scholarly journals Comparison of Susceptibility Testing Methods withmecA Gene Analysis for Determining Oxacillin (Methicillin) Resistance in Clinical Isolates of Staphylococcus aureus and Coagulase-Negative Staphylococcus spp.

1999 ◽  
Vol 37 (9) ◽  
pp. 2952-2961 ◽  
Author(s):  
P. Kohner ◽  
J. Uhl ◽  
C. Kolbert ◽  
D. Persing ◽  
F. Cockerill
Keyword(s):  
Staphylococcus Aureus ◽  
Susceptibility Testing ◽  
Methicillin Resistance ◽  
Ambient Air ◽  
Coagulase Negative Staphylococcus ◽  
Testing Methods ◽  
Oxacillin Resistance ◽  
Mueller Hinton ◽  
Agar Dilution ◽  
Staphylococcus Spp

Ninety-nine clinical staphylococcal isolates (58 coagulase-negativeStaphylococcus spp. [CoNS] and 41 Staphylococcus aureus isolates) were evaluated for susceptibility to oxacillin. The following susceptibility testing methods, media, and incubation conditions were studied: agar dilution by using Mueller-Hinton (MH) medium (Difco) supplemented with either 0, 2, or 4% NaCl and incubation at 30 or 35°C in ambient air for 24 or 48 h; disk diffusion by using commercially prepared MH medium (Difco) and MH II agar (BBL) and incubation at 35°C in ambient air for 24 or 48 h; and agar screen (spot or swab inoculation) by using commercially prepared agar (Remel) or MH agar (Difco) prepared in-house, each containing 4% NaCl and 6 μg of oxacillin/ml (0.6-μg/ml oxacillin was also studied with MH agar prepared in-house for the agar swab method and CoNS isolates) and incubation at 35°C in ambient air for 24 or 48 h for swab inoculation and at 30 or 35°C in ambient air for 24 or 48 h for spot inoculation. The results for these methods were compared to the results for mecA gene detection by a PCR method. Given the ability to support growth and the results for susceptibility testing (the breakpoint for susceptible isolates was ≤2 μg/ml), the best methods for CoNS isolates were (i) agar dilution by using MH medium supplemented with 4% NaCl and incubation at 35°C for 48 h (no growth failures were noted, and sensitivity was 97.6%) and (ii) agar screen (swab inoculation) by using MH medium prepared in-house supplemented with 4% NaCl and containing 0.6 μg oxacillin/ml and incubation at 35°C for 48 h (one isolate that did not carry the mecA gene did not grow, and the sensitivity was 100%). All but one (agar dilution without added NaCl and incubation at 30°C for 48 h) of the methods tested revealed all oxacillin-resistant S. aureus isolates, and no growth failures occurred with any method. If the breakpoint for susceptibility was lowered to ≤1 μg/ml for agar dilution methods, more CoNS isolates with oxacillin resistance related to the mecA gene were detected when 0 or 2% NaCl agar supplementation was used. Only one CoNS isolate with mecA gene-associated resistance was not detected by using agar dilution and MH medium supplemented with 4% NaCl with incubation for 48 h. When the breakpoint for susceptibility was decreased 10-fold (from 6.0 to 0.6 μg of oxacillin per ml) for the agar swab screen method, fully 100% of the CoNS isolates that carried the mecA gene were identified.

scholarly journals Some coagulase negative Staphylococcus spp. isolated from buffalo can be misidentified as Staphylococcus aureus by phenotypic and Sa442 PCR methods

2018 ◽  
Vol 11 (1) ◽  
Author(s):  
Camila C. de Almeida ◽  
Lucas J. L. Pizauro ◽  
Glenn A. Soltes ◽  
Durda Slavic ◽  
Fernando A. de Ávila ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Coagulase Negative Staphylococcus ◽  
Staphylococcus Spp

scholarly journals Variation in erythromycin and clindamycin susceptibilities of Streptococcus pneumoniae by four test methods.

1997 ◽  
Vol 41 (1) ◽  
pp. 129-134 ◽  
Author(s):  
E L Fasola ◽  
S Bajaksouzian ◽  
P C Appelbaum ◽  
M R Jacobs
Keyword(s):  
Streptococcus Pneumoniae ◽  
Susceptibility Testing ◽  
Ambient Air ◽  
Test Methods ◽  
Disk Diffusion ◽  
Broth Microdilution ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
Agar Dilution ◽  
Resistant Strains

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.

scholarly journals Susceptibility of Multiresistant Gram-Negative Bacteria to Fosfomycin and Performance of Different Susceptibility Testing Methods

2013 ◽  
Vol 58 (3) ◽  
pp. 1763-1767 ◽  
Author(s):  
L. V. Perdigão-Neto ◽  
M. S. Oliveira ◽  
C. F. Rizek ◽  
C. M. D. M. Carrilho ◽  
S. F. Costa ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Klebsiella Pneumoniae ◽  
Susceptibility Testing ◽  
Gram Negative Bacteria ◽  
Testing Methods ◽  
Gram Negative ◽  
Content Type ◽  
Agar Dilution ◽  
And Performance ◽  
Systemic Infections

ABSTRACTFosfomycin may be a treatment option for multiresistant Gram-negative bacteria. This study compared susceptibility methods using 94 multiresistant clinical isolates. With agar dilution (AD), susceptibilities were 81%, 7%, 96%, and 100% (CLSI) and 0%, 0%, 96%, and 30% (EUCAST), respectively, forAcinetobacter baumannii,Pseudomonas aeruginosa,Klebsiella pneumoniae, andEnterobacterspp. Categorical agreement between Etest and AD forEnterobacteriaceaeandA. baumanniiwas ≥80%. Disk diffusion was adequate only forEnterobacter. CLSI criteria for urine may be adequate for systemic infections.

scholarly journals OptimizedIn VitroAntibiotic Susceptibility Testing Method for Small-Colony Variant Staphylococcus aureus

2016 ◽  
Vol 60 (3) ◽  
pp. 1725-1735 ◽  
Author(s):  
Mimi R. Precit ◽  
Daniel J. Wolter ◽  
Adam Griffith ◽  
Julia Emerson ◽  
Jane L. Burns ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Susceptibility Testing ◽  
Brain Heart Infusion ◽  
Chronic Infections ◽  
Poor Growth ◽  
Content Type ◽  
Treatment History ◽  
Mueller Hinton ◽  
Mueller Hinton Agar ◽  
Small Colony

Staphylococcus aureussmall-colony variants (SCVs) emerge frequently during chronic infections and are often associated with worse disease outcomes. There are no standardized methods for SCV antibiotic susceptibility testing (AST) due to poor growth and reversion to normal-colony (NC) phenotypes on standard media. We sought to identify reproducible methods for AST ofS. aureusSCVs and to determine whether SCV susceptibilities can be predicted on the basis of treatment history, SCV biochemical type (auxotrophy), or the susceptibilities of isogenic NC coisolates. We tested the growth and stability of SCV isolates on 11 agar media, selecting for AST 2 media that yielded optimal SCV growth and the lowest rates of reversion to NC phenotypes. We then performed disk diffusion AST on 86S. aureusSCVs and 28 isogenic NCs and Etest for a subset of 26 SCVs and 24 isogenic NCs. Growth and reversion were optimal on brain heart infusion agar and Mueller-Hinton agar supplemented with compounds for which most clinical SCVs are auxotrophic: hemin, menadione, and thymidine. SCVs were typically nonsusceptible to either trimethoprim-sulfamethoxazole or aminoglycosides, in accordance with the auxotrophy type. In contrast, SCVs were variably nonsusceptible to fluoroquinolones, macrolides, lincosamides, fusidic acid, and rifampin;mecA-positive SCVs were invariably resistant to cefoxitin. All isolates (both SCVs and NCs) were susceptible to quinupristin-dalfopristin, vancomycin, minocycline, linezolid, chloramphenicol, and tigecycline. Analysis of SCV auxotrophy type, isogenic NC antibiograms, and antibiotic treatment history had limited utility in predicting SCV susceptibilities. With clinical correlation, this AST method and these results may prove useful in directing treatment for SCV infections.

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