• 대한수의학회지
• /
• 제54권2호
• /
• pp.91-99
• /
• 2014

An antimicrobial susceptibility test was conducted to compare the resistance rates among Campylobacter spp. isolates from dogs (n = 50) raised under diverse conditions and humans (n = 50). More than 60% of Campylobacter (C.) jejuni from dogs and humans showed resistance to nalidixic acid, enrofloxacin and ciprofloxacin. C. jejuni isolates from humans showed higher resistance to tetracycline (83.3%) and ampicillin (91.3%) than those from dogs. None of the C. jejuni or Campylobacter coli isolates from humans or dogs were resistant to erythromycin. Overall, 85% of Campylobacter spp. isolates showed a multidrug resistant phenotype. Nucleotide sequencing analysis of the gryA gene showed that 100% of $NA^R/CIP^R$ C. jejuni isolates from dogs and humans had the Thr-$86^{th}$-Ile mutation, which is associated with fluoroquinolone resistance. flaA PCR restriction fragment length polymorphism (RFLP) typing to differentiate the isolates below the species level revealed 12 different clusters out of 73 strains. The human isolates belonged to eight different RFLP clusters, while five clusters contained dog and human isolates.

• Journal of Microbiology and Biotechnology
• /
• 제21권12호
• /
• pp.1336-1344
• /
• 2011

This study details and examines a novel ligation-mediated polymerase chain reaction (LM-PCR) method. Named the LM-PCR/Shifter, it relies on the use of a Class IIS restriction enzyme giving restriction fragments with different 4-base, 5' overhangs, this being the Shifter, and the ligation of appropriate oligonucleotide adapters. A sequence of 4-base, 5' overhangs of the adapter and a 4-base sequence of the 3' end of the primer(s) determine a subset of the genomic restriction fragments, which are amplified by PCR. The method permits the differentiation of bacterial species strains on the basis of the different DNA band patterns obtained after electrophoresis in polyacrylamide gels stained with ethidium bromide and visualized in UV light. The usefulness of the LM-PCR/Shifter method for genotyping is analyzed by a comparison with the restriction endonuclease analysis of chromosomal DNA by the pulsed-field gel electrophoresis (REA-PFGE) and PCR melting profile (PCR MP) methods for isolates of clinical origin. The clustering of the LM-PCR/Shifter fingerprinting data matched those of the REA-PFGE and PCR MP methods. We found that the LM-PCR/Shifter is rapid, and offers good discriminatory power and excellent reproducibility, making it a method that may be effectively applied in epidemiological studies.

• Journal of Microbiology and Biotechnology
• /
• 제25권7호
• /
• pp.1154-1162
• /
• 2015

The emergence of carbapenem resistance among Pseudomonas aeruginosa is an increasing problem in many parts of the world. In particular, metallo-$\beta$-lactamases (MBLs) and AmpC $\beta$lactamases are responsible for high-level resistance to carbapenem and cephalosporin. We studied the diversity and frequency of $\beta$-lactamases and characterized chromosomal AmpC $\beta$lactamase from carbapenem-resistant P. aeruginosa isolates. Sixty-one carbapenem-resistant P. aeruginosa isolates were collected from patients in a tertiary hospital in Daejeon, Korea, from January 2011 to June 2014. Minimum inhibitory concentrations (MICs) of four antimicrobial agents were determined using the agar-dilution method. Polymerase chain reaction and sequencing were used to identify the various $\beta$-lactamase genes, class 1 integrons, and chromosomally encoded and plasmid-mediated ampC genes. In addition, the epidemiological relationship was investigated by multilocus sequence typing. Among 61 carbapenem-resistant P. aeruginosa isolates, 25 isolates (41.0%) were MBL producers. Additionally, 30 isolates producing PDC (Pseudomonas-derived cephalosporinase)-2 were highly resistant to ceftazidime (MIC₅₀ = $256{\mu}g/ml$) and cefepime (MIC₅₀ = $256{\mu}g/ml$). Of all the PDC variants, 25 isolates harboring MBL genes showed high levels of cephalosporin and carbapenem resistance, whereas 36 isolates that did not harbor MBL genes revealed relatively low-level resistance (ceftazidime, p < 0.001; cefepime, p < 0.001; imipenem, p = 0.003; meropenem, p < 0.001). The coexistence of MBLs and AmpC $\beta$-lactamases suggests that these may be important contributing factors for cephalosporin and carbapenem resistance. Therefore, efficient detection and intervention to control drug resistance are necessary to prevent the emergence of P. aeruginosa possessing this combination of $\beta$-lactamases.

• 대한의생명과학회지
• /
• 제12권3호
• /
• pp.267-274
• /
• 2006

Klebsiella pneumoniae is the leading cause of nasocomial infection and the most commonly isolated from clinical specimens. $Extended-spectrum-{\beta}-lactamase$ (ESBL) producing K. pneumoniae infection was associated with a significantly longer duration of hospital stay and greater hospital charges. The purpose of this study is to investigate the antibiotic resistant patterns and the DNA fingerprint types of extended-spectrum ${\beta}-lactamase$ (ESBL) producing K. pneumoniae. 223K. pneumoniae strains were collected from three general hospitals with more than 500 beds in Busan, Korea from September 2004 to October 2005. The minimum inhibitory concentration (MIC) of antibiotics was measured using the Gram negative susceptibility (GNS) cards of VITEK (Vitek system, Hazelwood Inc., MO). Random amplified polymorphic DNA method was used to detect DNA fingerprint of the organisms. Of the 226 K. pneumoniae isolates 65 ESBL-producing K. pneumoniae strains were detected by the Vitek system and confirmed by the double-disk synergy test. All the 65K. pneumoniae strains were resistant cefazolin, cefepime, ceftriaxone and aztreonam, and 83.0% of the organisms were resistant to ampicillin/sulbactam, 66.1% to tobramycin, 67.6% to piperacillin/tazobactam, 61.5% to ciprofloxacin, and 47.6% to trimethoprim/sulfamethoxazole and 43.0% to gentamicin. The RAPD patterns were distincted as 10 types by three random 10-mer primers (208, 272, 277). Among ten type patterns, three types (Ic, IIb, IIIe) were remarkably represented at patient of internal department, nerve surgery department, general surgery department, and neonatal room. These results indicate that RAPD can be useful for DNA of strains typing in the epidemiological investigations. Therefore more investigation are needed in order to prevent the ESBL type-producing K. pneumoniae from spreading resistance to oxyimino cephalosphorin antibiotics.

• 사상체질의학회지
• /
• 제23권4호
• /
• pp.471-486
• /
• 2011

1. Objectives The purpose of this study was to investigate the high-sensitivity C-reactive protein (hsCRP) related characteristics found in each Sasang Constitutional type. 2. Methods This cross-sectional epidemiological study was conducted using data from 2,842 men and women. After typing each participant into one of the four Sasang constitutional types, they were analyzed on various cardiovascular disease-related variables. 3. Results 1) The predictors of cardiovascular disease (metabolic syndrome, high risk FRS group, high risk hsCRP group) was found to show the highest prevalence in the Taeeum type. 2) In the group with metabolic syndrome, the mean hsCRP concentration in the Taeeum and Soyang types were higher than in the Soeum type. In the group without metabolic syndrome, the mean hsCRP concentration in the Taeeum type were higher than in the Soyang and Soeum types. 3) In the FRS low risk group, the mean hsCRP concentration were higher in order of Taeeum type > Soyang type > Soeum type. In the FRS high risk group, the mean hsCRP concentrations were not significantly different among the Sasang constitutional types. 4. Conclusions The results of this study suggest that the Taeeum constitutional type is a risk factor for high hsCRP and cardiovascular disease. These findings suggest that the prevention of cardiovascular disease is more important in the Taeeum type compared to other constitutional types. Moreover, preventive measures are warranted even in the Taeeum-type persons with low clinical cardiovascular risk.

• 대한미생물학회지
• /
• 제35권3호
• /
• pp.239-250
• /
• 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.

• Clinical and Experimental Pediatrics
• /
• 제60권7호
• /
• pp.221-226
• /
• 2017

Purpose: Escherichia coli sequence type (ST) 131, a multidrug-resistant clone causing extraintestinal infections, has rapidly become prevalent worldwide. However, the epidemiological and clinical features of pediatric infections are poorly understood. We aimed to explore the characteristics of ST131 Escherichia coli isolated from Korean children with urinary tract infections. Methods: We examined 114 uropathogenic E. coli (UPEC) isolates from children hospitalized at Chung-Ang University Hospital between 2011 and 2014. Bacterial strains were classified into STs by partial sequencing of seven housekeeping genes (adk, fumC, gyrB, icd, mdh, purA, and recA). Clinical characteristics and antimicrobial susceptibility were compared between ST131 and non-ST131 UPEC isolates. Results: Sixteen UPEC isolates (14.0%) were extended-spectrum ${\beta}-lactamase$ (ESBL)-producers; 50.0% of ESBL-producers were ST131 isolates. Of all the isolates tested, 13.2% (15 of 114) were classified as ST131. There were no statistically significant associations between ST131 and age, sex, or clinical characteristics, including fever, white blood cell counts in urine and serum, C-reactive protein, radiologic abnormalities, and clinical outcome. However, ST131 isolates showed significantly lower rates of susceptibility to cefazolin (26.7%), cefotaxime (40.0%), cefepime (40.0%), and ciprofloxacin (53.3%) than non-ST131 isolates (65.7%, 91.9%, 92.9%, and 87.9%, respectively; P<0.001 for all). ESBL was more frequently produced in ST131 (53.3%) than in non-ST131 (8.1%) isolates (P<0.01). Conclusion: ST131 E. coli isolates were prevalent uropathogens in children at a single medical center in Korea between 2011 and 2014. Although ST131 isolates showed higher rates of antimicrobial resistance, clinical presentation and outcomes of patients were similar to those of patients infected with non-ST131 isolates.

• Journal of Microbiology and Biotechnology
• /
• 제11권5호
• /
• pp.788-797
• /
• 2001

The usefulness of three PCR methods were evaluated for the epidemiological typing of Staphylococcus aureus: an enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR), repetitive extragenic palindromic element PCR (REP-PCR), and 16S-23S intergenic spacer PCR (ITS-PCR). The analysis was performed using a collection of S. aureus strains comprised of 6 reference and 79 isolates from patients with various diseases. Among the 85 S. aureus strains tested, 6 references and 6 isolates were found to be susceptible to methicillin, whereas the remaining 73 isolates were resistant to it. PCR methods are of special concern, as conventional phenotypic methods are unable to clearly distinguish among methicillin-resistant S. aureus (MRSA) strains. The ability of the techniques to detect different unrelated types was found to be as follows: ERIC-PCR, 19 types; REP-PCR, 36 types; and ITS-PCR, 14 types. On the basis of combining the ERIC, REP, and ITS fingerprints, the 85 S. aureus strains were grouped into 56 genetic types (designated G1 to G56). The diversities for the 85 S. aureus strains, calculated according to Simpson\`s index, were 0.88 for an ERIC-PCR, 0.93 for a REP-PCR, and 0.48 for an ITS-PCR, and the diversity increased up to 0.97 when an ERIC-PCR and REP-PCR were combined. The above discrimination indices imply that the genetic heterogeneity of S. aureus strains is high. Accordingly, this study demonstrates that DNA sequences from highly conserved repeats of a genome, particularly a combination of ERIC sequences and REP elements, are a convenient and accurate tool for the subspecies-specific discrimination and epidemiologic tracking of S. aureus.

• 한국동물위생학회지
• /
• 제35권1호
• /
• pp.69-71
• /
• 2012

Rabbits are highly susceptible to colibacillosis, and no treatment is able to control the disease effectively. Rabbits raised in a farm in Chonbuk province presenting persistent diarrhea and death were submitted for diagnosis. Ninety percent of the infected animals died; weanlings suffered the most mortality. Necropsies showed prominent hemorrhagic foci along the intestinal and cecal serosae. In histopathological examination, rod-shaped bacteria were observed in the necrotic areas of the tips of villi in the small intestine and neutrophils infiltration was found around the necrotic villous areas. The affected animals consistently yielded Escherichia coli isolates from the intestines. The isolated organism was atypically indole-negative and was nonserotypable using 62 known O group-typing sera. Further microbiological and epidemiological works to recognize and control colibacillosis infection in farmed rabbits in Korea is therefore critical.

• Restorative Dentistry and Endodontics
• /
• 제8권1호
• /
• pp.31-44
• /
• 1982

Streptococcus mutans strains were isolated from dental plaques of carious lesions of 53 patients on mitis-salivarius-bacitracin (MSB) and mitis-salivarius(MS) medium as a supplement. The epidemiological investigation was carried out to determine the biotypes and serotypes of S. mutans isolates. For the serotyping, autoclaved extract antigens from the isolates and serotype-specific antisera against seven known serotypes of S. mutans were prepared. The serotypes of the isolates were demonstrated in immunodiffusion test. In addtition, the prevalence of ${\beta}$-hemolysis on 5% sheep blood agar plate in restricted anaerobic condition and yellow pigment production on 5% sucrose agar plate in less anaerobic condition among the isolates were investigated. The results were as follows: 1. Forty-eight S. mutans strains were isolated from dental plaque samples of 33 patients (62.3%) among 53 patients. 2. Of the isolates, some strains were not grown on MSB medium. 3. Serotype c S. mutans was found in 60.6%, serotype d was found in 30.3% of the patients who were known to harbor S. mutans. 4. Of. the isolates, serotype c isolates were most prevalent (43.8%), serotype d isolates were 25.0%, and serotype b, e, f and g isolates were also found but in lower frequencies. Serotype a S. mutans were not detected. 5. The correlation between serotype and biotype of the isolates was found in 78.6% of the typing cases. 6. Strains revealed ${\beta}$-hemolysis were in 52.1% of the isolates, strains produced yellow pigment were in 47.9% of the isolates, and with one exception, all the strains were belong to serotype c, e and f. 7. The majority of the isolates which revealed ${\beta}$-hemolysis appeared to be yellow pigmented, these isolates were belong to serotype c, e and f.