Evaluation of Identification and Susceptibility for Candida Spp. Isolated Directly from Positive Blood Culture Bottles

2021 ◽  
Vol 2021 ◽  
pp. 1-5
Ândrea Celestino de Souza ◽  
Luciano Z. Goldani ◽  
Eliane Würdig Roesch ◽  
Larissa Lutz ◽  
Patricia Orlandi Barth ◽  

Determination of the susceptibility profile of isolates of Candida from blood culture bottles is extremely important for correctly guiding patient pharmacotherapy. The aim of this study was to compare the results of analysis of Candida isolated directly from blood culture bottles by the VITEK MS MALDI-TOF identification system and the fluconazole disk diffusion assay with those of standard identification methods. Testing directly from the bottle allowed results 24 to 48 hours quicker than the standard method. There was a categorical agreement of 51.64% (47 of 91 samples) between the results of analysis directly from the bottle and analysis by the standard method. Regarding species identification, there was 96.15% agreement for Candida parapsilosis (25 of 26 samples). Categorical agreement between the rapid and standard disk diffusion methods was 95%, and the agreement between the rapid disk diffusion method and the broth microdilution method was 97%. Only minor errors in the rapid method were observed: 3 (5%) in the standard disk diffusion method and 2 (3%) in the broth microdilution method. Our study concluded that the rapid disk diffusion method for fluconazole is a fast, easy, reproducible, and consistent method. Its timely implementation for testing antifungal agents in the clinical microbiology laboratory can help reduce profile release times, thus helping to determine the most appropriate antifungal treatment.

2006 ◽  
Vol 50 (3) ◽  
pp. 1108-1111 ◽  
E. López-Oviedo ◽  
A. I. Aller ◽  
C. Martín ◽  
C. Castro ◽  
M. Ramirez ◽  

ABSTRACT The disk diffusion method was evaluated for determining posaconazole susceptibility against 78 strains of molds using two culture media in comparison with the CLSI (Clinical Laboratory Standards Institute) broth microdilution method (M38-A). A significant correlation between disk diffusion and microdilution methods was observed with both culture media.

2006 ◽  
Vol 50 (7) ◽  
pp. 2330-2336 ◽  
Helio S. Sader ◽  
Thomas R. Fritsche ◽  
Ronald N. Jones

ABSTRACT A total of 207 Staphylococcus aureus strains, including 105 well-characterized strains with decreased susceptibility to vancomycin (17 vancomycin-intermediate S. aureus [VISA] and 88 heteroresistant VISA [hVISA] strains) and 102 wild-type methicillin-resistant S. aureus (MRSA-WT) strains were tested by reference/standardized broth microdilution and disk diffusion methods, as well as by Etest (AB BIODISK, Solna, Sweden), against daptomycin and vancomycin. The lowest concentration of antimicrobial agent that killed ≥99.9% of the initial inoculum was defined as the minimum bactericidal concentration (MBC) endpoint, and time-kill curves were performed in selected strains to further evaluate bactericidal activity. All MRSA-WT and hVISA strains were inhibited by ≤1 μg/ml of daptomycin, while the VISA strains showed slightly higher daptomycin MICs (range, 0.5 to 4 μg/ml). All daptomycin MBC results were at the MIC or twofold higher. In contrast, 14.7% of MRSA-WT, 69.3% of hVISA, and all VISA strains showed a vancomycin MBC/MIC ratio of ≥32 or an MBC of ≥16 μg/ml (tolerant). The correlation coefficients between broth microdilution and disk diffusion method results were low for daptomycin (0.07) and vancomycin (0.11). Eight (3.8%) strains (all hVISA or VISA) were “nonsusceptible” to daptomycin by broth microdilution methods but susceptible by the disk diffusion method. For vancomycin, 35 (16.9%) strains were nonsusceptible by broth microdilution methods but susceptible by disk diffusion methods. In conclusion, daptomycin was highly bactericidal against S. aureus strains, and its bactericidal activity was not affected by decreased susceptibility to vancomycin. In contrast, many (one in seven) contemporary MRSA-WT, the majority of hVISA, and all VISA strains showed vancomycin MBC/MIC ratios consistent with tolerance, a predictor of poor clinical response. Disk diffusion tests generally failed to detect strains categorized as nonsusceptible to daptomycin or vancomycin by the reference broth microdilution method or Etest, and reassessment of breakpoints should be immediately attempted for MIC methods suggested as the test of choice.

2012 ◽  
Vol 66 (3-4) ◽  
pp. 199-210
Jelena Asanin ◽  
Ksenija Aksentijevic ◽  
Milenko Zutic ◽  
Vera Katic ◽  
Dejan Krnjaic ◽  

The objective of this work was to identify isolated Staphylococcus species and to investigate their sensitivity to some antibacterial drugs. The material used for these investigations were Staphylococcus isolates originating from milk samples. A total of 25 strains of Staphylococcus isolates were examined, including 24 from milk samples from cows with mastitis, and one strain was isolated from a milk sample from a cow following treatment for mastitis. For primary identification, catalase and oxidase tests were used, as well as the free coagulase test. Following the preliminary tests, the isolated strains were identified using commercial systems ID32 STAPH (bioM?rieux, France) and the BBL Crystal Gram-Positive ID Kit (Becton Dickinson, USA) according to the enclosed instructions. The Staphylococcus isolates were examined for sensitivity to the following: oxacillin, penicillin, cefoxitin, gentamicin, erythromycin, chloramphenicol, tetracycline, ciprofloxacin, sulfametoxazol/trimetoprim, and vacomycin using the disk diffusion method and the broth microdilution method as recommended by the Clinical and Laboratory Strandards Institute - CLSI(2003), and the results were interpreted according to CLSI recommendations from 2008 and 2010. Antibiogram disks manufactured by Becton Dickinson (USA) were used, and the broth microdilution method was applied using pure antibiotic substances from different manufacturers: erythromycin, chloramphenicol, cefoxitin, gentamicin, oxacillin, tetracycline (Sigma Aldrich, USA), sulfametoxazol (Fluka, USA), penicillin (Calbiochem, Germany), vancomycin (Abbott laboratories, USA), ciprofloxacin and trimetoprim (Zdravlje A.D., Serbia). All 25 strains were catalase positive and oxidase negative. Of the 25 strains, 19 were coagulase positive and 6 were coagulase negative.With the implementation of the disk diffusion method on 19 strains of S. aureus, 17 were established to be resistant to penicillin (89.5%), and 2 strains to gentamicin (10.5%). The investigation of 3 strains of S. xylosus using the disk diffusion method showed that one strain was resistant to tetracycline (33.3%) and to oxacillin (33.3%), while another strain was found to be resistant to penicillin (33.3%). The third strain of S. xylosus was sensitive to all the examined antibiotics. Two strains of S. simulans and one strain of S. haemolyticus were not found to be resistant to any of the examined antibiotics using the disk diffusion method. The implementation of the broth microdilution method yielded in 13 strains of S. aureus resistance to penicillin (68.4%) with MIC values from 0.5 to 4 ?g/m, in 2 strains to gentamicin (10.5%) with MIC values of 32 ?g/ml, and intermediary sensitivity to chloramphenicol was established in 9 strains of S. aureus (47.4%) with MIC values of 16 ?g/ml and to vancomycin in 1 strain of S. aureus (5.3%) with MIC values of 4 ?g/ml.

Natalie S. Nunnally ◽  
Tajah Damm ◽  
Shawn R. Lockhart ◽  
Elizabeth L. Berkow

We evaluated the CLSI M44ed3E disk diffusion method in comparison with the CLSI M27ed4 broth microdilution method for caspofungin and fluconazole and the Etest method for amphotericin B to categorize susceptibility of 347 clinical isolates of Candida auris. Utilizing the zone diameter cutoffs established here we observed the overall categorial agreement between the two methods. For caspofungin, concordant results were observed for 98% of isolates with <1% very major and 1% major errors. For fluconazole, concordant results were observed for 91% of isolates with 1% very major and 8% major errors. For amphotericin B, concordant results were observed for 74% of isolates with <1% very major errors and 25% major errors. The disk diffusion approach provides an accurate method for determining the susceptibility of C. auris for caspofungin and fluconazole, and for identification of at least 75% of amphotericin B-susceptible isolates.

1997 ◽  
Vol 41 (1) ◽  
pp. 129-134 ◽  
E L Fasola ◽  
S Bajaksouzian ◽  
P C Appelbaum ◽  
M R Jacobs

Susceptibilities of 124 strains of Streptococcus pneumoniae to erythromycin and clindamycin were determined by the National Committee for the Clinical Laboratory Standards (NCCLS) broth microdilution method, with incubation for 20 to 24 h in ambient air and with modifications of this method by incubation for up to 48 h in air and CO2. Strains were also tested by agar dilution, E-test, and disk diffusion; good correlation was obtained with these methods, with clear separation into bimodal populations of susceptible and resistant stains. The broth microdilution method, however, using incubation in air for 24 h (NCCLS method), misclassified 4 of 92 erythromycin-resistant strains (1 as susceptible and 3 as intermediate) and 25 of 58 clindamycin-resistant strains (all as susceptible). With the exception of one strain with clindamycin, susceptible and resistant strains were correctly classified by the microdilution method with incubation in CO2 for 24 h or in ambient air for 48 h. Disk diffusion, agar dilution, and E-test methods with incubation in 5% CO2 are therefore reliable methods for susceptibility testing of pneumococci against these agents. However, the NCCLS microdilution method, which specifies incubation for 20 to 24 h in ambient air, produced significant very major errors (43%) clindamycin. Modification of the microdilution method by incubation in 5% CO2 or by extension of incubation time in ambient air to 48 h corrected these errors. Disk diffusion, however, was shown to be a simple, convenient, and reliable method for susceptibility testing of pneumococci to erythromycin and clindamycin and is suggested as the method of choice for these agents.

2008 ◽  
Vol 52 (No. 4) ◽  
pp. 169-174
M. Holasova ◽  
R. Karpiskova ◽  
S. Karpiskova ◽  
V. Babak ◽  
J. Schlegelova

With a microdilution method, using the commercial diagnostic test Sensititre Susceptibility Plates for Campylobacter MIC (Trek Diagnostic Systems, Cleveland, OH, USA), disk diffusion and agar dilution method, resistance to six antimicrobial agents were examined in a reference strain <i>Campylobacter jejuni</i> ATCC 33560 and 73 thermo-tolerant isolates of <i>Campylobacter</i> spp. For the microdilution method and all tested antimicrobial agents, our determined values of microbiological breakpoints of resistant strains were suggested as the minimum inhibitory concentration (MIC<sub>R</sub>) for ciprofloxacin &ge; 0.5, erythromycin &ge; 4, gentamicin &ge; 4, nalidixic acid &ge; 32 and tetracycline &ge; 4 &mu;g/ml. On the basis of our study results, strains resistant to clindamycin were MIC<sub>R</sub> &ge; 2 &mu;g/ml for the dilution methods and a zone diameter R ≤ 16 mm for the disk diffusion method. Comparison of the results of the resistance examination, a microdilution method and disk diffusion method with the reference agar dilution method, showed that all compared methods yielded identical results with the exception of the resistance determination in erythromycin and nalidixic acid. The errors were mostly the result of the interpretation criteria for MIC<sub>R</sub> of agar dilution method and different conditions of cultivation used. However, the compared methods, provide results comparable with the reference method having greater convenience of measurement.

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