• 생약학회지
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• 제53권2호
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• pp.87-95
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• 2022

Methicillin resistance Staphylococcus aureus (MRSA) is a gram-positive bacterium, the most commonly isolated bacterial human pathogen. JiYu-san is one of the natural products used to treat diseases in the folk recipe. In this study, we investigated the antimicrobial activity of EtOH 70% extracts of JiYu-san (JYS) against MRSA. The antibacterial activity of JYS against MRSA strain was evaluated using minimum inhibitory concentration (MIC), checkerboard dilution test, and time-kill assay. The effect of JYS on the immune mechanism of MRSA was confirmed through cell membrane permeability tests and energy metabolism tests, and the antibacterial activity mechanism was performed using qRT-PCR and western blot. As a result, in the antibacterial test of JYS, the MIC was measured to be 1.9~1000 ㎍/mL, and synergistic or showed a partial synergistic effect. In addition, JYS showed antibacterial activity in a combination test with DCCD or TX-100. In a study on the mechanism of action of antibacterial activity, it was found that JYS suppressed MRSA resistance genes and proteins. These results suggest that JYS has antibacterial activity and provides great potential as a natural antibiotic by modulating the immune mechanism against MRSA.

• Journal of Microbiology and Biotechnology
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• 제34권6호
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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.

• 대한임상검사과학회지
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• 제38권1호
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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.

• 한국산학기술학회논문지
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• 제16권5호
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• pp.3315-3322
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• 2015

황색포도상구균은 임상에서 중요한 병원체로 원내감염의 주 원인균이다. methicillin에 내성인 황색포도상구균(MRSA) 출현은 특히 전 세계적으로 임상과 역학의 주요 문제가 되고 있으며 다른 병원성 세균과 차별되는 특징으로 독성인자를 가지면서 항생제 내성을 빠르게 획득해 가고 있다. 본 연구는 한 종합병원 임상 검체로부터 얻어진 MRSA 101균주의 독소유전자(enterotoxin(se), toxic shock syndrome toxin-1(tst), exfoliative toxin(et), Panton Valentine leukocidin(pvl)) 보유 분포와 이들 독소 유전자 보유 균주들간의 항생제 내성과의 상관성을 비교 분석 하고자 하였다. 독소유전자중 가장 많이 분리된 seg 유전자가 59균주(58.4%)에서 검출되었으며 70균주(69.3%)에서 두개 이상의 독소 유전자가 검출되었다. 독소유전자 보유조합으로 seb, sec, seg, sei, tst가 19균주(18.8%)에서 발견되었으며 다음으로 빈번한 조합은 sec, seg, sei로 11균주(10.9%)로 나타났다. seb, sec, seg, sei, tst 동일한 독소유전자 조합을 갖고 있는 19균주(18.8%)들은 Ampicillin, Benzylpenicillin, Ciprofloxacin, Clindamycin, Gentamicin, Erythromycin, Telithromycin, Tetracycline 항생제 내성이 100% 였다. 독소유전자 보유수가 많은 균주들은 항생제 내성이 높은 상관성을 확인하였다. 향후 MRSA 균주의 내성이 확산되지 않도록 효과적인 치료와 철저한 감염관리가 선행되어야 할 것이다.

• Journal of Microbiology and Biotechnology
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• 제15권5호
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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.

• 대한수의학회지
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• 제48권1호
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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.

• 대한임상검사과학회지
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• 제48권3호
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• pp.202-209
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• 2016

본 연구는 methicillin-resistant Staphylococcus aureus(MRSA)로부터, 독소 유전자형과 항생제 내성의 상관 관계를 결정하는 것을 목표로 하였다. 2014년 1월~12월까지 전남 순천의 한 병원 중환자실의 임상검체 2,664건에서 얻어진 MRSA 52균주를 분리하였다. 유전자들이 암호화하고 있는 mecA, 장독소(staphylococcal enterotoxins; sea, seb, sec, seg, seh, sei, sej), 독성 쇼크 증상독소-1 (toxic shock syndrome toxin-1; tst-1), 표피박탈성독소(exfoliative toxin; eta, etb), 백혈구 용해 독소(Panton-Valentine leukocidin; pvl)를 특이적 프라이머를 이용한 multiplex PCR로 증폭 검출 하였다. 독소 유전자 seg와 sei 유전자가 각각 40균주(76.9%)로 가장 많은 보유율을 나타냈으며 다음으로 tst 34균주(65.4%) 순으로 검출 되었으며 eta, etb, sea, sed, see, seh, sej와 pvl 유전자들은 검출 되지 않았다. 2개 이상의 독소 유전자를 동시에 보유한 조합의 MRSA는 40균주(76.9%) 였는데 5개 유전자(seb, sec, seg, sei, tst)를 동시 보유한 조합이 28균주(53.8%)로 가장 많은 분포를 보였으며 다음으로 seg, sei 유전자 동시 보유 조합으로 6균주(11.5%)에서 나타났다. 유전자들 간의 동시 보유율은 72.5~100%로서 특정한 독소 유전자 seb, sec, seg, sei와 tst 유전자간의 상관성이 높게 나타났다. 특정 다수의 독소유전자(seb, sec, seg, sei, tst)를 동시에 보유한 균주들이 개별적 독소 유전자를 보유한 균주(seb, sec, tst)와의 항생제 내성의 상관성은 ciprofloxacin, clindamycin, erythromycin 항생제에 100% 내성을 보임으로서 공통적으로 포함된 seb, sec, tst 유전자와 이 항생제의 내성과는 밀접한 연관이 있음을 알았다.

• 대한한방내과학회지
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• 제34권3호
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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.

• 한국축산식품학회지
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• 제40권5호
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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.

• 한국미생물·생명공학회지
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• 제49권3호
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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.