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

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

• 한국식품과학회지
• /
• 제37권3호
• /
• pp.456-464
• /
• 2005

본 연구에서는 최근 전세계적으로 문제가 되고 있는 항생제 내성균에 대해 그 항균효과를 천연의 약용식물을 이용하여 분자생물학적 수준에서 비교해보고 천연항균제의 개발에 우선하여 직접 식품에 첨가하여 그 효과를 검색하고자 하였다. 병원내감염의 주된 원인균으로 알려진 황색포도상구균(Staphylococcusaureus)은 여러 항균제에 대하여 내성을 획득함으로써 질병의 치료를 어렵게 한다는 점에서 매우 심각한 문제이다. 이러한 내성균은 다양한 방법으로 동 식물에 빠르게 전파되고 있으므로 소비자가 내성균을 지닌 식품을 섭취해 잠재적인 내성균을 보유하게 되는 결과를 낳게 된다. 이러한 항생제 내성 황색포도상구균(Methicillin Resistant S. aureus, MRSA)에 대해 천연 약용 식물 중 우수한 항균력을 가지는 감초를 순차 분획하여 각 분획물을 이용한 MRSA에 대한 항균 활성을 알아본 결과 hexane, chloroform 분획물에서 항균활성을 나타냈으며, 동일농도의 생육저해비교 실험에서 대부분의 내성균주에 대해 대조군과 유사한 효과를 나타낸 penicillin에 비해 감초분획물이 대수기를 넘는 억제효과를 나타내었으며 이 중 chloroform 분획물이 가장 높은 저해활성을 나타내었다. 또한 RT-PCR을 통하여 표준균주와 내성균주의 내성 정도와 내성 유전자를 관찰해본 결과 KCCM 40510은 내성획득유전자인 mecA, mecRI, mecI, femA에 발현이 높게 나타나 고도내성균임을 알 수 있었으며, 고도내성을 나타내는 균주를 이용하여 감초의 chloroform 분획물을 농도별로($50,\;100,\;250\;{\mu}/mL$) 처리한 결과 농도 의존적으로 유전자 발현이 억제됨을 볼 수 있었다. 특히 고도내성균주인 KCCM 40510은 메티실린 내성을 상승시키는 mecA 유전자의 발현을 억제시키는 mecRI 유전자의 pathway와 상응하는 것으로 보여지므로 감초가 이러한 고도내성균에 효과가 있음을 알 수 있었다. 또한 SEM을 통한 형태학적 관찰을 통해 분획물 처리 시 세포벽이 파괴되는 것을 확인할 수 있었다. 한편 이러한 감초추출물을 직접 식품에 첨가하여 내성 균주에 대한 억제 효과도 확인한 결과 추출물 200mg/100mL 농도에서 포도상구균에 대해 항균효과를 나타내는 것을 볼 수 있었다.

• Tuberculosis and Respiratory Diseases
• /
• 제65권2호
• /
• pp.91-98
• /
• 2008

연구배경: 메티실린 내성 황색포도알균은 의료기관 관련 감염 뿐 아니라 지역사회 감염에서도 나타나 이에 저자들은 한 대학병원 중환자실에 입원한 환자의 비강 도말 배양을 통해 MRSA의 의료기관 관련 감염 및 지역사회 감염의 빈도와 분자 역학 및 항생제 내성을 연구하였다. 방 법: 2006년 6월에서 9월까지 건양대학교 병원 중환자실 환자 353명을 대상으로 입실 첫날 비강 도말 배양을 시행하여 MRSA 획득 위험인자에 따라 HA-MRSA와 CA-MRSA로 나누어 Pulsed-Field Gel Electrophorosis (PFGE)로 분류하여 각각의 항생제 내성 검사를 시행하였다. 결 과: 353명 중 비강 도말 배양에서 동정된 MRSA는 42명(11.9%)이며, 동정된 MRSA 중 HA-MRSA는 33명(78.6%), CA-MRSA는 9명(21.4%)이다. PFGE에서 type A에서 type K까지 11형으로 구분하였고 HA-MRSA는 type A (n=9), B (n=7)가, CA-MRSA는 type A (n=2), B (n=2)가 주로 나타났다. 항생제 내성률은 erythromycin, ciprofloxacin에서 HA-MRSA가 CA-MRSA보다 높게 나타났다. 결 론: MRSA의 집락률은 11.9%이며 HA-MRSA의 균주가 CA-MRSA보다 많고, CA-MRSA는 9예로 적은 예지만 PFGE type에서 대부분의 type이 HA-MRSA에서 동정된 type과 같은 경향을 보여 지역사회전파를 시사한다.

• Journal of Preventive Medicine and Public Health
• /
• 제45권3호
• /
• pp.174-180
• /
• 2012

Objectives: An ambulance can be a potential source of contagious or droplet infection of a community. We estimated the prevalence of positive carriage of tuberculosis (TB), methicillin-resistant $Staphylococcus$ $aureus$ (MRSA), and vancomycin-resistant $Enterococci$ (VRE) in patients transported by ambulance. Methods: This was a retrospective observational study. We enrolled all patients who visited a tertiary teaching hospital emergency department (ED). Blood, sputum, urine, body fluid, and rectal swab samples were taken from patients when they were suspected of TB, MRSA, or VRE in the ED. The patients were categorized into three groups: pre-hospital ambulance (PA) group; inter-facility ambulance (IA) group; and non-ambulance (NA) group. Adjusted odds ratio (OR) and 95% confidence intervals (CI) were calculated using a multivariable logistic regression model for the prevalence of each infection. Results: The total number of patients was 89206. Of these, 9378 (10.5%) and 4799 (5.4%) were in the PA and IA group, respectively. The prevalence of TB, MRSA, and VRE infection were 0.3%, 1.1%, and 0.3%, respectively. In the PA group, the prevalence of TB, MRSA, and VRE were 0.3%, 1.8%, and 0.4%. In the IA group, the prevalence of TB, MRSA, and VRE were 0.7%, 4.6%, and 1.5%, respectively. The adjusted ORs (95% CI) of the PA and IA compared to the NA group were 1.02 (0.69 to 1.53) and 1.83 (1.24 to 2.71) for TB, 2.24 (1.87 to 2.69) and 5.47 (4.63 to 6.46) for MRSA, 2.59 (1.78 to 3.77) and 8.90 (6.52 to 12.14) for VRE, respectively. Conclusions: A high prevalence of positive carriage of TB, MRSA, and VRE in patients transported by metropolitan ambulances was found.

• 한방재활의학과학회지
• /
• 제25권2호
• /
• pp.1-13
• /
• 2015

Objectives Methicillin-Resistant Staphylococcus aureus (MRSA) is a human pathogen and a major cause of hospital-acquired infections. New antibacterial agents that have not been compromised by bacterial resistance are needed to treat MRSA-related infections. In this study, we investigated the antimicrobial activity ofethanol extract of Haedokgeumhwa-san (HGH) which prescription is composed of korean medicine against MRSA. Methods The antibacterial activity of HGH extract was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method (minimal inhibitory concentration; MIC), checkerboard dilution test, and time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors. The checkerboard dilution test was used to examined synergistic effect of ampicillin, oxacillin, ciprofloxacin, vancomycin, gentamicin and norfloxacin in combination with HGH ethanol extract. A time-kill assay was performed a survival curve which was obtained by plotting viable colony counts depending on time on bacterial growth. Results The minimum inhibitory concentration (MIC) of ethanol extract (HGH) ranged from 1,000 to $2,000{\mu}g/mL$ against all the tested bacterial strains, respectively. We are able to confirm that HGH extract has potentially strong antibacterial activity. In the checkerboard dilution test, fractional inhibitory concentration index of HGH in combination with antibiotics indicated synergy or partial synergism against S. aureus. A time-kill study showed that the growth of the tested bacteria was considerably inhibited after 8 hr of treatment with the combination of HGH with selected antibiotics. For measurement of cell membrane permeability, HGH $250{\sim}1,000{\mu}g/mL$ along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (Tris) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide ($NaN_3$) was used as an inhibitor of ATPase. TX, Tris, DCCD and $NaN_3$ cooperation against S. aureus showed synergistic action. Accordingly, antimicrobial activity of HGH was affected by cell membrane and inhibitor of ATPase. Conclusions These results suggest that Haedokgeumhwa-san extract has antibacterial activity, and that HGH extract offers a potential as a natural antibiotic against MRSA.

• 생명과학회지
• /
• 제21권4호
• /
• pp.589-595
• /
• 2011

질병를 유발하는 미생물들의 항생물질에 대한 증가하는 내성문제를 극복할 수 있는 새로운 항생물질을 개발하고자 항생물질을 생산하는 방선균을 탐색하여 특성화하는데 본 연구는 초점을 맞추고 있다. 충청북도 일대의 토양으로부터 PDA, starch casein agar 등의 배지를 이용하여 100여 개의 방선균주를 순수분리하였다. 순수분리된 방선균주 중 메티실린 내성 황색포도상구균에 가장 활성이 뛰어난 BCNU 1030을 선발하여 형태학적, 생화학적, 분자생물학적 방법으로 동정한 결과 Streptomyces 속으로 동정되었다. 분리균주인 BCNU 1030은 MRSA를 포함한 그람양성 및 그람음성세균에 대한 폭넓은 항균스펙트럼을 가졌다. BCNU 1030의 디클로로메탄 추출물은 MRSA CCARM 3090에 대하여 최소$0.78\;{\mu}g/ml$의 농도에서 저해활성이 있음이 확인되었다. 그러므로 BCNU 1030은 새로운 anti-MRSA 항생물질의 개발에 있어서 가능성을 가지고 있는 것으로 판단된다.

• Journal of Veterinary Science
• /
• 제21권4호
• /
• pp.54.1-54.11
• /
• 2020

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a leading cause of severe infections in humans and animals worldwide. Studies elucidating the population structure, staphylococcal cassette chromosome mec types, resistance phenotypes, and virulence gene profiles of animal-associated MRSA are needed to understand spread and transmission. Objectives: The objective of this study was to determine 1) clonal complexes and spa types, 2) resistance phenotypes, and 3) virulence/resistance gene profiles of MRSA isolated from animals in Switzerland. Methods: We analyzed 31 presumptive MRSA isolates collected from clinical infections in horses, dogs, cattle, sheep, and pigs, which had tested positive in the Staphaurex Latex Agglutination Test. The isolates were characterized by spa typing and DNA microarray profiling. In addition, we performed antimicrobial susceptibility testing using the VITEK 2 Compact system. Results: Characterization of the 31 presumptive MRSA isolates revealed 3 methicillinresistant Staphylococcus pseudintermedius isolates, which were able to grow on MRSA2 Brilliance agar. Of the 28 MRSA isolates, the majority was assigned to CC398 (86%), but CC8 (11%) and CC1 (4%) were also detected. The predominant spa type was t011 (n = 23), followed by t009 (n = 2), t034 (n = 1), t008 (n = 1), and t127 (n = 1). Conclusions: The results of this study extend the current body of knowledge on the population structure, resistance phenotypes, and virulence and resistance gene profiles of MRSA from livestock and companion animals.

• Journal of Microbiology and Biotechnology
• /
• 제20권3호
• /
• pp.637-642
• /
• 2010

We investigated the prevalence and the molecular characteristics of vancomycin-intermediate Staphylococcus aureus (VISA) among methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from clinical samples at tertiary or general hospitals participating in a nationwide surveillance program for VISA and vancomycin-resistant Staphylococcus aureus (VRSA) in Korea during an 8-week period in each year from 2001 to 2006. Of 41,639 MRSAs isolated, 37,856 were screened and 169 grew on brain heart infusion agar supplemented with 4 ${\mu}g/ml$ vancomycin. A vancomycin MIC of 4 ${\mu}g/ml$ was confirmed for 33 VISA isolates of the 169 isolates. Eighteen of the 33 isolates were classified as hetero-VISA (hVISA) by the population analysis profile (PAP) method. All VISA isolates were susceptible to linezolid, tigecycline, and quinupristin-dalfopristin. Most VISA isolates (MIC 4 ${\mu}g/ml$) showed a PFGE C pattern with sec, seg, and sei enterotoxin genes, including ST5-SCCmec type II, or a PFGE A pattern with sea, including ST239-SCCmec type III.

• Journal of Microbiology and Biotechnology
• /
• 제31권8호
• /
• pp.1060-1068
• /
• 2021

Community-associated Methicillin-Resistant Staphylococcus aureus (CA-MRSA) is notorious as a leading cause of soft tissue infections. Despite several studies on the Agr regulator, the mechanisms of action of Agr on the virulence factors in different strains are still unknown. To reveal the role of Agr in different CA-MRSA, we investigated the LACΔagr mutant and the MW2Δagr mutant by comparing LAC (USA300), MW2 (USA400), and Δagr mutants. The changes of Δagr mutants in sensitivity to oxacillin and several virulence factors such as biofilm formation, pigmentation, motility, and membrane properties were monitored. LACΔagr and MW2Δagr mutants showed different oxacillin sensitivity and biofilm formation compared to the LAC and MW2 strains. Regardless of the strain, the motility was reduced in Δagr mutants. And there was an increase in the long chain fatty acid in phospholipid fatty acid composition of Δagr mutants. Other properties such as biofilm formation, pigmentation, motility, and membrane properties were different in both Δagr mutants. The Agr regulator may have a common role like the control of motility and straindependent roles such as antibiotic resistance, biofilm formation, change of membrane, and pigment production. It does not seem easy to control all MRSA by targeting the Agr regulator only as it showed strain-dependent behaviors.