scholarly journals Evaluation of different phenotypic methods for the detection of methicillin resistant Staphylococcus aureus and antimicrobial susceptibility pattern of MRSA

2017 ◽  
Vol 4 (9) ◽  
pp. 3297 ◽  
Author(s):  
Sheetal Sharma ◽  
Preeti Srivastava ◽  
Anjali Kulshrestha ◽  
Ameer Abbas
Keyword(s):  
Staphylococcus Aureus ◽  
Sensitivity And Specificity ◽  
Antimicrobial Susceptibility ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Phenotypic Methods ◽  
Cefoxitin Disk ◽  
Cefoxitin Disk Diffusion

Background: Rapid and accurate detection of methicillin resistant Staphylococcus aureus (MRSA) is an important role of clinical microbiology laboratories to avoid treatment failure. The aim of this study was to compare conventional methods against the cefoxitin disc diffusion method to determine the best phenotypic method. Methods: Study was carried out in the Department of Microbiology, National Institute of Medical Sciences & Research, Jaipur (India), between July 2016 – December 2016. The methods included were Oxacillin E-test MIC, Oxacillin screen agar, Oxacillin disk diffusion, Cefoxitin disk diffusion and CHROMagar- MRSA methods. Antimicrobial susceptibility performed as per CLSI guidelines.Results: Out of 142 isolates of S. aureus, fifty three (37.32%) strains of MRSA were isolated from clinical specimen. E-MIC test was selected as gold standard method. The sensitivity and specificity of Oxacillin screen agar and CHROMagar-MRSA were same 98.07% and 97.80%, respectively. The sensitivity and specificity of oxacillin disk diffusion were 94.23% and 98.89%. Fifty three strains of S. aureus were MRSA by cefoxitin disk diffusion method and Oxacillin Ezy MIC test. The sensitivity and specificity of cefoxitin disk diffusion method and Oxacillin Ezy MIC method was 100% and 100% respectively. All isolates including MRSA were susceptible to Vancomycin and Linezolid. Conclusions: All phenotypic methods had high sensitivity and specificity for detection of MRSA. However, cefoxitin disk diffusion method in comparison to other methods had higher sensitivity and specificity. 

scholarly journals Multicenter Evaluation of a Modified Cefoxitin Disk Diffusion Method and PBP2a Testing To PredictmecA-Mediated Oxacillin Resistance in Atypical Staphylococcus aureus

2016 ◽  
Vol 55 (2) ◽  
pp. 485-494 ◽  
Author(s):  
Shelley A. Miller ◽  
James Karichu ◽  
Peggy Kohner ◽  
Nicolynn Cole ◽  
Janet A. Hindler ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Clinical Laboratory ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Content Type ◽  
Oxacillin Resistance ◽  
Mueller Hinton ◽  
Cefoxitin Disk ◽  
Cefoxitin Disk Diffusion

ABSTRACTPhenotypic variants ofStaphylococcus aureusthat display small colonies, reduced pigmentation, and decreased hemolysis and/or coagulase activity are periodically isolated by the clinical laboratory. Antimicrobial susceptibility testing (AST) of these isolates is complicated, because many do not grow on routine AST media, including Mueller-Hinton agar (MHA) and cation-adjusted Mueller-Hinton broth. This multicenter study evaluated cefoxitin disk diffusion for 37 atypicalS. aureusisolates (156 readings) with MHA supplemented with 5% sheep's blood (BMHA), usingmecAPCR as the reference standard. The correlation of two commercial PBP2a assays withmecAPCR was also assessed. Ten isolates were negative and 27 positive formecA. No major errors for cefoxitin were observed, but 19.5% very major errors (VMEs) were observed at 24 h of incubation, and 17.2% VMEs were observed at 48 h. The proportions of VMEs ranged from 14.7 to 23.0% at 24 h, and from 13.3 to 17.6% at 48 h, across three testing laboratories. PBP2a tests were performed from growth on BMHA and blood agar plates (BAP), with and without cefoxitin disk induction. The Alere PBP2a SA culture colony test sensitivities formecAwere 90.0% with uninduced growth and 97.4% with induced growth from BMHA. On BAP, sensitivity was 96.0% with induced growth. The sensitivities of the Oxoid PBP2′ latex agglutination test were 85.7% with uninduced growth and 93.9% with induced growth from BMHA and 95.9% with induced growth on BAP. On the basis of these data, we recommend that laboratories perform onlymecAPCR and/or PBP2a tests when requested to perform AST on atypical isolates ofS. aureus.

scholarly journals Detection of Oxacillin Resistance in Staphylococcus aureus Isolated from the Neonatal and Pediatric Units of a Brazilian Teaching Hospital

2009 ◽  
Vol 3 ◽  
pp. CMPed.S2085 ◽  
Author(s):  
Valéria Cataneli Pereira ◽  
André Martins ◽  
Lígia Maria Suppo de Souza Rugolo ◽  
Maria de Lourdes Ribeiro de Souza da Cunha
Keyword(s):  
Staphylococcus Aureus ◽  
Screening Test ◽  
Blood Cultures ◽  
Neonatal Unit ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Fluid Sample ◽  
Disk Diffusion Method ◽  
Cefoxitin Disk ◽  
Cefoxitin Disk Diffusion

Objective To determine, by phenotypic and genotypic methods, oxacillin susceptibility in Staphylococcus aureus strains isolated from pediatric and neonatal intensive care unit patients seen at the University Hospital of the Botucatu School of Medicine. Methods A total of 100 S. aureus strains isolated from the following materials were studied: 25 blood cultures, 21 secretions, 12 catheters, 3 cannulae and one chest drain from 62 patients in the neonatal unit, and 36 blood cultures, one pleural fluid sample and one peritoneal fluid sample from 38 patients in the pediatric unit. Resistance of the S. aureus isolates to oxacillin was evaluated by the disk diffusion method with oxacillin (1 μg) and cefoxitin (30 μg), agar screening test using Mueller-Hinton agar supplemented with 6 μg/ml oxacillin and 4% NaCl, and detection of the mecA gene by PCR. In addition, the isolates were tested for β-lactamase production using disks impregnated with Nitrocefin and hyperproduction of β-lactamase using amoxicillin (20 μg) and clavulanic acid (10 μg) disks. Results Among the 100 S. aureus strains included in the study, 18.0% were resistant to oxacillin, with 16.1% MRSA being detected in the neonatal unit and 21.0% in the pediatric unit. The oxacillin (1 μg) and cefoxitin (30 μg) disk diffusion methods presented 94.4% and 100% sensitivity, respectively, and 98.8% specificity. The screening test showed 100% sensitivity and 98.8% specificity. All isolates produced β-lactamase and one of these strains was considered to be a hyperproducer. Conclusions The 30 μg cefoxitin disk diffusion method presented the best result when compared to the 1 μg oxacillin disk. The sensitivity of the agar screening test was similar to that of the cefoxitin disk diffusion method and higher than that of the oxacillin disk diffusion method. We observed variations in the percentage of oxacillin-resistant isolates during the study period, with a decline over the last years which might be related to improved nosocomial infection control and the rational use of antibiotics.

Evaluation of antibiotic resistance pattern and mecA gene frequency in methicillin-resistant Staphylococcus aureus strains collected from clinical specimens in Marand hospital and treatment centers, Iran

2021 ◽  
Author(s):  
Abolfazl Jafari-Sales ◽  
Zahra Sadeghi Deylamdeh ◽  
Afsoon Shariat
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Meca Gene ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Methicillin Resistant ◽  
Medical Centers ◽  
Wide Range

Introduction: Staphylococcus aureus causes a wide range of infections and as a multivalent pathogen is one of the causative agents of nosocomial and community infections. Therefore, the aim of this study was to identify and determine the pattern of antibiotic resistance of methicillin-resistant Staphylococcus aureus (MRSA) isolates from patients in hospitals and medical centers in Marand city and also to evaluate the presence of mecA gene. Materials and Methods: In this cross-sectional descriptive study, 385 samples of S. aureus were collected from different clinical samples of patients in hospitals and medical centers of Marand city. S. aureus was identified using standard biochemical methods.  Methicillin resistance was determined by disk diffusion method in the presence of oxacillin and cefoxitin. The pattern of antibiotic resistance of the strains was determined by disk diffusion method and according to CLSI recommendation and also PCR method was used to evaluate the frequency of MecA gene. Results: In the present study, out of 385 samples of S. aureus, 215 (55.84%) samples were methicillin resistant. PCR results for mecA gene showed that 110 samples had mecA gene.  The highest antibiotic resistance was observed against penicillin (100%) and erythromycin (83.63%). Most MRSA were isolated from urine and wound samples. Conclusion: The results of this study indicate the prevalence of methicillin-resistant species and also the increase in antibiotic resistance of MRSA to various antibiotics.  Therefore, in order to prevent increased resistance to other antibiotics, it is recommended to avoid inappropriate use of antibiotics.

scholarly journals Genetic Characterization of Methicillin-Resistant Staphylococcus aureus Isolates from Human Bloodstream Infections: Detection of MLSB Resistance

Antibiotics ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 375
Author(s):  
Vanessa Silva ◽  
Sara Hermenegildo ◽  
Catarina Ferreira ◽  
Célia M. Manaia ◽  
Rosa Capita ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Agents ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Bloodstream Infections ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Accessory Gene ◽  
Spa Typing ◽  
Disk Diffusion Method ◽  
Methicillin Resistant

In this study we aimed to characterize antimicrobial resistance in methicillin-resistant Staphylococcus aureus (MRSA) isolated from bloodstream infections as well as the associated genetic lineages of the isolates. Sixteen MRSA isolates were recovered from bacteremia samples from inpatients between 2016 and 2019. The antimicrobial susceptibility of these isolates was tested by the Kirby–Bauer disk diffusion method against 14 antimicrobial agents. To determine the macrolide–lincosamide–streptogramin B (MLSB) resistance phenotype of the isolates, erythromycin-resistant isolates were assessed by double-disk diffusion (D-test). The resistance and virulence genes were screened by polymerase chain reaction (PCR). All isolates were characterized by multilocus sequence typing (MLST), spa typing, staphylococcal chromosomal cassette mec (SCCmec) typing, and accessory gene regulator (agr) typing. Isolates showed resistance to cefoxitin, penicillin, ciprofloxacin, erythromycin, fusidic acid, clindamycin, and aminoglycosides, confirmed by the presence of the blaZ, ermA, ermC, mphC, msrA/B, aac(6’)-Ie-aph(2’’)-Ia, and ant(4’)-Ia genes. Three isolates were Panton–Valentine-leukocidin-positive. Most strains (n = 12) presented an inducible MLSB phenotype. The isolates were ascribed to eight spa-types (t747, t002, t020, t1084, t008, t10682, t18526, and t1370) and four MLSTs (ST22, ST5, ST105, and ST8). Overall, most (n = 12) MRSA isolates had a multidrug-resistance profile with inducible MLSB phenotypes and belonged to epidemic MRSA clones.

scholarly journals Contamination and Antimicrobial Susceptibility Testing of Staphylococcus aureus Isolated from Pork in Fresh Markets, Nongchok District, Thailand

2021 ◽  
Vol 2021 ◽  
pp. 1-3
Author(s):  
Doungjit Kanungpean ◽  
Shinji Takai ◽  
Tsutomu Kakuda
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Severe Infection ◽  
Antimicrobial Susceptibility Testing ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Meat Markets ◽  
Pork Samples

We surveyed Staphylococcus aureus contamination in 110 pork samples from 12 fresh meat markets in Nongchok district, Bangkok, Thailand, and performed antimicrobial susceptibility testing with the disk diffusion method. The prevalence of S. aureus was 28.18%, and 52 strains were isolated. Antimicrobial susceptibility testing using the disk diffusion method revealed that 80.77% of the isolates were resistant to tetracycline and 76.92% to ampicillin. All strains were 100% susceptible to cloxacillin, cefoxitin, gentamicin, and cefazolin. The high percentage of antibiotic resistance to tetracycline and ampicillin was attributed to their use in treating infections in farmed animals and their addition to animal food for disease prevention. Interestingly, the present study revealed the intermediate resistance of S. aureus (13.46% of S. aureus-positive pork samples) to vancomycin which is a common medicine for treating severe infection in humans, suggesting that the trend of resistance might increase and becoming a serious problem of public health for both humans and animals.

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