Comparison of Etest, Vitek and agar dilution for susceptibility testing of colistin

In total, 172 isolates of Enterobacteriaceae, Acinetobacter spp., Pseudomonas aeruginosa and Stenotrophomonas maltophilia were tested for susceptibility to colistin by agar dilution, Etest and the Vitek 2 system. Isolates with a colistin MIC < or =2 mg/L were considered to be susceptible. Fifty-four (31%) Gram-negative isolates were resistant to colistin. Categorical agreement between agar dilution and Etest was 87%, and between agar dilution and Vitek 2 was 82%. Based on the data obtained, the Vitek 2 system was unreliable for detecting colistin resistance, and results obtained by Etest may require confirmation by a standard MIC susceptibility testing method.     

Comparison of PCR detection of mecA with agar dilution and Etest for oxacillin susceptibility testing in clinical isolates of coagulase-negative staphylococci

Oxacillin-resistant staphylococci are heterogeneous in their expression of resistance to beta-lactam antibiotics. Different recommendations regarding screening methods for routine use have been published. In this study, the susceptibility to oxacillin of 232 coagulase-negative staphylococci (CoNS) was determined by agar dilution, Etest and presence of the mecA gene. When an oxacillin resistance breakpoint of > or = 0.5 mg/L was used, the sensitivity and specificity for agar dilution were 97.6% and 100%, and those for Etest were 100% and 95.4%. The current National Committee for Clinical Laboratory Standards oxacillin breakpoint recommendation will categorise accurately the CoNS species encountered commonly.     

Comparison of Etest and agar dilution method for antimicrobial susceptibility testing of Flavobacterium isolates.

The Etest was evaluated as a possible alternative to the standard agar dilution method for susceptibility testing of nine antimicrobial agents against Flavobacterium species. In studies of 100 clinical isolates, the agreement between the MICs (+/-1 log2 dilution) obtained by the two methods was acceptable for cefotaxime, ceftazidime, amikacin, minocycline, ofloxacin, and ciprofloxacin (> 90%). Conversely, the agreement between the results obtained for piperacillin was limited (84%). The overall agreement was 92.5%.     

Comparison of the EUCAST-AFST broth dilution method with the CLSI reference broth dilution method (M38-A) for susceptibility testing of posaconazole and voriconazole against Aspergillus spp.

The susceptibilities of 40 clinical isolates of Aspergillus spp. (Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Aspergillus terreus) were determined for posaconazole and voriconazole by the CLSI M38-A and EUCAST-AFST broth dilution methods. Where a discrepancy was observed between the methods, the EUCAST method tended to give higher MIC values. Overall, the level of agreement was 92.5% and the intra-class correlation coefficient was > 0.90.     

Antimicrobial susceptibility testing of Helicobacter pylori to selected agents by agar dilution method in Shiraz-Iran

PURPOSE: To assess the pattern of antimicrobial susceptibility profile of Helicobacter pylori isolates from patients with gastritis, duodenal ulcer (DU) and gastroesophageal reflux disease (GERD) residing in Shiraz, Iran. METHODS: One hundred and six H. pylori isolates from patients with gastritis, DU and GERD undergoing endoscopy at our university hospitals and clinics were analysed for their antimicrobial susceptibility to metronidazole, clarithromycin, amoxicillin, co-amoxiclav, tetracycline, ciprofloxacin and furazolidone. The minimum inhibitory concentrations were determined by agar dilution method. RESULTS: Overall H. pylori resistance rate was 72.6% to metronidazole, 9.4% to clarithromycin and furazolidone, 20.8% to amoxicillin and 4.7% to tetracycline and ciprofloxacin. No resistance to co-amoxiclav was detected among H. pylori isolates. No significant differences between antimicrobial resistance and clinical outcome were detected. CONCLUSIONS: With regard to the increasing resistance of H. pylori isolates to various antibiotics, susceptibility testing of H. pylori isolates prior to the treatment of infection must be performed to achieve better eradication and to reduce the risk of selection of H. pylori resistant strains.     

Comparison of the E test and a reference agar dilution method for susceptibility testing of anaerobic bacteria

The susceptibility of 146 recent clinical isolates of gram-negative and gram-positive anaerobes was determined by the E test (AB Biodisk) on both Wilkins-Chalgren and PDM ASM II (AB Biodisk) agar. Results of the E test were compared with results obtained by the NCCLS agar dilution method using Wilkins-Chalgren agar. Incubation was for 20 hours and 44 hours in the E test and for 44 hours in the NCCLS method. In general, 44 hour results were more reliable; however, NCCLS readings were made only once after 44 hours. After two days of incubation, 91 % of E test results on Wilkins-Chalgren agar were within one dilution and 98 % within two dilutions of the corresponding NCCLS values; on PDM agar these values were 89 % and 98 %, respectively. Major and very major discrepancies combined were less than 1 %.     

Comparison of the Sceptor Pseudomonas Plus MIC Panel with agar dilution for susceptibility testing of Pseudomonas aeruginosa.

The antimicrobial susceptibilities of 100 clinical isolates of Pseudomonas aeruginosa to gentamicin, amikacin, tobramycin, ticarcillin, piperacillin, and ceftazidime were determined by using the Sceptor system (BBL Microbiology Systems, Cockeysville, Md.), and the results were compared with those obtained using the National Committee for Clinical Laboratory Standards reference agar dilution method. Excellent correlation was observed for the aminoglycosides, with greater than 95% agreement within 1 doubling dilution of the reference agar dilution MIC, while ticarcillin and piperacillin showed lower percent agreement values of 91 and 88%, respectively.     

In-vitro susceptibility testing by agar dilution method to determine the minimum inhibitory concentrations of amphotericin B, fluconazole and ketoconazole against ocular fungal isolates

Purpose: To standardize in-vitro antifungal susceptibility testing by agar dilution method to find out the minimum inhibitory concentration (MIC) of amphotericin B, fluconazole and ketoconazole on ocular fungal isolates. Methods: A total of 180 ocular fungal isolates (130 filamentous fungi and 50 yeasts) were included. The antifungal drugs such as amphotericin B (0.0625-8 μg/mL), fluconazole (0.2-819.6 μg/mL) and ketoconazole (0.025-6.4 μg/mL) were incorporated in doubling dilutions in the yeast nitrogen base medium. The MIC was determined as the lowest concentration of the antifungal drug preventing growth of macroscopically visible colonies on drug containing plates when there was visible growth on the drug - free control plates. Results: All 50 ocular isolates of yeast were susceptible to amphotericin B, while two (4%) and five (10%) strains were resistant to fluconazole and ketoconazole respectively. Of the 130 filamentous fungi tested, six (4.6%) were resistant to amphotericin B, 49 (37.7%) and 10 (7.6%) were resistant to fluconazole and ketoconazole respectively. Percentile 50 (MIC 50) and Percentile 90 (MIC 90) for all the three antifungal agents were calculated. Aspergillus niger, Aspergillus terreus and Candida krusei were found to be resistant to fluconazole and ketoconazole. Conclusion: This technique was found to be reliable, cost effective and easy to perform with consistent results.     

Comparison of Etest method with reference broth microdilution method for antimicrobial susceptibility testing of Yersinia pestis

The utility of Etest for antimicrobial susceptibility testing of Yersinia pestis was evaluated in comparison with broth microdilution and disk diffusion for eight agents. Four laboratories tested 26 diverse strains and found Etest to be reliable for testing antimicrobial agents used to treat Y. pestis, except for chloramphenicol and trimethoprim-sulfamethoxazole. Disk diffusion testing is not recommended.     

Comparison of the Sensititre YeastOne dilution method with the Clinical Laboratory Standards Institute (CLSI) M27-A3 microbroth dilution reference method for determining MIC of eight antifungal agents on 102 yeast strains

The Clinical Laboratory Standards Institute ([CLSI] formerly NCCLS) reference broth microdilution testing method (protocol M27-A3) was compared with a commercially available methods (Sensititre YeastOne^®) by testing two quality control strains and 102 isolates of Candida sp. and Cryptococcus sp. against fluconazole, itraconazole, ketoconazole, posaconazole, voriconazole, flucytosin, amphotericin B and caspofungin. Minimal inhibitory concentrations (MIC) endpoints were determined after 24 h of incubation for Sensititre YeastOne^® and after 24 and 48 h for CLSI microdilution method. Essential agreements between methods vary from 70.6 to 92.2%. Categorical agreements vary from 94.1% for 5FC to 72.6% for AMB. Sensititre YeastOne^® reading appears to be useful for avoiding very major errors and this confirms the interest of this method for evaluating new antifungals activity in vitro.     

Comparison of the Cobas-Bact five-hour susceptibility testing system with the NCCLS agar diffusion and dilution methods

The results of susceptibility tests performed by the Cobas-Bact system were compared with those of the NCCLS agar diffusion (Kirby-Bauer) and NCCLS agar dilution methods. A total of 998 clinical isolates were tested against 10 to 18 antimicrobial agents. Essential agreement (comprising full agreement and minor discrepancies) varied from 90.5 % to 99.2 % on comparison of Cobas-Bact with Kirby-Bauer results, depending on the bacterial group (mean for all 998 strains tested 95.7 %). These figures ranged from 91 % to 99.2 % (mean 96.3 %) for the Cobas-Bact/MIC comparison and from 95.2 % to 99.7 % (mean 98.7 %) for the Kirby-Bauer/MIC comparison. The best results were found forEnterobacteriaceae andStaphylococcus aureus, whereas for enterococci and coagulase-negative staphylococci there was a lower rate of essential agreement in all three comparisons. In the case ofPseudomonas aeruginosa there was a good rate of essential agreement but many minor discrepancies, resulting in a disappointing rate of full agreement of between 67.5 % and 78.9 % in the three comparisons. The Cobas-Bact system would appear to provide satisfactory susceptibility test results in most cases, however there are still some major problems in the system which should be resolved.     

Comparison of E test and agar dilution for antimicrobial susceptibility testing of Stenotrophomonas (Xanthomonas) maltophilia.

Currently recommended dilution test methods for the determination of antimicrobial susceptibility of Stenotrophomonas (Xanthomonas) maltophilia are labor-intensive and often impractical in many clinical laboratories. We compared the E test with the agar dilution method for susceptibility testing of 176 clinical isolates of S. maltophilia against 16 antimicrobial agents. The MICs obtained by E test correlated well with those determined by the agar dilution method, with an overall agreement of 94%. Very major and major errors occurred infrequently (0.6 to 2.9%) when testing beta-lactam agents, tobramycin, trimethoprim-sulfamethoxazole, and fluoroquinolones. The E test was found to be accurate and easy to perform. For most antimicrobial agents tested against S. maltophilia, the E test is an acceptable alternative susceptibility test method.     

Comparison of microdilution and agar dilution procedures for testing antibiotic susceptibility of Neisseria gonorrhoeae

Studies were run in parallel to compare the broth microdilution method and the chocolate agar dilution method for testing antibiotic susceptibility of Neisseria gonorrhoeae. Six clinically relevant drugs were tested against 23 clinical isolates of N. gonorrhoeae, including several penicillinase-producing, as well as multiply resistant, strains. Results showed that the MIC obtained by the two methods were not significantly different. The microdilution method appears to be a more sensitive system for discriminating penicillinase activity. The microdilution system is a more expedient method for screening new antibacterial agents and is more readily adaptable to new automated equipment.     

Quality control guidelines for testing cefotetan in the reference agar dilution procedure for susceptibility testing of anaerobic bacteria.

Reference values for quality control of in vitro susceptibility tests with cefotetan against anaerobic bacteria were determined in two independent multilaboratory studies with the approved National Committee for Clinical Laboratory Standards agar dilution method and three control strains (Bacteroides fragilis ATCC 25285, Bacteroides thetaiotaomicron ATCC 29741, and Clostridium perfringens ATCC 13124). The results of the two studies were in agreement. The recommended MIC control limits for B. fragilis ATCC 25285 and B. thetaiotaomicron ATCC 29741 are 4.0 to 16 micrograms/ml and 32 to 128 micrograms/ml, respectively. MICs for C. perfringens ATCC 13124 were too variable to be useful for controlling tests with cefotetan.     

Discrepancy in antimicrobial susceptibility test results obtained for oral streptococci with the Etest and agar dilution

A total of 270 viridans group streptococci (VS) isolated from healthy children, identified to the species level, were tested for their susceptibilities to penicillin, imipenem, erythromycin, and vancomycin. A total of 270 isolates and 1,080 organism-antibiotic combinations were evaluated. The overall susceptibility rates of all isolates obtained by the Etest (ET) versus agar dilution (AD) were 60.4% versus 61.8% for penicillin, 63.8% versus 63.9% for erythromycin, 90.6% versus 96% for vancomycin, and 99.1% versus 96.0% for imipenem, respectively. Major discrepancies occurred in the testing of the susceptibility of Streptococcus mutans to vancomycin, with 59.5% (ET) versus 100% (AD), followed by S. salivarius, with 84.1% versus 100%; S. oralis, with 82.1% versus 96.4%; and S. mitis, with 90% versus 100%, respectively. There were also differences in the rates of susceptibility of S. mutans, 66.5% (ET) versus 85.1% (AD), and S. intermedius, 82.9% versus 72.1%, respectively, to penicillin. General agreement between the results of ET and AD was obtained for 973 organism-antibiotic combinations out of 1,080 antibiotic combinations, i.e., 90.1%. Very major errors were found for 6.8% of isolates, and major errors were found for 3.2% of isolates; the minor errors were negligible. Agreement between the results of the two methods was 98.7% for penicillin, 94.6% for vancomycin, 96.9% for imipenem, and 99.9% for erythromycin. The highest rate of very major errors was for vancomycin, at 5.4%. The ET appears to be as efficient as AD for susceptibility testing of VS, except for vancomycin, where very major errors in the results were relatively high.     

Comparison of antimicrobial susceptibility testing of Campylobacter spp. by the agar dilution and the agar disk diffusion methods

The correlation and the level of agreement between the standardized agar dilution and the agar disk diffusion methods for antimicrobial susceptibility testing of Campylobacter were investigated. A high-level agreement between the two methods was evident for aminoglycosides and fluoroquinolones, while a low-level agreement was observed for other antibiotics.     

Inaccuracy of the disk diffusion method compared with the agar dilution method for susceptibility testing of Campylobacter spp

The agar dilution method has been standardized by the CLSI for the susceptibility testing of Campylobacter species, and according to these standards, the disk diffusion method should be used only in screening for macrolide and ciprofloxacin resistance. Nevertheless, the disk diffusion test is currently widely used, since it is easy to perform in clinical microbiology laboratories. In this study, the disk diffusion method was compared to the agar dilution method by analyzing the in vitro activities of seven antimicrobial agents against 174 Campylobacter strains collected in Finland between 2003 and 2008. Recommendations of the CLSI were followed using Mueller-Hinton agar plates with 5% of sheep blood. For each strain, the disk diffusion tests were performed two to four times. Of the 33 erythromycin-resistant strains (MIC, ≥16 μg/ml), 24 (73%) constantly showed a 6-mm erythromycin inhibition zone (i.e., no inhibition), while for seven strains the inhibition zone varied from 6 to 44 mm in repeated measurements. Among the 141 erythromycin-susceptible strains (MIC, <16 μg/ml), erythromycin inhibition zones varied between 6 and 61 mm. Of the 87 ciprofloxacin-resistant strains, 47 (54%) showed 6-mm inhibition zones, while 40 strains showed inhibition zones between 6 and 60 mm. Significant differences between the repetitions were observed in the disk diffusion for all antimicrobial agents and all strains except for the macrolide-resistant strains regarding the macrolides. For 17 (10%) strains, the variation in repeated measurements was substantial. These results show that the disk diffusion method may not be a reliable tool for the susceptibility testing of Campylobacter spp. Further studies are needed to assess whether the disk diffusion test could be improved or whether all susceptibilities of campylobacters should be tested using an MIC-based method.     

Evaluation of antibiotic susceptibility testing by agar dilution and the Micro Media system (Fox Panel).

The susceptibilities of 350 gram-positive cocci and 638 gram-negative bacilli to various antimicrobial agents were compared by using the Micro-Media system (MMS) (Fox Panel) (Micro-Media Systems, Inc., Potomac, Md.) and a standard agar dilution procedure. Major discrepancies occurred with enterococci, among which 48 of 53 isolates (91%) were found to be resistant to penicillin G by agar dilution and reported as susceptible by the MMS. Other large discrepancies occurred with Staphylococcus aureus and Acinetobacter calcoaceticus subsp. anitratus, among which more than 40% of the isolates were judged to be resistant to ampicillin by agar dilution and susceptible by the MMS. In terms of overall agreement in interpretation of MICs by the two systems, an agreement of greater than 84% was seen for both gram-positive and gram-negative organisms when ampicillin and cephalothin (68 and 78% agreement for gram-positive cocci, respectively) were excluded. These disagreements in MIC interpretations may result in part from the small number of organisms tested per well (4,000 CFU) in the MMS, as compared with 10,000 CFU per test in the agar dilution method.