• Journal of Microbiology and Biotechnology
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• v.34 no.6
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• pp.1322-1327
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• 2024

The accurate and rapid detection of methicillin-resistant Staphylococcus aureus (MRSA) holds significant clinical importance. This work presents a new method for detecting methicillin-resistant Staphylococcus aureus (S. aureus) in clinical samples. The method uses an aptamer-based colorimetric assay that combines a recognizing probe to identify the target and split DNAzyme to amplify the signal, resulting in a highly sensitive and direct analysis of methicillin-resistance. The identification of the PBP2a protein on the membrane of S. aureus in clinical samples leads to the allosterism of the recognizing probe, and thus provides a template for the proximity ligation of split DNAzyme. The proximity ligation of split DNAzyme forms an intact DNAzyme to identify the loop section in the L probe and generates a nicking site to release the loop sequence ("3" and "4" fragments). The "3" and "4" fragments forms an intact sequence to induce the catalytic hairpin assembly, exposing the G-rich section. The released the G-rich sequence of LR probe induces the formation of G-quadruplex-hemin DNAzyme as a colorimetric signal readout. The absorption intensity demonstrated a strong linear association with the logarithm of the S. aureus concentration across a wide range of 5 orders of magnitude dynamic range under the optimized experimental parameters. The limit of detection was calculated to be 23 CFU/ml and the method showed high selectivity for MRSA.

• Korean Journal of Clinical Laboratory Science
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• v.38 no.1
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• pp.38-44
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• 2006

A simple and easy modification of AST by disk diffusion was tested for the detection of induced clindamycin resistant Staphylococci and their antimicrobial susceptibility at the same time. The incidence of inducible clindamycin resistant staphylococci in blood culture and their MIC characterization at Seoul National University Hospital was analyzed by an AST contained disk approximation test (D-zone test) and Etest, respectively. Of the total 309 staphylococcal isolates, 139 (45%) isolates presented constitutive resistance to ERY and CLI (ERY-R, CLI-R phenotype), and 59 were ERY-I/R and CLI-S phenotypes. Of the 59 isolates, 19 (32%) isolates were inducible resistant to CLI. The incidence was higher in S. aureus (66.7%) than coagulase-negative staphylococci (CNS, 26.0%). Especially, methicillin-resistant staphylococci (MRSA, 100%; MRCNS, 45.5%) presented higher inducibility than methicillin susceptible (MSSA, 50%; MSCNS, 20%). For most of the inducible clindamycin resistant staphylococci (15 of 19 isolates), their ERY MIC were high (>$128_{\mu}g/mL$) and were methicillin resistant. The remaining 4 isolates were methicillin susceptible and their ERY MIC were of intermediate concentrations ($1-4_{\mu}g/mL$). We concluded that suscetibility testing of staphylococci, especially methicillin resistant, should include the D-zone test.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.16 no.5
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• pp.3315-3322
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• 2015

Staphylococcus aureus is the major causative organism of nasocomial infection being the important pathogen in the clinic. Appearance of staphylococcus aureus resistant to methicillin (MRSA) is becoming a big problem in clinics and dynamics all over the world acquiring antibiotic resistance with virulence factors as its feature differentiated from other pathogenic bacteria fast. This research intended to compare and analyze the correlation of antibiotics resistance between strains with toxin genes and distribution of toxin genes of MRSA 101 strains acquired from clinical specimen in one general hospital (enterotoxin(se), toxic shock syndrome toxin-1(tst), exfoliative toxin(et), Panton Valentine leukocidin(pvl)). seg gene, isolated the most among toxin genes, was detected in 59 strains (58.4%) and more than two toxin genes were detected in 70 strains (69.3%). As a combination possessing toxin genes, it was detected in 19 strains (18.8%) as seb, sec, seg, sei, tst and the second frequent combination was sec, seg, sei shown in 11 strains (10.9%). 19 strains (18.8%) with combinations of toxin genes same with seb, sec, seg, sei, tst had 100% resistance Ampicillin, Benzylpenicillin, Ciprofloxacin, Clindamycin, Gentamicin, Erythromycin, Telithromycin, Tetracycline antibiotics. Strains with many toxin genes showed high correlation of antibiotic resistance. Afterwards, effective therapy and thorough infection management should be preceded not to spread the resistance of MRSA strain.

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.1039-1046
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• 2005

The salt resistance of antibacterial activity and synergistic effect with clinically used antibiotic agents are critical factors in developing effective peptide antibiotic drugs. For this reason, we investigated the resistance of antibacterial activity to antagonism induced by NaCl and $MgCl_2$ and the synergistic effect of P20 with cefotaxime. P20 is a 20-residue synthetic peptide derived from a cecropin A (CA)-melittin(ME) hybrid peptide. In this study, P20 was found to have potent antibacterial activity against clinically isolated methicillin-resistant Staphylococcus aureus (MRSA) strains without hemolytic activity against human erythrocytes. The combination study revealed that P20 in combination with cefotaxime showed synergistic antibacterial activity in an energy-dependent manner. We also confirmed the synergism between P20 and cefotaxime by fluorescence-activated flow cytometric analysis by staining bacterial cells with propidium iodide (PI) and bis-(1,3-dibutylbarbituric acid) trimethine oxonol (BOX). This study suggests that P20 may be useful as a therapeutic antibiotic peptide with synergistic effect in combination with conventional antibiotic agents.

• Korean Journal of Veterinary Research
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• v.48 no.1
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• pp.75-81
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• 2008

The rapid evolution of antibiotic resistance in Staphylococcus (S.) aureus is of considerable concern. Methicillin-resistant S. aureus (MRSA) strains are especially one of the greatest public concerns since the treatment of infections is more difficult when encountering resistance. In this study, we conducted a nationwide survey on the antimicrobial resistance of S. aureus isolated from raw meat samples collected from 16 countries, including Korea, and investigated the prevalence of MRSA as a possible source of human infection. Of 1,984 meat samples, S. aureus was isolated from 218 (11.0%) samples consisting of 23 (12.1%) from domestic meat and 195 (10.9%) from imported meat. The isolation rates of poultry meat, pork and beef were 12.8%, 7.0% and 10.0%, respectively. With regard to imported meat, the incidence varied from 4.8% to 16.6% from 13 countries, with the exception of Austria and Poland. In a resistance test to 20 antimicrobial agents, one hundred and eighty-four isolates (84.4%) were resistant to one or more antimicrobial agents tested. Especially, 17 (7.8%), 124 (56.9%) and 28 (12.8%) isolates showed a resistance to 3, 2 and 1 drugs, respectively. One isolate originating from domestic beef was resistant to 7 drugs. Another isolate originating from imported poultry meat showed resistance to oxacillin and methicillin by the disk diffusion test and minimal inhibition concentration methods, but showed negative for detection of the mecA gene.

• Korean Journal of Clinical Laboratory Science
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• v.48 no.3
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• pp.202-209
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• 2016

This study is aimed to determine the correlation between the toxin gene types and antibiotic resistance from MRSA (methicillin-resistant Staphylococcus aureus). Fifty-two strains of MRSA, between January 2014, and December 2014, were isolated from clinical specimens obtained from 2,664 cases in the intensive care unit of a hospital in Suncheon, Jeonnam, Korea. Genes encoding mecA, enterotoxin (SE), toxic shock syndrome toxin-1 (TSST-1), exfoliative toxin (ET), and Panton-Valentine leukocidin (PVL) were detected by multiplex PCR-mediated amplification using specific primers. Toxin genes (seg and sei) were present in 40 strains (76.9%), followed by tst in 34 strains (65.4%). Other genes (eta, etb, sea, sed, see, seh, sej, and pvl) were not detected. Forty strains (76.9%) of MRSA had 2 or more toxin genes simultaneously; 5 coexistent toxin-genes (seb, sec, seg, sei, tst) were the most common in 28 strains (53.8%), and 6 strains (11.5%) had seg and sei genes. The coexistence of genes were 72.5~100%, showing a high correlation among genes (seb, sec, seg, sei and tst). As strains (seb, sec, tst) that had particular toxin genes (seb, sec, seg, sei, tst) in multiple showed 100% resistance to ciprofloxacin, clindamycin, erythromycin, we were able to find that seb, sec, and tst genes have a close relationship to the aforementioned antibiotics. It showed a higher resistance to ciprofloxacin, clindamycin, erythromycin, and tetracycline compared with strains that had toxin genes independent from multiple toxin genes.

• The Journal of Internal Korean Medicine
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• v.34 no.3
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• pp.278-288
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• 2013

Objectives : Methicillin-resistant Staphylococcus aureus (MRSA) has a cephalosporin and beta-lactam antibiotic-resistant strains. MRSA is one of the major pathogens causing hospital infection and the isolation ratio of MRSA has gradually increased. Consequently, increased resistance to antibiotics is causing serious problems in the world. Therefore, there is a need to develop alternative antimicrobial drugs for the treatment of infectious diseases. Methods : The antibacterial activities of Ipyo-san were evaluated against 2 strains of MRSA and 1 standard Methicillin-susceptible staphylococcus aureus (MSSA) strain by using the disc diffusion method, minimal inhibitory concentrations (MIC) assay, colorimetric assay using MTT test, checkerboard dilution test and time-kill assay performed under dark. Results : The MIC of Ipyo-san water extract against S. aureus strains ranged from 1000 to $2,000{\mu}g/ml$, so we confirmed that it had a strong antibacterial effect. Also, the combinations of Ipyo-san water extract and conventional antibiotics exhibited improved inhibition of MRSA with synergy effect. We suggest that Ipyo-san water extract against MRSA has antibacterial activity so it has potential as alternatives to antibiotic agents. For the combination test, we used Triton X-100 (TX) and DCCD for measurement of membrane permeability and inhibitor of ATPase. As a result, antimicrobial activity of Ipyo-san water extract was affected by the cell membrane. Conclusions : We suggest that the Ipyo-san water extract lead the treatment of bacterial infection to solve the resistance and remaining side-effect problems that are the major weak points of traditional antibiotics.

• 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.

• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.449-457
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• 2021

The aim of the present study was to survey the frequency of inducible and constitutive phenotypes and inducible cross-resistant genes by regulating the methylation of 23S rRNA (ermA, ermB, and ermC) and macrolide efflux-related msrA gene in Staphylococcus aureus and S. epidermidis strains. A total of 172 bacterial isolates (identified based on standard tests), were examined in this study. Antibiotic susceptibility was determined by the disk diffusion method, and all isolates were evaluated with respect to inducible and constitutive phenotypes. The presence of ermA, ermB, ermC, and msrA genes was investigated by a PCR assay. The constitutive resistance phenotypes showed a higher distribution among the isolates. R phenotype was detected more among S. epidermidis isolates (46.25%). ermB, ermC, and msrA genes were detected more in methicillin-resistant S. aureus (MRSA) and methicillin-resistant S. epidermidis (MRSE) isolates that had R and HD phenotypes (>77% strains). The ermA gene had the lowest frequency among MRSA, MRSE, MSSA, and MSSE strains (<14% isolates). Distribution of inducible resistance genes in MRSA and MRSE strains, and possibly other species, leads to increased constitutive resistance to erythromycin, clindamycin, and other similar antibiotics. Therefore, it can be challenging to treat infections caused by these resistant strains.

• Korean Journal of Plant Resources
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• v.29 no.6
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• pp.699-703
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• 2016

Methicillin-resistant Staphylococcus aureus (MRSA) is a serious clinical and an urgent problem worldwide. Few new drugs are available against MRSA, because MRSA has the ability to acquire resistance to most antibiotics, which consequently increases the cost of medication. In the present study, the antibacterial activity of Hydnocarpi Semen was investigated. The most effective method is to develop antibiotics from the natural products without having any toxic or side effects. Therefore, there is a need to develop alternative antibacterial drugs for the treatment of infectious diseases. Five Clinical isolates (MRSA) were obtained from five different patients at Wonkwang University Hospital (Iksan, South Korea). The Other 2 strains were ATCC 33591 (Methicillin-resistant strain) and ATCC 25923 (Methicillin-susceptible strain). Antibacterial activity (Minimal Inhibitory Concentrations, MICs) was determined by broth dilution method, disk diffusion method, MTT test, and checkerboard dilution test. Antibacterial activity of n-hexane fraction was remarkable, and had a MICs ranging from $31.25-125{\mu}g/m{\ell}$. FICI values for HFH+AM and HFH+OX were 0.13-0.19 and 0.04-0.29, showing the increase of synergistic effect. When combined together, these antibacterial effects were dramatically increased.