• 제목/요약/키워드: mec A gene

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Identification of Methicillin-Resistant Staphylococcus aureus by Polymerase Chain Reaction (중합효소 연쇄반응을 이용한 메치실린 내성균주의 동정)

  • Park, In-Cheol;Kim, Gwang-Su;Park, Myeong-Jin;Lee, Seung-Hun;Hong, Seok-Il;Choe, Tae-Bu
    • KSBB Journal
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    • v.14 no.4
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    • pp.460-464
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    • 1999
  • Methicillin-resistant Staphyloccus aureus (MRSA) has been known to be resistant to many kinds of antibiotics and causes a problem of nosnocomial infection since the third generation of cephalosporines has been introduced in the 1980s. As antibiotic sensitivity tests which have been routinely used to detect MRSA in the laboratory depend on the culture conditions such as, pH, temperature, and time, etc., it is difficult to decide in the case of borderline- or low-level of MRSA. Therefore it would be necessary to develope a new method based on the molecular biological technique to overcome these problems. In this study, we extracted DNA from S. aureus and performed polymerase chain reaction (PCR) to amplify mec A gene, encoding penicillin-binding protein 2' (PBP-2'), which is known to confer bacteria resistance to the bacteriostatic action of methicillin. The results were compares with those of minimal inhibitory concentration (MIC) test. When MIC test with oxacillin was performed on the 120 isolates of S. aureus from each patient's specimens, 64 of them were MRSA and 56 of them were methicillin-sensitive Staphylococcus aureus (MSSA). In pus specimen, more precisely, 61.9% (26/42) of MRSA was detected, and 44.2% (19/43), 60% (9/15) and 50% (10/20) of MRSA were detected in sputum, body fluid, and other specimen respectively. When 40 isolates of MRSA and MSSA were tested by PCR method and compares with the results of MIC method, different results were obtained from 1 isolate of MRSA (2.5%) and in 2 isolates of MSSA (5%) suggesting that PCR method should be performed at the same time for more accurate clinical test of MRSA.

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The Correlation between Toxin Genotype and Antibiotic Resistance in Methicillin Resistant Staphylococcus aureus Isolated from Clinical Specimen of Intensive Care Unit (중환자실의 임상검체로부터 분리된 Methicillin 내성 Staphylococcus aureus의 독소유전자형과 항생제내성의 상관관계)

  • Park, Chul;Seong, Chi Nam
    • 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.

Prevalence of Methicillin-resistant Staphylococci Isolates from Horses and Horse-related Personnel in Korea (말 및 말관련 종사자의 methicillin 내성 포도상구균의 유병율 조사)

  • Lee, Sang-Kyu;Han, Jae-Ik;Kim, Ill-Hwa;Na, Ki-Jeong;Kang, Hyun-Gu
    • Journal of Veterinary Clinics
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    • v.31 no.3
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    • pp.194-198
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    • 2014
  • Methicillin-resistant staphylococci (MRS) are emerging as important pathogens in humans and animals worldwide. The aim of this study was to investigate the prevalence of MRS in the racehorse population and in horse-related personnel in Korea. A total of 195 horses and 18 humans (eight veterinarians, three veterinary hospital staff, and seven horse-handlers) from racehorse farms in Korea were included in the study. The samples were collected from nasal cavities using bacterial transport medium and were cultivated on tryptic soy agar with 5% sheep blood for 3 days at $37^{\circ}C$ to confirm the presence of Staphylococcus spp. Presumptive Staphylococcus spp. isolates were identified by 16S ribosomal RNA gene analysis. The coagulase test and oxacillin susceptibility tests were performed using the tube dilution and disk diffusion methods, respectively. The presence of the mecA gene was determined using a polymerase chain reaction assay. Of the 195 horses, 29 (15.6%) yielded 29 MRS isolates. Twelve (66.7%) of the 18 horse-related personnel yielded 12 MRS isolates. All of the MRS isolates from horses or horse-related personnel were identified as methicillin-resistant coagulase-negative staphylococci (MRCNS). The result of this study suggest that the prevalence of MRS increased with the duration of antibiotic use (p = 0.002). This study also provides evidence for the zoonotic transmission of MRCNS between horses and humans, although further investigations are needed.

Virulence Factors of Staphylococcus aureus Isolated from Korean Pork bulgogi: Enterotoxin Production and Antimicrobial Resistance

  • Jung, Byeong Su;Lee, Yong Ju;Lee, Na-Kyoung;Kim, Hyoun Wook;Oh, Mi-Hwa;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.35 no.4
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    • pp.502-506
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    • 2015
  • The aim of this study was to investigate the antimicrobial resistance profiles of and the enterotoxin gene distribution in 4 strains of Staphylococcus aureus (S10-2, S10-3, S12-2, and S13-2) isolated from 90 bulgogi samples. The S. aureus enterotoxin H gene (seh) was found in all the strains, while the S. aureus enterotoxin A gene (sea) was found only in 3 of the 4 strains. The S10-2 strain expressed a combination of enterotoxin genes - seg, seh, sei, sej, selm, and seln. The strains S10-2 and S13-2 were resistant to ampicillin and penicillin G, and all the isolated strains were resistant to tetracycline. The S10-2 strain was the only mecA-positive strain; it was also resistant to β-lactam antibiotics. Thus, genes encoding enterotoxin as well as those conferring antibiotic resistance were identified in the S. aureus strains isolated from pork bulgogi. These results represents the potential occurrence of MRSA in pork bulgogi, and the need for a monitoring system for pork bulgogi in order to prevent an outbreak of staphylococcal food poisoning.

DNA microarray-based characterization and antimicrobial resistance phenotypes of clinical MRSA strains from animal hosts

  • Schmitt, Sarah;Stephan, Roger;Huebschke, Ella;Schaefle, Daniel;Merz, Axel;Johler, Sophia
    • Journal of Veterinary Science
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    • v.21 no.4
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    • pp.54.1-54.11
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    • 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.

Rapid Detection of Methicillin Resistant Staphylococcus aureus Based on Surface Enhanced Raman Scattering

  • Han, Dae Jong;Kim, Hyuncheol
    • Korean Journal of Clinical Laboratory Science
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    • v.46 no.4
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    • pp.136-139
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    • 2014
  • Methicillin-resistant Staphylococcus aureus (MRSA) is one of the severe nosocomial infectious agents. The traditional diagnostic methods including biochemical test, antibiotic susceptibility test and PCR amplification are time consuming and require much work. The Surface enhanced Raman spectroscopy (SERS) biosensor is a rapid and powerful tool for analyzing the chemical composition within a single living cell. To identify the biochemical and genetic characterization of clinical MRSA, all isolates from patients were performed with VITEK2 gram positive (GP) bacterial identification and Antibiotic Susceptibility Testing (AST). Virulence genes of MRSA also were identified by DNA based PCR using specific primers. All isolates, which were placed on a gold coated nanochip, were analyzed by a confocal Raman microscopy system. All isolates were identified as S. aureus by biochemical tests. MRSA, which exhibited antibiotic resistance, demonstrated to be positive gene expression of both femA and mecA. Furthermore, Raman shift of S. aureus and MRSA (n=20) was perfectly distinguished by a confocal Raman microscopy system. This novel technique explained that a SERS based confocal Raman microscopy system can selectively isolate MRSA from non-MRSA. The study recommends the SERS technique as a rapid and sensitive method to detect antibiotic resistant S. aureus in a single cell level.

Antimicrobial resistance of Staphylococcus aureus isolated from domestic and imported raw meat in Korea

  • Heo, Hee Jin;Ku, Bok Kyung;Bae, Dong Hwa;Park, Cheong Kyu;Lee, Young Ju
    • 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.

Transmission Aspect of Methicillin-resistant Staphylococcus aureus in the Neurosurgical Intensive Care Unit by Analysing Genotype (유전자형 분석에 의한 신경외과 중환자실의 메티실린 내성 황색포도알균의 전파양상)

  • Kim, Yun-Kyung;Hong, Hae-Sook
    • Journal of Korean Academy of Nursing
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    • v.37 no.6
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    • pp.976-985
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    • 2007
  • Purpose: The aim of this study was to identify the present situation of hospital infection and route of infection by clarifying the transmission aspect of methicillin-resistant Staphylococcus aureus(MRSA) in a Neurosurgical Intensive Care Unit by analysing genotype. Methods: MRSA was cultured from twenty five patients with a tracheostomy, twenty five health care workers, and environments in the Neurosurgical Intensive Care Unit of one hospital in D city. Data was collected from December 21, 2004 to November 5, 2005. MRSA isolates representing each genotype were analyzed by spaA typing and a multiplex PCR method capable of identifying the structural type of the staphylococcal cassette chromosome mec(SCCmec) carried by the bacteria. Results: As the same genotype and gene sequence were found among health care workers, patients, and environments, it was assumed that there was cross transmission among them. Conclusion: This study suggests that first, as the hospital infection by MRSA between health care workers and patients in the Neurosurgical Intensive Care Unit was due to result of cross transmission and the relevance of transmission between them was verified, it is necessary to take preventive measures and conduct education. Secondly, development of nursing interventions and study of infection are needed. Thirdly, consistent investment in prevention against hospital infections and environmental renovation is needed.

Ambient Air Waste Sorting Facilities Could Be a Source of Antibiotic Resistant Bacteria

  • Calheiros, Ana;Santos, Joana;Ramos, Carla;Vasconcelos, Marta;Fernandes, Paulo
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.367-373
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    • 2021
  • The antimicrobial resistance of Staphylococcus spp. and Gram negative strains present in air samples from waste sorting facilities was assessed. Phenotypic studies have revealed a high percentage of strains of Staphylococcus spp. resistant to methicillin. Genotypically and by RT-PCR, it was found that the mecA gene usually associated with methicillin resistance was present in 8% of the Staphylococcus strains isolated. About 30% of the Gram negative strains from the same samples also displayed resistance to meropenem and 79% of these were resistant to multiple antibiotics from different classes, namely cephalosporins and β-lactams. The results suggest that in professional activities with high levels of exposure to biological agents, the quantification and identification of the microbial flora in the work environment, with the determination of the presence of potential agents displaying multi-resistances is of relevance to the risk assessment. The personal protection of workers is particularly important relevance in these cases, since many of the strains that exhibit multi-resistance are potential opportunistic agents.

Pathogenic Characteristics and Antibiotic Resistance of Bacterial Isolates from Farmstead Cheeses

  • Jang, Kyeonga;Lee, Jeeyeon;Lee, Heeyoung;Kim, Sejeong;Ha, Jimyeong;Choi, Yukyung;Oh, Hyemin;Yoon, Yohan;Lee, Soomin
    • Food Science of Animal Resources
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    • v.38 no.1
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    • pp.203-208
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    • 2018
  • The objective of this study was to investigate the pathogenicity and antimicrobial resistance of foodborne pathogens isolated from farmstead cheeses. Twenty-seven isolates, including 18 Bacillus cereus, two Escherichia coli, and seven Staphylococcus aureus, were subjected to polymerase chain reaction (PCR) to detect virulence genes and toxin genes, and the antibiotic resistances of the isolates were determined. All E. coli isolates were determined by PCR to be non-pathogenic. Among the 18 B. cereus isolates, 17 isolates (94.4%) were diarrheal type, as indicated by the presence of nheA, entFM, hbIC, cytK and bceT genes, and one isolate (5.6%) was emetic type, based on the presence of the CER gene. Among the seven S. aureus isolates, three (42.9%) had the mecA gene, which is related to methicillin-resistance. Most B. cereus isolates (94.7%) showed antibiotic resistance to oxacillin and penicillin G, and some strains also showed resistance to ampicillin (26.3%), erythromycin (5.3%), tetracycline (10.5%), and vancomycin (5.3%). These results indicate that microbial food safety measures for farmstead cheese must be implemented in Korea because antibiotic resistant foodborne pathogens, with resistance even to vancomycin, harboring virulence genes were found to be present in the final products of farmstead cheese.