• 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.

• Journal of Microbiology and Biotechnology
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• 제17권4호
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• pp.673-676
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• 2007

A two-step triplex PCR assay targeting the mecA, femA, and nuc genes was developed for the detection of methicillin resistance genes harbored by some Staphylococcus aureus isolates and for the simultaneous identification of such isolates at the species level. The triplex PCR revealed the presence of the femA and nuc genes in all the S. aureus isolates examined (n=105). Forty-four clinical isolates were mecA positive and no foodborne isolates were mecA positive. The PCR results had a 98 or 99% correlation with the results of PBP2a latex agglutination tests or oxacillin susceptibility tests, respectively.

• Bulletin of the Korean Chemical Society
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• 제32권8호
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• pp.2695-2699
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• 2011

${\beta}$ Ketoacyl acyl carrier protein synthase III (KAS III) initiates fatty acid synthesis in bacteria and is a key target enzyme to overcome the antibiotic resistance problem. In our previous study, we found flavonoid inhibitors of Enterococcus faecalis KAS III and proposed three potent antimicrobial flavonoids against Enterococcus faecalis and Vancomycin-resistant Enterococcus faecalis with MIC values in the range of 128-512 ${\mu}g/mL$ as well as high binding affinities on the order from $10^6$ to $10^7\;M^{-1}$. Using these series of flavonoids, we conducted biological assays as well as docking study to find potent flavonoids inhibitors of Staphylococcus aureus KAS III with specificities against Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. Here, we propose that naringenin (5,7,4'-trihydroxyflavanone) and eriodictyol (5,7,3',4'-tetrahydroxyflavanone) are potent antimicrobial inhibitors of Staphylococcus aureus KAS III with binding affinity of $3.35{\times}10^5$ and $2.01{\times}10^5\;M^{-1}$, respectively. Since Arg38 in efKAS III is replaced with Met36 in saKAS III, this key difference caused one hydrogen bond missing in saKAS III compared with efKAS III, resulting in slight discrepancy in their binding interactions as well as decrease in binding affinities. 4'-OH and 7-OH of these flavonoids participated in hydrogen bonding interactions with backbone carbonyl of Phe298 and Ser152, respectively. In particular, these flavonoids display potent antimicrobial activities against various MRSA strains in the range of 64 to 128 ${\mu}M$ with good binding affinities.

• 한방재활의학과학회지
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• 제30권1호
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• pp.31-45
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• 2020

Objectives In this study, we investigated the antimicrobial activity of a 70% ethanol extract of Maneung-hwan (MEH), which is prescribed by practitioners of oriental medicine for use against methicillin-resistant Staphylococcus aureus (MRSA). Methods The antibacterial activity of MEH against MRSA strains was evaluated using the disc diffusion method, broth microdilution method (minimal inhibitory concentration, MIC), checkerboard dilution test, and time-kill test. The mechanism of action of MEH was investigated by bacteriolysis using detergents or ATPase inhibitors Additionally, mRNA and protein expression were investigated by quantitative reverse transcription-polymerase chain reaction and western blot assay, respectively. Results The MIC of MEH was 25~1,600 ㎍/mL against all the tested bacterial strains. We showed that MEH extract exerts strong antibacterial activity. In the checkerboard dilution test, the fractional inhibitory concentration index of MEH in combination with antibiotics indicated synergism or partial synergism against S. aureus. The time-kill study indicated that the growth of the tested bacteria was considerably inhibited after a 24-h treatment with MEH and selected antibiotics. To measure the cell membrane permeability, MEH (3.9 ㎍/mL) was combined with Triton X-100 (TX) at various concentrations N,N-dicyclohexylcarbodimide (DCCD) was also tested as an ATPase inhibitor. TX and DCCD cooperation against S. aureus exhibited synergistic action. Accordingly, the antimicrobial activity of MEH in the context of cell membrane rupture and ATPase inhibition was assessed. Additionally, the expression of genes and proteins associated with resistance was reduced after exposing MRSA to MEH. Conclusions These results suggest that MEH possesses antibacterial activity and acts as a potential natural antibiotic against MRSA.

• 대한의생명과학회지
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• 제25권1호
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• pp.75-82
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• 2019

We developed the multiplex LAMP assay using 16S rRNA, femA and mecA genes for direct detection of the methicillin resistance in Staphylococci from positive blood culture. To simultaneously recognize Staphylococci genus, S. aureus and methicillin resistance, three sets of six primers for 16S rRNA, femA and mecA were designed, respectively. The performance of LAMP assay was affirmed using VITEK system for the phenotypic methods of identification and for oxacillin and cefoxitin antimicrobial susceptibility. The optimal condition for LAMP assay was obtained under $64^{\circ}C$ for 50 min. The detection limit was determined to be of 20 copies and CFU/reaction ($10^4CFU/mL$). For clinical application of comparison with phenotypic methods, the sensitivity and specificity of the LAMP with femA gene for detecting S. aureus was 95.31% and 100%, respectively. The sensitivity and specificity of the LAMP with mecA gene for detecting methicillin resistance was 98.46% and 100%, respectively. The multiplex LAMP assay with femA and mecA gene successfully detected all of MRSA (38 isolates) isolates from 103 Staphylococci in blood cultures. The LAMP assay developed in this study is sensitive, specific, and of excellent agreement with the phenotypic methods.

• 한국식품위생안전성학회지
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• 제36권6호
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• pp.520-527
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• 2021

본 연구에서는 보육시설 실내공기에서 분리된 식중독 균주의 독소 유전자 분포와 항생제 내성을 분석하여 보육시설 실내공기에 의한 식중독 발생을 사전 예방하고 식중독 발생 시 적절한 치료를 위한 기초자료를 제공하고자 하였다. 어린이집 실내공기에서 분리된 Staphylococcus aureus 16주, Bacillus cereus 37주를 실험대상으로 하였다. S. aureus와 B. cereus 독소 유전자는 PCR 방법으로 검출하였다. 항생제 감수성 실험은 Clinical and Laboratory Standard Institute의 디스크 확산법에 따라 실험하였다. S. aureus 16 균주 중 11 균주(68.6%)에서 seg와 sei 독소 유전자가 검출되었다. B. cereus 37 균주 모두에서 nheA와 nheB 독소 유전자가 검출되었다. B. cereus 독소 유전자 패턴은 총 12개로 나타났으며 nheA-nheB-nheC 독소 유전자가 가장 중요한 패턴으로 나타났다. S. aureus 16 균주의 항생제 감수성실험 결과 ampicillin과 penicillin 항생제에 93.8%, 87.5% 내성을 나타내었으나 methicillin resistance Staphylococcus aureus와 vacomycin resistance Staphylococcus aureus는 검출되지 않았다. B. cereus 37 균주의 항생제 감수성 실험 결과 ampicillin과 penicillin 항생제에 100% 내성을 나타냈었다. 이러한 결과를 종합하여 볼 때 보육시설 실내공기에 오염된 S. aureus와 B. cereus에 의한 식중독을 발생을 예방하기 위하여 주기적인 환기와 공기 질 관리가 필요한 것으로 판단되었다.

• 대한본초학회지
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• 제28권1호
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• pp.59-64
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• 2013

Objectives : Methicillin-Resistant Staphylococcus aureus (MRSA) is a cephalosporin and beta-lactam antibiotic-resistant strains. In most cases, that is spread from infected patients and infection rates are growing increasingly. Thus, accordingly, 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 infections diseases. Methods : The antibacterial activities of Sinhyowoldosan were evaluated against 3 strains of Methicillin-resistant staphylococcus aureus(MRSA) and 1 standard Methicillin-susceptible S. aureus (MSSA) strain by using the disc diffusion method, minimal inhibitory concentrations (MICs) assay, colorimetric assay using MTT test, checkerboard dilution test and time-kill assay was performed under dark. Results : The MIC (minimum inhibitory concentration) of Sinhyowoldosan water extract against S. aureus strains ranged from 500 to 2,000 ${\mu}g/mL$, so we have confirmed it on a strong antibacterial effect. Also, the combinations of Sinhyowoldosan water extract and conventional antibiotics exhibited improved inhibition of MRSA with synergy effect. We suggest that Sinhyowoldosan water extract against MRSA have antibacterial activity, it has potential as alternatives to antibiotic agent. the combination test was used, Triton X-100 (TX) and DCCD for measurement of membrane permeability and inhibitor of ATPase. As a result, antimicrobial activity of SH is affected by the cell membrane were assessed. Conclusion : We suggest that the Sinhyowoldosan 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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• 제43권5호
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• pp.792-804
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• 2023

Non-aureus staphylococci (NAS), particularly antimicrobial-resistant NAS, have a substantial impact on human and animal health. In the current study, we investigated (1) the species profiles of NAS isolates collected from healthy broilers, farm environments, and farm workers in Korea, (2) the occurrence of antimicrobial-resistant NAS isolates, especially methicillin resistance, and (3) the genetic factors involved in the methicillin and fluoroquinolone resistance. In total, 216 NAS isolates of 16 different species were collected from healthy broilers (n=178), broiler farm environments (n=18), and farm workers (n=20) of 20 different broiler farms. The two most dominant broiler-associated NAS species were Staphylococcus agnetis (23.6%) and Staphylococcus xylosus (22.9%). Six NAS isolates were mecA-positive carrying staphylococcal cassette chromosome mec (SCCmec) II (n=1), SCCmec IV (n=1), SCCmec V (n=2), or nontypeable SCCmec element (n=2). While two mecA-positive Staphylococcus epidermidis isolates from farm workers had SCCmec II and IV, a mecA-positive S. epidermidis isolate from broiler and a Staphylococcus haemolyticus isolate farm environment carried SCCmec V. The occurrence of multidrug resistance was observed in 48.1% (104/216 isolates) of NAS isolates with high resistance rates to β-lactams (>40%) and fusidic acid (59.7%). Fluoroquinolone resistance was confirmed in 59 NAS isolates (27.3%), and diverse mutations in the quinolone resistance determining regions of gyrA, gyrB, parC, and parE were identified. These findings suggest that NAS in broiler farms may have a potential role in the acquisition, amplification, and transmission of antimicrobial resistance.

• 대한수의학회지
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• 제39권2호
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• pp.376-382
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• 1999

A methicillin-resistant Staphylococcus aureus (MRSA) isolate was recovered from a 9-month-old female Shih-Tzu dog with canine distemper virus infection. We performed in vitro antimicrobial susceptibility test to determine the most effective antimicrobial drug against the isolate and thus, to emphasize its potential clinical importance in animal practices. Isolate was confirmed MRSA by oxacillin agar screening test. The isolate was fully resistant to all $\beta$-lactam antibiotics and was susceptible to glycopeptides. Of the other antibiotics, mupirocin, TMP/SMZ (trimethoprim-sulfamethoxazole), and chloramphenicol showed inhibitory effect at the concentration of 4x MIC. The MICs ranged 0.25->$128{\mu}g/ml$, and MBCs ranged 0.5->$128{\mu}g/ml$. The combined TMP/SMZ with cefamandole or novobiocin showed synergistic effect, whereas the combination of novobiocin plus cefamandole or teicoplanin resulted in antagonistic effects. Although MRSA in animals so far has been reported in the geographically limited countries, at least theoretically, it could be occurred in the future more frequently through either human or animal origin. The use of this combination may be of value in this situation. As with all antimicrobial agents, inappropriate or unnecessarily prolonged therapy may contribute to the emergence of resistance strains and loss of efficacy.

• Journal of Microbiology and Biotechnology
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• 제31권9호
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• pp.1288-1294
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• 2021

There are a growing number of reports of hospital-acquired infections caused by pathogenic bacteria, especially methicillin-resistant Staphylococcus aureus (MRSA). Many plant products are now being used as a natural means of exploring antimicrobial agents against different types of human pathogenic bacteria. In this research, we sought to isolate and identify an active molecule from Sedum takesimense that has possible antibacterial activity against various clinical isolates of MRSA. NMR analysis revealed that the structure of the HPLC-purified compound was 1,2,4,6-tetra-O-galloyl-glucose. The minimum inhibitory concentration (MIC) of different extract fractions against numerous pathogenic bacteria was determined, and the actively purified compound has potent antibacterial activity against multidrug-resistant pathogenic bacteria, i.e., MRSA and its clinical isolates. In addition, the combination of the active compound and β-lactam antibiotics (e.g., oxacillin) demonstrated synergistic action against MRSA, with a fractional inhibitory concentration (FIC) index of 0.281. The current research revealed an alternative approach to combating pathogenesis caused by multi-drug resistant bacteria using plant materials. Furthermore, using a combination approach in which the active plant-derived compound is combined with antibiotics has proved to be a successful way of destroying pathogens synergistically.