-PCR. These two methods showed identical result. Four strains isolated from wound in 1994 showed similar MIC pattern and identical API 20NE profile, PFGE, and ERIC-PCR pattern indicating episodes of cross-infection among patients. These results indicate that PFGE or ERIC-PCR profile has comparable discriminatory power for epidemiological typing of S. maltophilia.

• Korean Journal of Veterinary Service
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• v.30 no.3
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• pp.353-362
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• 2007

A total of 1,354 samples was collected from bovine and porcine carcass from January 2005 to December 2006 in a slaughter house. Twenty five strains(1.8%) of Listeria monocytogenes were isolated from 1,354 samples using selective media. Ten(1.4%) L monocytogenes were isolated from the 677 of bovine carcasses, and 15(2.2%) were isolated from the 677 of porcine carcasses. Among 15 L mono-cytogenes from porcine, 11 siolates were serovars 1/2c, followed by 1/2b (3 strains, 20.0%) and 1/2a(1 strain) Out of 10 bovine samples, positive cases in 1/2a were 9 strains (90.0%), 1/2b were 1 strains(10.0%). PCR primers were selected to amplify a 520-base pair(bp) DNA fragment from the listeolysin O gene (hlyA) of L mono-cytogenes. A 520-bp product was detected in PCR with DNA from L monocytogenes, but not from the other Listeria species tested. A total of 25 L monocytogenes strains were analysed by PFGE after digestion with Apa I. PFGE analysis of genomic DNA showed the $14{\sim}18$ fragments ranging in size from 30 to 550 kb, resulting in 14 patterns.