• Biomedical Science Letters
• /
• v.25 no.1
• /
• 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.

• Food Science of Animal Resources
• /
• v.43 no.5
• /
• pp.792-804
• /
• 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.

• Journal of Veterinary Clinics
• /
• v.31 no.3
• /
• pp.194-198
• /
• 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.

• Korean Journal of Clinical Laboratory Science
• /
• v.49 no.4
• /
• pp.420-426
• /
• 2017

Methicillin-resistant Staphylococcus (MRS) can be colonized in various body sites and is more frequently isolated in healthcare associated persons. This study aimed to evaluate the contamination rate of MRS in a high school environment, those living with closed life style. Staphylococcus aureus was isolated from only the hands of 2 students among a sample of 28 students, and S. aureus were susceptible to methicillin antibiotics. Coagulase negative Staphylococci (CoNS) were isolated from the hands of 26 students (26/28, 92.9%), and among them, 14 (53.8%) isolates were methicillin-resistant CoNS (MRCoNS). Among the 14 MRCoNS, S. warneri was the most common (8/14, 57%) and susceptible to most $non-{\beta}-lactam$ antibiotics, such as clindamycin, erythromycin, ciprofloxacin, tetracycline, gentamicin, and vancomycin. In a culture of 31 desks, S. aureus was not isolated but CoNS were isolated from 26 desks (26/31, 83.6%), which did not harbor the mecA gene. The other bacteria isolated from the hands and desks were Micrococcus and Bacillus spp. In conclusion, methicillin-resistant S. aureus was not isolated from the hands and desks of high school students. However, the frequency of MRCoNS harboring mecA gene were high in the hands of high school students. Therefore, to prevent and to control the transfer of infection, intensifying preventive education, such as hand washing, and active surveillance systems, such as an investigation of contamination or carrier rate of resistant bacteria are necessary.

• The Journal of the Korean Society for Microbiology
• /
• v.22 no.3
• /
• pp.283-294
• /
• 1987

One hundred and fifty-seven strains of staphylococci isolated from various clinical specimens and 80 of Gram-negative bacilli from urine of patients with urological diseases were tested for resistance to antimicrobial drugs by microdilution broth method. Among staphylococci, 50 to 89% of the strains were resistant to gentamicin(Gm), kanamycin(Km), erythromycin(Em), nalidixic acid(Na), and tetracycline. Ninety per cent MIC was lowest in ciprofloxacin(Cp), followed by vancomycin(Vc), trimethoprim(Tp), enoxacin(Ex), and norfloxacin(Nf) with the values of two ${\mu}g/ml$ or lower. Twenty-seven strains were resistant to methicillin(MR), with 24 strains of Staphylococcus aureus and 3 of S. epidermidis. All strains of MR S. aureus were resistant to oxacillin, rifampin(Rf), Gm, Km, Em, Na, and Tc, and no strain was resistant to Vc and Tp. Almost all staphylococci isolated from urine were S. epidermidis and sensitive to most drugs tested without MR strain. Among Gram-negative bacilli from urine, Escherichia coli(43 strains) was most frequently isolated, and followed by Klebsiella spp.(11), Proteus spp.(10), Serratia spp.(10), and Pseudomonas aeruginosa(6) in the decreasing order. The majority of E. coli and Serratia spp. were resistant to chloramphenicol(Cm), Tc, streptomycin, sulfisomidine(Su), ampicillin(Ap), Km, and carbenicillin(Cb), and 50 and 90% MICs of these drugs were also high. In Klebsiella spp., 54% or more were resistant to Cm, Su, Ap, cephalothin, and Cb. Proteus spp. were susceptible to most drugs tested, but Pseudomonas were resistant to nearly all drugs tested except Rf, amikacin, and moxalactam(Mx). All Gram-negative bacilli tested were found to be susceptible to Mx. New quinolone carboxylic acid compounds, such as Nf, Ex, and Cp showed very high antimicrobial activities against the majority of organisms tested except Pseudomonas, and 50 and 90% MICs of Nf and Ex were always equal or 2 to 4 times higher than Cp. Organisms multiply resistant to drugs were noted in almost all isolates tested. Twenty-seven strains of staphylococci were multiply resistant to 11 or more drugs, and 6 of Klebsiella spp. to 8 to 11 drugs. The most frequent multiplicity of durg resistance were 7 and 8, 12, and 13 in E. coli, Serratia spp., and Pseudomonas, respectively. No strain was resistant to more than 5 drugs in Proteus spp..

• Proceedings of the Zoological Society Korea Conference
• /
• 1999.10b
• /
• pp.211.2-211
• /
• 1999

No Abstract, See Full Text

• Journal of Dairy Science and Biotechnology
• /
• v.35 no.3
• /
• pp.162-168
• /
• 2017

Mastitis is a common and serious infection of the mammary gland in dairy cattle and has a major economic impact on the production of milk and dairy products. Bacterial mastitis is caused by several pathogens and is most frequently associated with coagulase-negative staphylococci (CNS). Although CNS are typically associated with subclinical or mild mastitis, the importance of CNS has increased as these pathogens have emerged as predominant mastitis-related pathogens in many countries. CNS can cause persistent infections, resulting in increased milk somatic cell counts and thereby affecting milk quality and decreasing milk production. Globally, Staphylococcus chromogenes, S. epidermidis, and S. simulans are the predominant CNS species in dairy cattle mastitis. Antibacterial resistance of CNS varies with species, and most CNS are susceptible to vancomycin and resistant to penicillin and tetracycline. As the most frequently isolated CNS species, some strains of S. chromogenes exhibit phenotypic resistance to ampicillin, erythromycin, oxacillin, penicillin, and tetracycline. Some strains of S. epidermidis and S. haemolyticus are only susceptible to vancomycin and rifampicin. Therefore, more studies are needed to achieve the control and prevention of CNS as environmental pathogens.

• Journal of Chest Surgery
• /
• v.9 no.2
• /
• pp.125-132
• /
• 1976

The author made clinical study of 100 cases of empyema in infancy and childhood that were treated at the Department of Chest Surgery, Busan University Hospital and Busan Children's Charity Hospital, from Jan 1962 to Nov. 1975. 1. In infancy and childhood, 62 cases out of my 100 cases of empyema were caused by .staphylococci and most of recent reports showed a gradual increase in number of staphylococcal empyema. 2. Most frequent lesion predisposing to empyema in infancy and childhood was pneumonia (72%), being remarkable in staphylococcal empyema (85.5%) to that of others. 3. Antibiotics sensitivity test for staphylococci revealed that the erythromycin was most susceptible (85. 5%). 4. The mortality rate was 6% in over all and the author believes that from the point of view of surgical treatment, failure of early continuous drainage on account of multiple thoracentesis for the early stage of empyema, and also early open thoracotomy procedure such as decortication were all the contributing factors to higher mortality in the empyema of infancy and childhood. 5. It may be concluded that the treatment of choice for empyema in infancy and childhood were early and prolonged continuous drainage of pus by closed thoracotomy with caution and administration of more susceptible antibiotics with nutritional support.

• Journal of Microbiology
• /
• v.35 no.2
• /
• pp.103-108
• /
• 1997

To improve the detection of methicillin resistant staphylococci, lowered incubation temperature (30.deg.) and inclusion of sodium chloride in media have been empirically recommended. However, in this study, we found that sodium chloride in Peptone-Yeast Extract-K$\_$2/HPO$\_$4/ (PYK) medium decreased methicillin minimum inhibitory concentrations. Divalent cations were shown to restore the expression of staphylococcal methicillin resistance. However, when it was determined by efficiency of plating, sodium chloride increased methicillin resistance expression on agar medium in which higher divalent cations were contained in the agar medium. The decrease of minimum inhibitory concentrations at 30.deg.C by sodium chloride occurred in Brain Heart Infusion but did not occur in other media investigated. Interestingly, both PYK and Brain Heart Infusion media had peptone, which contain cholic acids having detergent activities. Inclusion of sodium chloride in PYK caused a higher rate of autolysis. Penicillin binding protein 2a that has a low affinity to beta-lactam antibiotics, was highly inducible in methicillin resistant Staphylococcus epidermidis strains. In this study, we found that autolysins that are activated by the sodium chloride decreased the minimum inhibitory concentration at 30.deg.C, and peptidoglycan is weakened due to the presence of methicillin. Peptone in the media may aggravate the fragile cells. However, stabilization due to the presence of divalent cations and production of penicilin binding protein 2a increase the survival of staphylococci.

• International Journal of Oral Biology
• /
• v.40 no.1
• /
• pp.41-50
• /
• 2015

The aim of this study was to identify bacteria isolated from the oral cavities and to determine their antimicrobial susceptibility against eight antibiotics. The bacterial strains were obtained from the Korean Collection for Oral Microbiology (KCOM). The bacteria were identified by comparing 16S rDNA sequences at the species level. The data showed that 77 bacterial strains were predominantly identified as streptococci (49.4%) and staphylococci (14.3%). Minimum inhibitory concentrations (MIC) were determined using a broth dilution assay to test the sensitivity of the bacterial strains. The MIC values of the oral bacterial strains against antibiotics were different. Streptococci were sensitive to clindamycin, cefuroxime axetil, and vancomycin, and they were resistant to tetracycline. Staphylococci also were sensitive to clindamycin, cefuroxime axetil, and vancomycin, and they were resistant to penicillin antibiotics. Gramnegative bacterial strains were sensitive to tetracycline and were resistant to clindamycin. These results suggest that the antimicrobial susceptibility test is necessary in deciding the prescription for antibiotics, to prevent the misuse or abuse of antibiotics.