Normalized resistance interpretation as a tool for establishing epidemiological MIC susceptibility breakpoints

Normalized resistance interpretation (NRI) utilizes the fact that the wild-type population on the sensitive side is not affected by resistance development, and therefore a normalized reconstruction of the peak can be performed. The method was modified for MIC distributions by the introduction of helper variables, in-between values assigned the mean of the neighboring numbers of isolates. This method was used on Staphylo- coccus aureus and Escherichia coli MIC distributions for 27 antimicrobials each and obtained from the EUCAST (European Committee on Antimicrobial Susceptibility Testing) website (http://www.eucast.org/mic_distributions/). The number of isolates in each of the 54 distributions ranged from 40 to 124,472. NRI produced normalized distributions in all cases. Cutoff values were calculated for +2.0 and +2.5 standard deviations (SD) above the means and then rounded up to nearest regular MIC dilution step. EUCAST also show cutoff values, ECOFF values, which were used as the reference. The NRI generated +2.0 SD values showed the best agreement with 26 of 27 within ±1 dilution step and 17 exactly on the ECOFF values for Staphylococcus aureus, and 25 of 27 within ±1 dilution step and 14 right on the ECOFF values for Escherichia coli. NRI offers an objective method for the reconstruction of the wild-type population in an MIC distribution for a given bacterial species and an antimicrobial agent. This method offers a new tool in comparative susceptibility studies such as global surveillance of resistance, as well as in quality control in individual laboratories.     

Commercial broth microdilution panel validation and reproducibility trials for NVP PDF-713 (LBM 415), a novel inhibitor of bacterial peptide deformylase

NVP PDF-713 (LBM 415) is a peptide deformylase inhibitor being progressed into clinical trials. Dry-form broth microdilution panels of NVP PDF-713 were compared to reference MIC panels of 552 recent clinical isolates. Most (99.2%) dry-form MIC results were within +/- 1 log(2) dilution of the reference panel MICs. Of the bacteria tested, Streptococcus pneumoniae and Haemophilus influenzae showed a bias towards higher and lower MICs, respectively. Same-day and between-day reproducibility tests showed that 98.9% and 96.7% of MIC values, respectively, were within +/- 1 log(2) dilution step, thereby demonstrating a high degree of reliability of the dry-form MIC product for clinical studies.     

Evaluation of mupirocin E-test for determination of isolate susceptibility: comparison with standard agar dilution techniques.

Mupirocin E-test strips have been evaluated for their ease of use and accuracy in determining the susceptibilities of 171 strains of Staphylococcus spp., Streptococcus spp., Haemophilus influenzae, and Moraxella catarrhalis. The susceptibility of each strain was determined on two occasions, using parallel E-test and agar dilution methodologies each time. To ensure similar precisions for statistical analyses, E-test MICs were rounded up to a standard twofold agar dilution scale. Clear, elliptical zones were obtained against Staphylococcus spp. M. catarrhalis also gave clear zones, but the scale intercept was often difficult to interpret because of the irregular shape of the inhibition zone. Poor growth sometimes resulted in less-distinct zones of inhibition against Streptococcus spp. and H. influenzae. Excellent correlation was observed between the the E-test and agar dilution against Staphylococcus spp. and H. influenzae, with > 95% of the E-test values falling within one log2 dilution of the corresponding agar MIC. The correlation was lower for Streptococcus spp. and M. catarrhalis, with 86 and 83%, respectively, of E-test results falling within one log2 dilution of the agar MIC. When E-test MICs did not agree exactly with the corresponding agar MIC against Staphylococcus spp. or Streptococcus spp., there was a tendency for the E-test to give a lower MIC. This bias has little effect upon individual MICs in staphylococci or in the generation of susceptibility interpretation errors ( < 1.5% overall), but it could reduce population geometric mean MICs by factors of 0.78 to 0.83.(ABSTRACT TRUNCATED AT 250 WORDS)     

Evaluation of in vitro spectra of activity of azithromycin, clarithromycin, and erythromycin tested against strains of Neisseria gonorrhoeae by reference agar dilution, disk diffusion, and Etest methods.

The macrolide-azilide susceptibility testing (agar dilution, disk diffusion, Etest) criteria for 105 Neisseria gonorrhoeae strains were evaluated. In addition, the potencies of azithromycin, clarithromycin, and erythromycin were studied. The most active macrolide-azilide agent was azithromycin (MIC at which 90% of the isolates are inhibited [MIC90], 0.5 microgram/ml) compared with clarithromycin (MIC90, 1.5 to 2 micrograms/ml) and erythromycin (MIC90, 2 to 4 micrograms/ml). The Etest (AB Biodisk, Solna, Sweden) was observed to produce MIC results very similar to those of the reference agar dilution test (GC agar base), with 100% of the results within 1 log2 dilution step of the reference MICs. The disk diffusion test zone diameters for all three drugs correlated at an acceptable level (r = -0.81 to -0.92) with the reference agar dilution MICs. Interpretive criteria for susceptibility were proposed for azithromycin at a MIC of < or = 2 micrograms/ml and a disk diffusion test zone of > or = 25 mm. No category for resistance was proposed because of the paucity of strains for which MICs were > 2 micrograms/ml. These tentative criteria should be further validated by correlations with clinical trial data for gonococcal strains (as they emerge) that have azithromycin MICs above the proposed susceptible category range.     

Preliminary evaluation of a semisolid agar antifungal susceptibility test for yeasts and molds

This report presents a semisolid agar antifungal susceptibility (SAAS) method for the rapid susceptibility screening of yeasts and molds. The reproducibility and accuracy of the SAAS method were assessed by comparing the MICs of amphotericin B and fluconazole obtained for 10 candidate quality control (QC) American Type Culture Collection yeast strains in >/=15 replicates with those found by six independent laboratories using the National Committee for Clinical Laboratory Standards (NCCLS) M27-P broth macrodilution method (M. A. Pfaller et al., J. Clin. Microbiol. 33:1104-1107, 1995). Overall, 96% of MICs for both drugs fell within 1 log(2) dilution of the modal MIC for each strain. The MICs for amphotericin B showed 99% agreement with the NCCLS proposed QC ranges within 1 log(2) dilution. Likewise, the MICs for fluconazole at >/=75% growth reduction showed 99% agreement for seven strains. Three strains, Candida albicans ATCC 24333 and ATCC 76615 and Candida tropicalis ATCC 750, showed a less sharp fluconazole endpoint at >/=75% growth reduction, but at >50% growth reduction, the agreement was 98% within 1 log(2) dilution of the proposed range. The MIC agreement within the proposed range for the suggested QC strains Candida parapsilosis ATCC 22019 and Candida krusei ATCC 6258 was 100% for fluconazole and 100% within 1 log(2) dilution of the proposed range for amphotericin B. The SAAS method demonstrated the susceptibility or resistance of 25 clinical isolates of filamentous fungi such as Aspergillus fumigatus to amphotericin B, itraconazole, and fluconazole, usually within 48 h. Although the results are preliminary, this SAAS method is promising as a rapid and cost-effective screen and is worthy of concerted investigation.     

Commercial broth microdilution panel validation and reproducibility trials for garenoxacin (BMS-284756), a novel desfluoroquinolone

Results from garenoxacin dry-form broth microdilution MIC panels prepared commercially (Sensititre, TREK Diagnostics) were compared to reference frozen-form MICs to ensure the validity of the longer-shelf-life product. A total of 1078 organisms from seven major organism groups were used in this trial. All commercial MIC results were within +/- one log(2) dilution of reference garenoxacin values, and reproducibility trials produced identical MIC results for 90.5 to 92.1% of garenoxacin MIC comparisons. Control quinolones (ciprofloxacin and gatifloxacin) also performed at a similarly high level of accuracy.     

Comparison of anaerobic susceptibility results obtained by two methods of inoculum preparation

We evaluated the use of inocula prepared directly from blood agar plates in agar dilution susceptibility tests of anaerobic bacteria and compared the results with susceptibility results obtained from the National Committee for Clinical Laboratory Standards proposed thioglycolate broth cultures. The objectives were to evaluate the reproducibility of each of the two methods of inoculum preparation and to compare the MICs obtained by each method. The reproducibility studies were conducted on 14 stock strains. The mode MICs obtained by the direct agar method were identical to those obtained by the reference broth method 74% of the time and within +/- 1 log2 dilution 100% of the time. The degree of reproducibility of each of the two methods was identical (93% +/- 1 log2 dilution). MIC results obtained by the direct agar method agreed with the MICs obtained by the reference broth culture method in 92.9% of 1,125 MIC data pair determinations performed on stock cultures. The reproducibility of the direct agar method within +/- 1 log2 dilution step for 115 fresh clinical isolates was 93%, including 93.4% of the results with the Bacteroides fragilis group. Only two very major discrepancies (false-susceptible by the agar method) were identified among the 708 MIC data pairs on these clinical isolates. Preparation of inocula directly from growth on agar plates provides a rapid and reproducible method for agar dilution susceptibility testing of anaerobes.     

Differences in vancomycin MIC among MRSA isolates by agar dilution and E test method

In this study, the correlation between vancomycin minimum inhibitory concentration (MIC) obtained by the E test technique and the Clinical And Laboratory Standards Institute (CLSI) agar dilution method was evaluated. A total of 53 Methicillin Resistant Staphylococcus aureus (MRSA) strains were tested by both the methods in the present study. MICs of vancomycin obtained by the E test method were consistently higher (+0.5 to 2 log2 dilutions) than those obtained by the agar dilution method. Out of 53 MRSA isolates, 49 isolates showed higher MIC results by E test than by agar dilution method. Three isolates showed same MIC result by both methods. Since many studies have demonstrated increased clinical failure with MRSA isolates for which vancomycin MICs are increased (>1 µg/ml) but still within the susceptibility range (≤ 2 µg/ml), our findings suggest the requirement to re-look into the breakpoints for vancomycin for determining sensitivity of MRSA isolates. Guidelines should also specify the method to be used for determining the MIC.     

Evaluation of the MICUR system for quantitative antimicrobial susceptibility testing: a multiphasic comparison with reference methods.

Four laboratories participated in a three-phase study to evaluate the MICUR antimicrobial broth microdilution system (Boehringer Mannheim Diagnostics, Inc., Houston, Tex.). The dried-antimicrobial agent MICUR system was compared with a reference broth microdilution method (National Committee for Clinical Laboratory Standards) by using 304 recently isolated clinical strains and two collections of stock or challenge organisms. Of 7,092 minimum inhibitory concentration (MIC) datum pairs derived from the clinical isolates, 96.6% were within an acceptable (+/- 1 log2 dilution) range. MICUR MICs agreed with the reference broth microdilution method MICs in 95.3% of 6,840 MIC pair determinations performed on stock or challenge cultures. The MICUR intralaboratory reproducibility within +/- 1 log2 dilution step for the clinical isolates was 98.4%. The MICUR intralaboratory and interlaboratory reproducibilities for 26 stock cultures were 98.4 and 95.1%, respectively. For 180 challenge cultures (4,199 MIC pairs) which were included in the MICUR testing to provide a wide variety of antimicrobial susceptibility and resistance patterns, the results for 92.5% were in close agreement with the reference broth microdilution results. No specific resistance mechanism went unrecognized by this new commercial system. The MICUR system gives comparable MIC results when evaluated against the reference broth microdilution method, and it would be acceptable for use in clinical microbiology laboratories.     

Detection of penicillin and extended-spectrum cephalosporin resistance among Streptococcus pneumoniae clinical isolates by use of the E test.

Increasing penicillin resistance and the initial recognition of resistance to extended-spectrum cephalosporins among Streptococcus pneumoniae isolates have placed greater emphasis on accurate methods for susceptibility testing of clinical isolates. This study has evaluated the use of the E test (AB Biodisk NA, Piscataway, N.J.) for the detection of penicillin and cefotaxime resistance among 147 pneumococcal clinical isolates in three geographically separate laboratories. These included 42 penicillin-resistant (MIC, > or = 2 micrograms/ml) and 14 cefotaxime-resistant (defined here as an MIC of > or = 2 micrograms/ml) isolates. E test strips were applied to the surface of Mueller-Hinton sheep blood agar plates and incubated at 35 degrees C in 5% CO2 for 20 to 24 h. E test MICs were compared with MICs determined with lysed horse blood-supplemented Mueller-Hinton broth in a microdilution format as recommended by the National Committee for Clinical Laboratory Standards. Penicillin MICs agreed within one log2 dilution for 136 of 147 (92.5%) isolates, and cefotaxime MICs agreed within one log2 dilution for 142 of 147 (96.6%) isolates. No very major or major interpretive errors occurred with either penicillin or cefotaxime E test MIC results. There were 9.5 and 5.4% minor interpretive category errors with penicillin and cefotaxime E test MICs, respectively. These data indicate that the E test represents a convenient and reliable method for the detection of penicillin or cephalosporin resistance in pneumococci.     

MIC determination of fusidic acid and of ciprofloxacin using multidisk diffusion tests

Objective   To investigate the possibility of estimating the MICs of fusidic acid and ciprofloxacin for bacterial isolates using series of antibiotic disk concentrations in diffusion tests, so-called M-tests.
Methods   Thirty Staphylococcus aureus and S. epidermidis strains were tested for fusidic acid susceptibility. Sixty-one clinical isolates of eight bacterial species were tested for ciprofloxacin susceptibility. Disk diffusion was standardized according to the Swedish reference group for antibiotics (SRGA). For fusidic acid, a series of disks (1.5, 5.0, 15, 50 and 150 µg) was used. Ciprofloxacin was applied in four different diffusion sources (1, 3, 10 and 30 µg) on a single strip, the M-strip, and used. True MIC values were determined using the standardized agar dilution method according to the SRGA. Single-strain regression analysis (SRA) was employed to calculate critical concentration equivalents ( Q _zero).
Results   Fusidic acid and ciprofloxacin critical concentrations were determined for the bacterial isolates. The mean conversion factors for Q _zero to yield the true MIC were 2.06 (range 0.34–8.9) for fusidic acid and 2.05 (range 0.37–8.1) for ciprofloxacin. There was a correlation between true MIC values (all MICs expressed as 2 log + 9) and the calculated MIC values ( Q _zero× conversion factor) for both fusidic acid ( R  = 0.9822) and ciprofloxacin ( R  = 0.9696).
Conclusions   MIC values of clinical isolates can be estimated using SRA calculations on zone measurements in disk tests with several concentrations of the antibiotic in diffusion sources.     

Multicentre testing of the EUCAST broth microdilution reference method for MIC determination on Mycobacterium tuberculosis

ObjectivesThe first objective of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) subcommittee for antimycobacterial susceptibility testing (AMST), launched in 2016, was to set a reference method for determining the MICs of antituberculous agents, since many protocols are used worldwide and a consensus one is needed for the determination of microbiological breakpoints.MethodsDuring 2017 and 2018, MIC determination protocols were evaluated prospectively in a multicentre study within the four AMST laboratories. MIC results were obtained for isoniazid, levofloxacin and amikacin on the reference strain Mycobacterium tuberculosis H37Rv ATCC 27294. Broth microdilution (BMD) in Middlebrook 7H9 and solid medium dilution (SMD) in Middlebrook 7H10 were performed using two inoculum concentrations. MICs were interpreted with regard to visual and 99% inhibition after 7, 14 or 21 days of incubation for BMD and 21 days for SMD.ResultsFollowing the EUCAST reference protocol, intra- and inter-assay agreements were within ±1 MIC dilution for >95% of the observations for the three drugs in both methods. MIC values, presented as MIC mode (range) for BMD and SMD respectively, were: 0.03 (0.015–0.06) mg/L and 0.12 (0.06–0.25) mg/L for isoniazid, 0.25 mg/L (0.25–0.5) and 0.5 mg/L (0.12–0.5) for levofloxacin, and 0.5 mg/L (0.5–1.0) and 0.5 mg/L (0.5–1.0) for amikacin.ConclusionsBoth SMD and BMD were reproducible and eligible as a reference method for MIC determination of the Mycobacterium tuberculosis complex (MTBC). BMD was finally selected as the EUCAST reference method. From now on it will be used to set epidemiological cut-off values and clinical breakpoints of new and old antituberculous agents.     

Antimicrobial susceptibility of multidrug-resistant Gram negative bacteria to fosfomycin

We evaluated the antimicrobial activity of fosfomycin against a randomly selected sample of 30 Klebsiella pneumoniae, 30 Pseudomonas aeruginosa, and 30 Acinetobacter baumannii multidrug-resistant, clinical isolates from patients in a general tertiary care hospital in Athens, Greece. Standard laboratory methods were used for susceptibility testing to commonly used antibiotics and the detection of extended-spectrum-beta-lactamase (ESBL) and metallo-beta-lactamase (MBL) production. The minimum inhibitory concentration (MIC) of fosfomycin for each isolate was determined by the agar dilution method. All K. pneumoniae isolates were both ESBL and MBL producers; all P. aeruginosa isolates were ESBL producers. The K. pneumoniae strains had fosfomycin MICs distributed across a range of 8-64 microg/ml; MIC(50) was 16 microg/ml and MIC(90) 32 microg/ml. The fosfomycin MICs of the P. aeruginosa strains had a distribution across a range of 4 to over 512 microg/ml; MIC(50) was 32 microg/ml and MIC(90) 128 microg/ml. The fosfomycin MICs of the A. baumannii strains had a distribution across a range of 64 to over 512 microg/ml; MIC(50) was 256 microg/ml and MIC(90) more than 512 microg/ml. Although standardized fosfomycin MIC interpretative breakpoints for the species studied are lacking, the findings of our study support the idea that fosfomycin may be further investigated as one among a decreasing list of therapeutic options for the treatment of infections due to multidrug-resistant strains of, primarily, K. pneumoniae and, secondly, P. aeruginosa.     

Comparison of broth microdilution,E Test,and agar dilution methods for antibiotic susceptibility testing of Campylobacter jejuni and Campylobacter coli

A standardized broth microdilution method was compared to the E test and an agar dilution method for the antimicrobial susceptibility testing of Campylobacter jejuni and C. coli isolates. A group of 47 human clinical isolates, 37 isolates from retail poultry, and 29 isolates from living turkeys (total, 113 isolates) was included in the study. These encompassed 92 C. jejuni and 21 C. coli strains. The MICs of six antimicrobial agents were determined by the broth microdilution and E test methods, and the strains of human origin were additionally tested by the agar dilution method. In general, broth microdilution MICs agreed within 1 log(2) MIC increment with 90.0% of E test results and 78.7% of agar dilution test results. The agar dilution method gave much lower gentamicin MICs than the broth microdilution method, but the data were significantly (P < 0.01) correlated and there was 100% agreement in the sensitivities and specificities in the comparison of the tests. The broth microdilution method had the highest sensitivity for analysis of the susceptibilities of Campylobacter to nalidixic acid and trimethoprim-sulfamethoxazole. The MICs of ciprofloxacin and erythromycin complied numerically by all three methods. The classification of the results and the correlation of the data demonstrated a high degree of agreement. All methods were equally suitable for the testing of the sensitivity of Campylobacter to tetracycline. Thus, the broth microdilution method appears to be an easy and reliable method for determination of the MICs of antibiotics for C. jejuni and C. coli, and it may offer an interesting alternative to MIC determination by the agar dilution technique or the E test.     

Proposed MIC quality control guidelines for National Committee for Clinical Laboratory Standards susceptibility tests using seven veterinary antimicrobial agents: ceftiofur, enrofloxacin, florfenicol, penicillin G-novobiocin, pirlimycin, premafloxacin, and spectinomycin.

The present multicenter study proposes broth microdilution quality control (QC) ranges for the antimicrobial agents ceftiofur, enrofloxacin, florfenicol, penicillin G-novobiocin, pirlimycin, premafloxacin, and spectinomycin, which are used in veterinary practice. Six separate laboratories tested replicates of National Committee for Clinical Laboratory Standards (NCCLS)-recommended QC organisms (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 29213, and Enterococcus faecalis ATCC 29212) on medium lots both common and unique to all laboratories. The proposed ranges were within 3 or 4 log2 dilution steps of the modal MICs for all organism-antimicrobial agent pairs, depending on their MIC distributions. With > or = 94.7% of all MIC results being within the proposed QC ranges, all combinations tested comply with NCCLS guidelines and all have been accepted by the NCCLS subcommittee developing susceptibility testing procedures for veterinary laboratories.     

Accuracy of the E test for determining antimicrobial susceptibilities of staphylococci, enterococci, Campylobacter jejuni, and gram-negative bacteria resistant to antimicrobial agents.

We compared the results of the E test MIC method with the results of agar dilution susceptibility testing for 18 antimicrobial agents against 324 strains of gram-positive and gram-negative bacteria, including 99 strains of staphylococci, 101 strains of antimicrobial-resistant gram-negative bacteria, 40 strains of enterococci, and 84 isolates of Campylobacter jejuni. Overall agreement of MICs (+/- 1 log2 dilution) was 97.3% for staphylococci, 94.6% for gram-negative bacilli, and 100.0% for enterococci. The MIC results for C. jejuni showed an overall agreement of only 82.9%. This was due primarily to a number of offscale values that limited the number of comparisons with clindamycin, trimethoprim-sulfamethoxazole, and tetracycline. Interpretative criteria for the results of the two test methods, however, were similar. Overall, the E test produced MIC results comparable to those of agar dilution when multiresistant organisms were tested. However, it was necessary to add 2% NaCl to the agar when testing oxacillin against staphylococci for both the E test and agar dilution to obtain results comparable to those of the broth microdilution method.     

Analysis of antimicrobial susceptibility results using microcomputers.

A computer program has been developed on an IBM-PC-compatible microcomputer to analyze antimicrobial susceptibility results of large numbers of organisms tested against a variety of antimicrobial agents. For each species, the output includes the numbers, percentages, cumulative numbers, and cumulative percentages of organisms inhibited at different concentrations of each drug, the MICs for 50 and 90% of organisms tested, MIC ranges, and geometric mean MICs. The distributions of MICs are also plotted as histograms. The program was written in dBASE III command language, but a compiled version allows rapid program execution independently of the dBASE III package.     

In vitro susceptibility of Listeria monocytogenes isolated from human blood and cerebrospinal fluid. A material from the years 1958-1985

The in vitro susceptibility of 156 strains of Listeria monocytogenes isolated since 1958 from human cerebrospinal fluid or blood to twelve antibiotics was determined by an agar dilution technique. Erythromycin (0.05), trimethoprim (0.2), netilmicin (0.2), and penicillin (0.2) were the most active drugs on weight basis (MIC90 0.05-0.2 micrograms/ml). Ampicillin and imipenem had MICs for 90% of the strains of 0.4 micrograms/ml. Ceftazidime was inactive (MIC90 greater than 100 micrograms/ml). Comparison of susceptibility pattern between strains isolated in different years showed that the antimicrobial susceptibility of L. monocytogenes has not changed during the last 25 years. The minimal bactericidal concentration (MBC) of penicillin was determined by a macro tube dilution method in ten recent isolates. Penicillin was bactericidal for all the strains with a MBC of 0.4-3.1 micrograms/ml, i.e. one to three two-fold dilutions above the MIC of 0.2-0.8 micrograms/ml, which means that no tolerant strains were found.     

Multicenter study of ofloxacin activity on bacteria isolated from a hospital environment

Minimal inhibitory concentrations (MICs) of ofloxacin were evaluated by agar dilution for 1508 bacterial strains isolated in five hospitals. For Enterobacteriaceae sensitive to nalidixic acid, MICs ranged from 0.008 to 1 microgram/ml (mode MIC: 0.12); the different species of Enterobacteriaceae exhibited similar mode MICs (0.12) with the exception of E. coli (0.06-0.12), P. mirabilis (0.5) and Providencia (0.25). Among strains intermediate and resistant to nalidixic acid, most of which were Serratia, Providencia and Citrobacter, 41% had a MIC within the susceptibility range, while the others had a MIC of 2 to 8 micrograms/ml, or even 64 micrograms/ml in a few instances. Ofloxacin also exhibited satisfactory activity against P. aeruginosa, with MICs ranging from 0.25 to 16 micrograms/ml (mode MIC: 2) for 87% of strains, and A. calcoaceticus, with MICs from 0.25 to 2 micrograms/ml (mode MIC: 1). Haemophilus sp. (MIC: 0.008 to 0.06 microgram/ml; mode MIC: 0.03), Gonococci (mode MIC: 0.008), and Meningococci (mode MIC: 0.016) were very sensitive to ofloxacin. The spectrum of ofloxacin included Gram positive cocci: MICs of Staphylococci were 0.06 to 2 micrograms/ml (mode MIC: 0.5); Enterococci, other Streptococci and Pneumococci were less sensitive, with MICs of 2 to 4 micrograms/ml for the majority of strains. As for anaerobic bacteria, ofloxacin proved more active against Clostridium (0.5 to 2 micrograms/ml) than Bacteroides (0.5 to 16 micrograms/ml).     

Evaluation of the E test for quantitative antimicrobial susceptibility testing of Helicobacter pylori.

The Progressive Diagnostics Manufacturers epsilometer test (E test; AB Biodisk, Solna, Sweden), a quantitative variant of the disk diffusion technique, was evaluated comparatively to an agar dilution method for the antimicrobial susceptibility testing of Helicobacter pylori. A collection of 79 H. pylori clinical strains, including isolates with known resistance to various antimicrobial agents, was tested against 12 different antimicrobial agents. All strains were tested on Columbia agar supplemented with 10% horse blood. Plates were incubated at 37 degrees C in microaerobic atmosphere (5% O2, 10% CO2), and readings were done after 3 days of incubation. In general, E test MICs were easy to interpret and the correlation between MICs by the agar dilution method and the E test was good, with 86 and 99.5% of results being within, respectively, 1 and 2 log2 dilution steps in a total of 936 tests. All strains of H. pylori with documented resistance to the tested agents were detected by the E test. Thus, the E test appears to be an easy and reliable method for determination of MICs of antibiotics for H. pylori, and it may offer an interesting alternative to MIC determination by the agar dilution technique.