Validation of the automated reading and incubation system with Sensititre plates for antimicrobial susceptibility testing

The present study compared the antimicrobial susceptibility testing (AST) results generated by the Automated Incubation and Reading System (ARIS) with custom Sensititre plates (TREK Diagnostic Systems, Cleveland, Ohio) and MicroScan PC10 GP and NUMIC10 GN plates interpreted with the WalkAway-96 system (Dade Behring, West Sacramento, Calif.) for gram-positive (GP) and gram-negative (GN) organisms as part of an in-house validation. A total of 326 isolates (3,689 antimicrobial agent-organism combinations) were evaluated. Sensititre plates were inoculated according to the instructions of the manufacturer with a suspension adjusted to a 0.5 McFarland standard, while the Prompt Inoculation System was used for the MicroScan plates. ARIS and the WalkAway system were used for automated reading of the Sensititre and MicroScan plates, respectively, at 18 to 24 h. The results were analyzed for essential (+/-1 twofold dilution) and categorical (sensitive, intermediate, or resistant) agreements. Plates that resulted in ARIS interpretations with major (falsely resistant) or very major (falsely susceptible) errors compared to the results obtained with the WalkAway system were read manually to corroborate instrument readings. Isolates for which very major or major errors were obtained and for which the results were not resolved by manual reading were retested in parallel. Isolates for which very major or major errors were obtained and for which the results were not resolved upon repeat testing were tested by the National Committee for Clinical Laboratory Standards M7-A5 frozen reference microdilution method. Essential agreement was 95.8% for 246 GN isolates. The following categorical error rates were obtained for the GN isolates: 1.3% minor errors, 0% major errors, and 0.4% very major errors. For 95 GP isolates, there was 93.5% essential agreement. Categorical error rates for GP isolates were 0.9% minor errors, 0.6% major errors, and 0.4% very major errors. ARIS-Sensititre is a diagnostic system feasible for use for automated AST in a clinical laboratory.     

Evaluation of the Merlin MICRONAUT system for rapid direct susceptibility testing of gram-positive cocci and gram-negative bacilli from positive blood cultures

Bloodstream infections are life-threatening conditions which require the timely initiation of appropriate antimicrobial therapy. We evaluated the automated Merlin MICRONAUT system for rapid direct microtiter broth antimicrobial susceptibility testing (AST) of gram-positive cocci and gram-negative bacilli from BACTEC 9240 bottles with positive blood cultures in comparison to the standard method for the Merlin MICRONAUT system. This prospective study was conducted under routine working conditions during a 9-month period. Altogether, 504 isolates from 409 patients and 11,819 organism-antibiotic combinations were evaluated for comparison of direct and standard AST methods. For gram-negative bacilli, direct and standard AST of 110 isolates was evaluated and MIC agreement was found for 98.1% of 2,637 organism-antibiotic combinations. Category (susceptible, intermediate susceptible, resistant [SIR]) agreement was found for 99.0%, with results for 0.04% of combinations showing very major errors, those for 0.2% showing major errors, and those for 0.8% showing minor errors. For gram-positive cocci, 373 isolates were evaluated and MIC agreement was found for 95.6% of 8,951 organism-antibiotic combinations. SIR agreement was found for 98.8%, with results for 0.3% of combinations showing very major errors, those for 0.4% showing major errors, and those for 0.5% showing minor errors. Although the number of tested isolates was limited (n = 33), direct AST of streptococci was performed for the first time, yielding promising results with SIR agreement for 98.6% of 363 organism-antibiotic combinations. In conclusion, direct AST of gram-negative bacilli and gram-positive cocci from positive blood cultures with the MICRONAUT system is a reliable technique that allows for the omission of repeat testing of subcultured isolates. Thereby, it reduces the time to results of blood culture testing and may have a positive impact on patient care.     

Use of positive blood cultures for direct identification and susceptibility testing with the vitek 2 system

In order to further decrease the time lapse between initial inoculation of blood culture media and the reporting of results of identification and antimicrobial susceptibility tests for microorganisms causing bacteremia, we performed a prospective study in which specially processed fluid from positive blood culture bottles from Bactec 9240 (Becton Dickinson, Cockeysville, Md.) containing aerobic media were directly inoculated into Vitek 2 system cards (bio-Mérieux, France). Organism identification and susceptibility results were compared with those obtained from cards inoculated with a standardized bacterial suspension obtained following subculture to agar; 100 consecutive positive monomicrobic blood cultures, consisting of 50 gram-negative rods and 50 gram-positive cocci, were included in the study. For gram-negative organisms, 31 of the 50 (62%) showed complete agreement with the standard method for species identification, while none of the 50 gram-positive cocci were correctly identified by the direct method. For gram-negative rods, there were 50% categorical agreements between the direct and standard methods for all drugs tested. The very major error rate was 2.4%, and the major error rate was 0.6%. The overall error rate for gram-negatives was 6.6%. Complete agreement in clinical categories of all antimicrobial agents evaluated was obtained for 19 of 50 (38%) gram-positive cocci evaluated; the overall error rate was 8.4%, with 2.8% minor errors, 2.4% major errors, and 3.2% very major errors. These findings suggest that the Vitek 2 cards inoculated directly from positive Bactec 9240 bottles do not provide acceptable bacterial identification or susceptibility testing in comparison with corresponding cards tested by a standard method.     

Rapid identification and antimicrobial susceptibility testing of gram-negative bacilli from blood cultures by the AutoMicrobic system.

A procedure was developed which allows direct identification and antimicrobial susceptibility testing of fermentative and nonfermentative gram-negative bacilli from positive blood cultures. A 10-ml sample was removed from turbid blood culture bottles, and the bacteria were washed and concentrated by centrifugation. The bacterial pellet was used to inoculate an Enterobacteriaceae Plus Identification Card and a Gram-Negative General Susceptibility Card of the AutoMicrobic system. Results with these cards were compared with results obtained with standard technique for 196 blood cultures seeded with recent clinical isolates. Identification of most cultures was available in 8 h, whereas the antimicrobial susceptibility results were available in an average of 4.7 h for all organisms. Direct identification was correct for 95% of the cultures, whereas the antimicrobial susceptibility data had an average agreement of 87% with 3.8% very major and 1.4% major errors. In using this procedure it was possible to provide accurate preliminary identification and results of antimicrobial susceptibility tests for gram-negative bacilli on the same day that a blood culture was determined to be positive.     

Evaluation of the BD Phoenix Automated Identification and Susceptibility Testing System in Clinical Microbiology Laboratory Practice

The Phoenix Automated Microbiology System (BD Biosciences, USA) is a new, fully automated system for the rapid identification and antimicrobial susceptibility testing of gram-positive and gram-negative bacteria. The objective of this study was to evaluate the quality of performance of the Phoenix system in the identification and antimicrobial susceptibility testing of 260 gram-negative (n=174) and gram-positive (n=86) isolates collected from Polish hospitals in recent years. Two Phoenix panel types for identification/antimicrobial susceptibility testing, NMIC/ID-5 for gram-negative rods and PMIC/ID-4 for gram-positive cocci, were used in the analysis according to the manufacturer's recommendations. The results produced by the system were compared with data obtained by reference or conventional microbiological methods. A high rate of agreement between the Phoenix and the conventional methods was observed for identification, ranging from 100% for gram-positive cocci to 96.0% for gram-negative nonfermenters and 92.5% for members of the family Enterobacteriaceae. Similarly, a high level of agreement characterized the antimicrobial susceptibility data obtained with the Phoenix and by the agar dilution method (2,361 test results). For staphylococci, enterococci and Enterobacteriaceae, the methods were 100% concordant in determining the category of susceptibility of isolates to the majority of the antimicrobial agents tested. A category agreement value of below 90% was found for the susceptibility of enterococci and gram-negative nonfermenters to ciprofloxacin (84.6% and 88.5%, respectively) and for susceptibility of Stenotrophomonas maltophilia to trimethoprim-sulfamethoxazole (80.0%).     

Evaluation of the Cobas-Bact system for direct and rapid identification and antimicrobial susceptibility testing of gram-negative rods from positive blood culture broths.

A direct antimicrobial susceptibility test and a direct identification of positive blood culture broths for gram-negative rods confirmed with Gram stain by using a new instrument, Cobas-Bact, were compared with the conventional Kirby-Bauer agar diffusion disk method and with the in-house set of identification or API 20E, respectively. The bacterial pellet of centrifuged positive blood culture broth was used to inoculate a Cobas-Bact susceptibility and identification rotor. Bacteria from 206 cases of monomicrobial septicemia due to members of the family Enterobacteriaceae were tested. In 198 episodes (96%), direct identification and antimicrobial susceptibility testing results were obtained for the same bacterial pathogen within 5 h of detection. Of 204 direct identifications obtained, 177 (86.6%) were "high-confidence" correct identifications (percentage of likelihood [P] greater than or equal to 80%) and 25 (12.5%) "low-confidence" correct identifications (P less than 80%), whereas only 2 misidentifications occurred (1 Escherichia coli and 1 Proteus mirabilis). Direct susceptibility testing was performed in 199 episodes (96%), providing 1,885 antibiotic-microorganism combinations. Full agreement reached 86.3%, and essential agreement reached 92.8%. Minor discrepancies were found in 120 (6.5%) of the tests, major discrepancies were found in 127 (6.8%) tests, and very major discrepancies were found in only 7 (0.4%) tests. Subsequent MIC determinations in cases of major or very major discrepancies reduced the number of major discrepancies involving cephalosporins from 60 to 16, whereas all those involving aminoglycosides remained. Overall, this direct and rapid Cobas-Bact identification and susceptibility testing procedure offered accurate information with 5 to 6 h after the laboratory detection of bacteremia and septicemia due to members of the Enterobacteriacease.     

Comparison of a highly automated 5-h susceptibility testing system, the Cobas-Bact, with two reference methods: Kirby-Bauer disk diffusion and broth microdilution.

The results of susceptibility tests performed with the Cobas-Bact system were compared with those of the Kirby-Bauer disk diffusion and the broth microdilution methods. The evaluation included tests with 24 antibiotics against 250 isolates of the family Enterobacteriaceae and 13 antibiotics against 100 gram-positive cocci. Complete agreements between the Cobas-Bact and Kirby-Bauer methods were 82.8 and 84.5% for gram-positive cocci and gram-negative bacilli, respectively. Agreements between the Cobas-Bact and broth microdilution methods were 76.7% for gram-positive cocci and 84.8% for gram-negative bacilli. Complete agreements between the Kirby-Bauer and broth microdilution methods were 87.0% for gram-positive cocci and 92.2% for gram-negative bacilli. Despite generally satisfactory results with most organism-antibiotic combinations tested, additional modifications of the Cobas-Bact system are required to reduce the number of major and very major discrepancies, as well as to permit testing of Pseudomonas spp. and other gram-negative nonfermentative bacilli.     

Evaluation of Alamar colorimetric broth microdilution susceptibility testing method for staphylococci and enterococci.

We compared the results of the Alamar broth microdilution susceptibility testing method with the results of the National Committee for Clinical Laboratory Standards reference broth microdilution method for 119 gram-positive organisms. The strains were tested for their susceptibilities to 20 antimicrobial agents. Only appropriate antimicrobial agents were evaluated for each species of bacteria. Absolute categorical agreement between the reference method and the test method was 91.5% for enterococci, 99.8% for oxacillin-susceptible staphylococci, and 97.4% for oxacillin-resistant staphylococci. Essential agreement (percent complete agreement plus percent minor errors) was > 99% for all organisms tested. The results for enterococci showed no very major errors, one major error with ofloxacin, and numerous minor errors with the quinolones. However, all except one of the minor errors were within +/- 1 log2 dilution of the reference result. For staphylococci, only 2 very major errors (one each with chloramphenicol and oxacillin), 1 major error (chloramphenicol), and 15 minor errors (multiple drugs) were observed. The Alamar colorimetric system was easy to use and the results were easy to read. It appears to be an acceptable method for antimicrobial susceptibility testing of staphylococci and enterococci.     

Direct inoculation on Phoenix panels for identification and antimicrobial susceptibility from positive BACTEC cultures: first study from India

Purpose: This was a prospective study planned in a super-specialty hospital in Delhi to reduce turnaround times of identification–susceptibility results of positive blood cultures. Materials and Methods: One hundred consecutive single morphology non-duplicate cultures were inoculated on Becton Dickinson Phoenix™ panels by growth recovered directly from liquid BACTEC™ media and after pure growth on solid media. Results: Complete concordance was observed in 72.4% of gram-negative and 45.8% of gram-positive isolates. For gram-negative isolates, categorical agreement (CA) was >83% and essential agreement (EA) was >96% among all antibiotics tested, very major errors (VME) were 0.13%, major errors (ME) 0.54%, and minor errors (MiE) were 3.01%. For gram-positive isolates, VME was 0.73%, 1.10% MiE and no ME. It was observed that average time from receipt of specimen to release of reports was 30:34 h and 32 h for gram-negative and gram-positive isolates if reports of “Direct” panels were to be released. Conclusions: By direct panel inoculation, a decrease of at least 18–20 h in turnaround time was observed compared with the standard method. This helps early change to effective antibiotic therapy and also reduces the expenditure incurred for a patient’s hospital stay by average Rs 20,000 ($443) per day.     

Clinical evaluation of automated antibiotic susceptibility testing with the MS-2 system.

The MS-2 (Abbott Laboratories) system for automated antimicrobial susceptibility testing was evaluated for both accuracy and general utility in our clinical laboratory. A total of 984 fresh clinical bacterial isolates (745 gram-negative, 239 gram-positive) were tested with the MS-2 system, and results were compared directly with those from a conventional agar disk diffusion method. Discrepancies between the two methods were categorized as very major, major, and minor. For gram-positive isolates, full accord (all discrepancies considered) was 91.6%, and essential accord (minor discrepancies not included) was 96.2%. With gram-negative isolates, full accord was found to be 93.9%, with essential accord of 97.9%. Aggrement as a function both of organism group and of antimicrobial agent was determined. Full accord of 90% or more was found for all major organism groups tested, with the exception of enterococci, where discrepant results between the two methods were observed. Mean test time for all isolates tested was 4.3 h. The MS-2 was found to be an accurate and highly automated instrument which required minimal technician time and was readily adaptable to work flow in our clinical laboratory.     

Antimicrobial susceptibility testing and profiling of Nocardia species and other aerobic actinomycetes from South Africa: comparative evaluation of broth microdilution versus the Etest

Nocardiosis is an underrecognized clinical entity in South Africa, for which interspecies epidemiological and clinical differences are poorly understood. The taxonomical state of flux and the lack of a simple antimicrobial susceptibility testing method are partly responsible. Definitive identification is molecularly based, which further complicates the study of this ubiquitous organism, as this methodology is beyond the scope of most routine diagnostic laboratories. The Etest methodology has been proposed as an alternative to the reference broth microdilution method, although there have been a limited number of comparative studies. We profiled 51 clinical isolates of aerobic actinomycetes, including 39 Nocardia species, using sequence-based (16S rRNA) identification. Broth microdilution and Etests were done concurrently on all isolates. The overall level of categorical and essential agreement for broth microdilution and Etest for the Nocardia isolates ranged from 67.5 to 100% and 46.2 to 81.6%, respectively. Very major errors were seen with amikacin, amoxicillin-clavulanate, ciprofloxacin, clarithromycin, and imipenem. For Nocardia species, uniform susceptibility to co-trimoxazole, amikacin, and linezolid was demonstrated, with a 48.8% susceptibility rate to imipenem. Nocardia farcinica (20.5%) and Nocardia cyriacigeorgica (15.4%) were the most commonly identified species among the 82% of isolates identified to species level using 16S rRNA sequences. Furthermore, drug susceptibility patterns demonstrated limited concordance with species identification. Our results suggest that, in a routine diagnostic setting, the Etest is not an acceptable alternative to the reference method of broth microdilution for antimicrobial susceptibility testing. Given the diversity and limited understanding of this group of organisms, further widespread evaluation of clinical isolates, from both clinical and diagnostic perspectives, is warranted.     

Rapid identification by the Micro-ID system of Enterobacteriaceae detected by urine screening.

Previous studies have demonstrated that organisms detected by urine screening can be processed for rapid identification and antimicrobial susceptibility testing directly from urine or urine screening broth. In the present study, an improved method for processing such specimens was evaluated. Organisms were harvested by centrifugation from positive urine screening broth, and inocula were prepared for rapid identification by the Micro-ID system and rapid susceptibility testing by the Autobac system. Nearly 2,500 urine specimens were analyzed by urine screening, and 206 specimens had significant growth of gram-negative, oxidase-negative bacilli. These organisms, prepared by the centrifugation procedure, were identified and tested for susceptibility to antimicrobial agents. For comparison, identifications by the Micro-ID system and antimicrobial susceptibility tests by the Autobac system were performed on the same organisms the next day with inocula prepared from colonies growing from standard urine cultures. The results demonstrated that 95% of the organisms were correctly identified by this procedure, and susceptibility testing by the rapid method gave results in 94% agreement with the standard method. These results demonstrate that organisms detected by urine screening can be accurately identified and tested for antimicrobial susceptibility after centrifugation from urine screening broth. This system provides a practical procedure or same-day reporting of urine culture results.     

Collaborative investigation of the accuracy and reproducibility of Sceptor Breakpoint susceptibility panels.

A combination Sceptor Breakpoint/ID panel (Johnston Laboratories, Inc., Towson, Md.), which determines interpretive susceptibility results (susceptible, moderately susceptible, and resistant) using two to three selected concentrations of antimicrobial agents, was tested in comparison with full-range Sceptor microdilution MIC panels. The inter- and intralaboratory interpretive reproducibilities for 24 control strains tested in three laboratories on three consecutive days were 97.0 and 95.7%, respectively. The equivalency of breakpoint results to category results obtained by the microdilution MIC procedure for 10,368 control organism-antimicrobial agent comparisons was 94.1%. The level of interpretive agreement between breakpoint and MIC category results using 101 fresh clinical isolates was 97.0% for 51 gram-negative and 50 gram-positive bacteria. Among the total 4,872 clinical organism-antimicrobial agent comparisons, major and very major discrepancies were seen in 0.2% of gram-negative bacteria and very major discrepancies were seen in 0.9% of gram-positive bacteria. All very major discrepancies with gram-positive organisms were associated with trailing endpoints using trimethoprim or sulfisoxazole and staphylococci. The breakpoint concept of testing selective antimicrobial agent concentrations was highly reproducible and accurate and allows for placement of more antimicrobial agents into a panel than is possible with full-dilution MIC testing.     

Automated reading of MIC microdilution trays containing fluorogenic enzyme substrates with the Sensititre Autoreader.

The Sensititre Autoreader is a microcomputer-driven instrument capable of automatically reading antimicrobial susceptibility microdilution trays. The instrument measures the fluorescence liberated by bacterial enzymatic activity on fluorogenic substrates as an indicator of growth in each well. A mathematical algorithm converts the fluorescent signals from an antimicrobial dilution series to an MIC endpoint. A three-center study evaluated the performance of the Autoreader in comparison with MIC determined visually in a duplicate set of control plates lacking fluorogenic substrate. Among 828 isolates of gram-negative bacilli tested against 17 antimicrobial agents, Autoreader 18-h MIC were within +/- 1 twofold dilution of control MIC values (agreement) in 95.3% of instances. In 3.5% of the instances, Autoreader values occurred +/- 2 half-step dilutions from control values (minor discrepancy), and in only 1.2% of instances did Autoreader values deviate from control values by greater than +/- 2 dilution steps (major discrepancy). Agreement, minor discrepancies, and major discrepancies were noted among 148 gram-positive cocci tested against 11 antimicrobial agents in 93.5, 4.8, and 1.7% of the instances, respectively. Over half of the major discrepancies noted with gram-negative bacilli occurred with Proteus mirabilis-beta-lactam combinations, a problem that was resolved when a lower initial inoculum was used. Inter-and intralaboratory reproducibility was excellent. Standard Sensititre susceptibility trays may be instrument read at 18 h reproducibly and accurately with only slight modification of conventional procedures to include fluorogenic enzyme substrates in the incubation broth.     

Evaluation of Etest and disk diffusion methods compared with broth microdilution antifungal susceptibility testing of clinical isolates of Candida spp. against posaconazole

We performed Etest, disk diffusion, and broth microdilution susceptibility testing of 2,171 clinical isolates of Candida spp. against posaconazole. By using provisional breakpoints for comparison purposes only, the categorical agreement between the agar-based methods and broth microdilution results ranged from 93 to 98%, with <1% very major errors. The essential agreement (within 2 well dilutions) between the Etest and broth microdilution methods was 94%. These agar-based methods hold promise as simple and reliable methods for determination of the posaconzole susceptibilities of Candida spp.     

Evaluation of the Wider system, a new computer-assisted image-processing device for bacterial identification and susceptibility testing

The Wider system is a newly developed computer-assisted image-processing device for both bacterial identification and antimicrobial susceptibility testing. It has been adapted to be able to read and interpret commercial MicroScan panels. Two hundred forty-four fresh consecutive clinical isolates (138 isolates of the family Enterobacteriaceae, 25 nonfermentative gram-negative rods [NFGNRs], and 81 gram-positive cocci) were tested. In addition, 100 enterobacterial strains with known beta-lactam resistance mechanisms (22 strains with chromosomal AmpC beta-lactamase, 8 strains with chromosomal class A beta-lactamase, 21 broad-spectrum and IRT beta-lactamase-producing strains, 41 extended-spectrum beta-lactamase-producing strains, and 8 permeability mutants) were tested. API galleries and National Committee for Clinical Laboratory Standards (NCCLS) microdilution methods were used as reference methods. The Wider system correctly identified 97.5% of the clinical isolates at the species level. Overall essential agreement (+/-1 log(2) dilution for 3,719 organism-antimicrobial drug combinations) was 95.6% (isolates of the family Enterobacteriaceae, 96.6%; NFGNRs, 88.0%; gram-positive cocci, 95.6%). The lowest essential agreement was observed with Enterobacteriaceae versus imipenem (84.0%), NFGNR versus piperacillin (88.0%) and cefepime (88.0%), and gram-positive isolates versus penicillin (80.4%). The category error rate (NCCLS criteria) was 4.2% (2.0% very major errors, 0.6% major errors, and 1. 5% minor errors). Essential agreement and interpretive error rates for eight beta-lactam antibiotics against isolates of the family Enterobacteriaceae with known beta-lactam resistance mechanisms were 94.8 and 5.4%, respectively. Interestingly, the very major error rate was only 0.8%. Minor errors (3.6%) were mainly observed with amoxicillin-clavulanate and cefepime against extended-spectrum beta-lactamase-producing isolates. The Wider system is a new reliable tool which applies the image-processing technology to the reading of commercial trays for both bacterial identification and susceptibility testing.     

Evaluation of the AutoMicrobic system for susceptibility testing of aminoglycosides and gram-negative bacilli.

The AutoMicrobic system (AMS; Vitek Systems, Inc., Hazelwood, Mo.) was compared with a reference broth microdilution MIC method to determine the accuracy and reproducibility of aminoglycoside susceptibility testing of gram-negative bacilli. Stock clinical isolates (n = 176) which demonstrated resistance to at least one aminoglycoside, extended-spectrum penicillin, or broad-spectrum cephalosporin (or a combination) were selected for this study. Isolates with moderate susceptibility to the aminoglycosides were also included. Of these isolates, 116 were either resistant or moderately susceptible to one or more of amikacin, gentamicin, netilmicin, and tobramycin. When AMS MIC results for 704 antimicrobial agent-organism combinations were compared with parallel microdilution MIC results, exact agreement (AMS MIC = reference MIC) rates were: amikacin, 71.6%; gentamicin, 71.6%; netilmicin, 83.0%; and tobramycin, 69.3%. Agreement rates within +/- 1 log2 dilution were: amikacin, 96.0%; gentamicin, 93.8%; netilmicin, 97.2%; and tobramycin, 96.0%. When National Committee for Clinical Laboratory Standards criteria were used to qualitatively evaluate performance, the overall agreement rates were: amikacin, 100.0%; gentamicin, 99.4%; netilmicin, 98.9%; and tobramycin, 99.4%. There were only four very major discrepancies, which represented 0.6% of the tests performed, and there were no major discrepancies. The percentages of minor discrepancies were: amikacin, 9.6%; gentamicin, 14.2%; netilmicin, 11.9%; and tobramycin, 10.8%. Of the overall average of 11.6% minor discrepancies, 9.7% occurred even though the AMS MIC was within +/- 1 log2 dilution of the reference MIC. The intralaboratory reproducibility ranged from 93.3 to 100% for the four drugs examined. With this challenge group of gram-negative bacilli, the AMS generated aminoglycoside MIC results that were comparable to those obtained by a reference broth microdilution method.     

Evaluation of the Cobasbact system for rapid antimicrobial susceptibility testing of positive blood culture broths

The automated Cobasbact system was adapted for direct antimicrobial susceptibility testing of positive blood culture broths and its performance compared with that of the conventional Kirby-Bauer agar disc diffusion method using 278 positive blood samples. Overall, 1746 antibiotic-organism combinations were tested. Full agreement was 86.9 % and essential agreement (i.e. including minor discrepancies) was 91.8 %. The system would seem to produce acceptable susceptibility results within five hours after detection of a positive blood culture broth.     

Comparison of the E Test to agar dilution, broth microdilution, and agar diffusion susceptibility testing techniques by using a special challenge set of bacteria.

The E Test (AB Biodisk, Solna, Sweden) is a new method for performing antimicrobial susceptibility tests. It consists of an impervious carrier (5- by 50-mm strip) with a predefined antimicrobic gradient which is placed on an inoculated agar plate and processed like a disk diffusion test. Results are generated directly as MICs from a continuous concentration gradient covering 15 twofold dilutions, and MICs are read where the edge of the inhibition zone intersects the strip. We compared the E Test with disk diffusion, broth microdilution, and agar dilution tests by using a challenge set of 195 gram-positive and gram-negative bacteria for 14 antimicrobial agents. Also, disk diffusion, broth microdilution, and agar dilution tests were compared with each other. All test method comparisons gave greater than 94% agreement for the category of susceptibility. The E Test category agreement with disk diffusion and broth microdilution was 95.1%, and with agar dilution it was 95.2%. The E Test results were as reliable as the results obtained by the standard antimicrobial susceptibility testing methods.     

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.