Performance of enterobacteriaceae identification systems. An analysis of College of American Pathologists Survey data.

Commercial systems designed and marketed for evaluation of Enterobacteriaceae are used in many clinical microbiology laboratories. Evaluations of these systems have been reported from several large laboratories. Identifications with one or more of these systems were compared with those obtained by conventional methods with many strains of organisms. The Microbiology Resource Committee of the College of American Pathologists designed two surveys to evaluate the performances of commercial systems with identical strains of organisms submitted to a large number of participants. The surveys used were the Special Bacteriology Survey D-D and the Comprehensive Microbiology Survey B-D submitted during 1975--1976. Single strains of Klebsiella pneumoniae and Serratia marcescens in pure culture were submitted in the Comprehensive Survey and the same strains of these organisms plus one strain each of Pseudomonas stuartii and Citrobacter freundii in the Special Survey. The data obtained from participants permitted comparisons to be made between the API-20E, Enterotube, and r/b systems. These comparisons included individual biochemical test results shared by at least two systems as well as the ability of each to identify the unknown organisms to genus alone and to both genus and species. The results are presented and subjected to statistical analysis.

Direct comparison of two mechanized systems for identification of gram-negative bacilli. Autobac ID system versus the auto microbic system (with EBC plus).

The Auto Microbic System (AMS) is an almost completely automated system, capable of identifying Enterobacteriaceae after 8 hours, and some glucose nonfermenters after 13 hours of incubation. The Autobac ID system is a mechanized, computer-assisted system capable of identifying Enterobacteriaceae and many nonfermentative gram-negative bacilli within 3-6 hours. The present report combines the results of two independent studies, both of which evaluated the AMS and the Autobac ID system compared with standard reference tests. Among the 1,510 isolates that were tested, both systems reported equivocal identifications (low confidence values) with 5-6% of the strains. AMS produced fewer erroneous identifications (3.8% vs. 4.9%) but more equivocal test results (5.7% vs. 4.9%). Reproducibility of the two systems was compared by triplicate testing of 88 selected strains in both laboratories. AMS was somewhat more reproducible than the Autobac ID system. The AMS was capable of identifying more species with greater accuracy and reproducibility, but the Autobac ID system was more rapid. Both systems demonstrated excellent accuracy and reproducibility and both could be used efficiently in the clinical laboratory.

Reassessment of the "class" concept of disk susceptibility testing. Cephalothin disks versus minimal inhibitory concentrations with eleven cephalosporins.

Reassessment of the "class" concept of disk susceptibility testing. Cephalothin disks versus minimal inhibitory concentrations with eleven cephalosporins. Am J Clin Pathol 70: 909--913, 1978. Studies were carried out to determine whether susceptibility or resistance to 11 cephalosporins could be predicted reliably from the results of tests with a single cephalothin disk. The cephalosporins were tested with a microdilution technic and with a standardized disk test. Strains susceptible to a cephalothin disk were predictably susceptible to all other cephalosporins. However, 2--12% of the strains were resistant to cephalothin disks but were susceptible to the more active parenteral drugs cefoxitin, cephamandole, cefuroxime, and BL-S786. Because of differences in antimicrobial activities, the cephalosporins could be divided into three subgroups for purposes of susceptibility testing: one subgroup includes the majority of cephalosporins and may be represented by tests with cephalothin, the second subgroup inclues three active parenteral drugs (cephamandole, cefuroxime, and BL-S786) and may be represented by tests with cefuroxime, and the third subgroup consists of cefoxitin, a cephamycin with a unique broad spectrum of activity. Until the drugs in the second and third subgroups are released for general therapeutic use, the practice of testing only one cephalosporin disk appears to be a reasonably reliable procedure.

In vitro activity of carumonam (RO 17-2301), BMY-28142, aztreonam, and ceftazidime against 7,620 consecutive clinical bacterial isolates.

For 45-60 days four geographically separate clinical laboratories tested routine clinical bacterial isolates for susceptibility to carumonam, aztreonam, BMY-28142, and ceftazidime by the broth microdilution method. All four drugs were highly active against Enterobacteriaceae, inhibiting greater than 96% of the 4887 strains tested at less than or equal to 8.0 microgram/ml. The minimal inhibitory concentration at which 50% of the isolates were inhibited for each drug was less than or equal to 0.125 micrograms/ml. Ceftazidime was the most active against nonenteric gram-negative bacilli (86% inhibited at less than or equal to 8.0 micrograms/ml), followed by BMY 28142 (82%), carumonam (75%), and aztreonam (68%). The two monobactams exhibited no activity against gram-positive cocci at the concentrations tested, whereas BMY-28142 had excellent activity against nonenterococcal streptococci and good activity against staphylococci.

In vitro evaluation of cefixime (FK027, FR17027, CL284635): spectrum against recent clinical isolates, comparative antimicrobial activity, beta-lactamase stability, and preliminary susceptibility testing criteria.

Cefixime, a new orally absorbed cephalosporin, was compared by in vitro testing with other oral beta-lactams, including cephalexin, cefaclor, cefuroxime, amoxicillin, and amoxicillin + clavulanate. Enterobacteriaceae were inhibited by lower concentrations of cefixime than any of the reference drugs; 90% and 95% were inhibited by less than or equal to 1.0 and less than or equal to 8.0 micrograms/ml, respectively. Cefixime was the least active among these drugs against staphylococci, with only 31% of 1106 strains inhibited by less than or equal to 8.0 micrograms/ml and less than 1% by less than or equal to 1.0 microgram/ml. Enterococci and pseudomonads were not susceptible to any of the drugs tested. Penicillin-resistant pneumococci were relatively resistant to cefixime, but penicillin-susceptible pneumococci were very susceptible to cefixime. Other streptococci were generally susceptible to all compounds tested, with relative activities of amoxicillin greater than cefaclor and cefuroxime greater than cefixime greater than cephalexin. Cefixime was inactive against Bacteroides species. A slight inoculum effect occurred with cefixime with inocolum concentrations varying from 10(5) to 10(6) colony forming units per milliliter, but this was more marked at 10(7) colony forming units per milliliter. Cefixime was resistant to hydrolysis by seven common beta-lactamases. It inhibited the hydrolysis of nitrocefin only by type 1 cephalosporinases. The disk diffusion zone diameter breakpoints for the 30-micrograms cefixime disk were determined by regression analysis to be greater than or equal to 27 mm (susceptible) and less than or equal to 23 mm (resistant), respectively corresponding to minimal inhibitory concentration breakpoints of less than or equal to 1.0 and greater than or equal to 4.0 micrograms/ml. Because of the high interpretive error rate (13.8%) and the occurrence of these breakpoints on the parabolic portion of the regression curve, we recommend further evaluation of cefixime disks with lower potencies.

CI-934, a new difluoroquinolone: in vitro antibacterial activity and proposed disk diffusion test interpretive criteria.

The susceptibility of 7,763 clinical isolates at four medical centers to CI-934 and three comparative quinolones was tested. CI-934 was the most active compound against Gram-positive isolates, such as staphylococci (MIC 90 = 0.25 microgram/ml), and enterococci (MIC 90 = 0.5 microgram/ml). CI-934 was the least active of these drugs against Pseudomonas spp. (MIC 90 = greater than 8.0 micrograms/ml). Against all other organisms CI-934 was very effective, being most comparable with enoxacin. With a selected group of isolates, CI-934 demonstrated high activity against Haemophilus influenzae (MIC 90 = 0.06 microgram/ml), Neisseria meningitidis and N. gonorrhoeae (MIC 90 = 0.13 microgram/ml), Listeria monocytogenes (MIC 90 = 1.0 microgram/ml), methicillin-resistant staphylococci (MIC 90 = 0.13 microgram/ml), and modest activity against anaerobes. Disk diffusion susceptibility testing of CI-934 was evaluated using 3-, 5-, and 10-micrograms disks. Because of its lower interpretive error rate, the 3-micrograms disk is tentatively recommended. With less than or equal to 2 micrograms/ml and greater than 4 micrograms/ml as the susceptible and resistant MIC breakpoints, the corresponding 3-micrograms disk zone diameters breakpoints are greater than or equal to 15 mm and less than or equal to 11 mm. Because most isolates of Pseudomonas spp. are not susceptible to CI-934 and the zone size interpretive error rate is particularly high (33%) with Pseudomonas spp., we suggest that isolates of this genus not be tested for clinical purposes.

Antimicrobial activity of Ro 19-5247 (T-2525), a new oral cephalosporin, tested against 7,745 recent clinical isolates.

The susceptibility testing of 7,745 recent clinical isolates from four medical centers showed Ro 19-5247 to be eight- to greater than 64-fold more active than cephalexin against the Enterobacteriaceae. Ro 19-5247 was comparable with cephalexin in anti-staphylococcal activity (MIC50, 4.0 micrograms/ml) and fourfold more active than cefixime. None of the oral cephalosporins were effective (MIC50, greater than 32 micrograms/ml) against enterococci, Pseudomonas aeruginosa and P. maltophilia. beta-lactamase hydrolysis experiments failed to demonstrate significant Ro 19-5247 inactivation by ten commonly encountered chromosomal- or plasmid-mediated enzymes (P99, K1, K14, TEM, CARB, OXA).

Interpretive criteria, quality control guidelines, and drug stability studies for susceptibility testing of cefotaxime, cefoxitin, ceftazidime, and cefuroxime against Neisseria gonorrhoeae.

Cefotaxime, cefoxitin, ceftazidime, and cefuroxime were tested in a multicenter study to establish susceptibility testing criteria and quality control guidelines for Neisseria gonorrhoeae. Interpretive criteria were established by using triplicate testing of at least 100 gonococcal strains having various susceptibility patterns to currently utilized drug regimens. Only a susceptible category was proposed for cefotaxime and ceftazidime (greater than or equal to 31 mm and less than or equal to 0.5 micrograms/ml) because of rare resistant isolates. The other interpretive crtieria were cefoxitin susceptible, greater than or equal to 28 mm (less than or equal to 2 micrograms/ml); intermediate, 24-27 mm (4 micrograms/ml), and resistant, less than or equal to 23 mm (greater than or equal to 8 micrograms/ml); cefuroxime-susceptible, greater than or equal to 31 mm (less than or equal to 1 micrograms/ml); moderately susceptible, 26-30 mm (2 micrograms/ml); and resistant, less than or equal to 25 mm (greater than or equal to 4 micrograms/ml). Quality control criteria were established by using multiple agar lots, three disk lots, and a number of replicates consistent with National Committee for Clinical Laboratory Standards M23-T guidelines.

RO 23-6240 (AM-833), a new fluoroquinolone: in vitro antimicrobial activity and tentative disk diffusion interpretive criteria.

The susceptibility of over 7000 recent clinical bacterial isolates to RO 23-6240, a new trifluorinated quinolone, was determined at four medical centers. Over 99% of Enterobacteriaceae and 97% of staphylococci were inhibited by less than or equal to 2.0 micrograms/ml of RO 23-6240. Only 71% of Pseudomonas spp. were inhibited by this concentration. Streptococci and enterococci were resistant to RO 23-6240. Clinical isolates of Haemophilus spp., pathogenic Neisseria spp., and Branhamella catarrhalis were inhibited by less than or equal to 0.25 micrograms/ml of RO 23-6240. This drug's antibacterial activity was comparable with that of enoxacin and norfloxacin, but was less than that of ciprofloxacin against most species. Using less than or equal to 2.0 micrograms/ml and greater than or equal to 8.0 micrograms/ml as the susceptible and resistant MIC breakpoints for RO 23-6240, the regression analysis-derived disk diffusion zone diameter breakpoints for the 5 micrograms disk are: Susceptible greater than or equal to 19 mm intermediate 16-18 mm, and resistant less than or equal to 15 mm.

Quality control limits for microdilution susceptibility tests with aztreonam, imipenem, ceftriaxone, ceftazidime, ceftizoxime, cefuroxime, and cefonicid.

A six-laboratory collaborative study was performed in order to define quality control limits for microdilution tests with seven new beta-lactams (aztreonam, imipenem, ceftriaxone, ceftazidime, ceftizoxime, cefuroxime, and cefonicid). Four standard control strains were tested and the expected MIC limits for each of the appropriate drug-microorganism combinations were defined.

Ticarcillin: a collaborative in vitro comparison with carbenicillin against over 9,000 clinical bacterial isolates.

The minimal inhibitory concentrations of ticarcillin and carbenicillin were determined for 9,236 clinical bacterial isolates by the broth microdilution method at four participating laboratories. Ticarcillin showed significantly increased activity against Klebsiella pneumoniae (P less than .001), Pseudomonas aeruginosa (P less than .001) and Aeromonas hydrophilia (P less than .005) when compared to carbenicillin, but no signifcant differences were observed against other gram-negative organisms. Ticarcillin was consistently less active against the gram-positive cocci, and these differences were significant for Staphylococcus aureus (P less than .001), Streptococcus agalactiae (P less than .001), Staphylococcus epidermidis (P less than .001) and Streptococcus viridans (P less than .005). Significant regional and institutional differences in susceptibility to the two drugs were observed for several species, including common nosocomial pathogens such as S. aureus, P. aeruginosa, K. pneumoniae and Escherichia coli.

BL-S786, a new parenteral cephalosporin. I. A collaborative in vitro susceptibility comparison to cephalothin against 5,762 clinical bacterial isolates.

The in vitro activity of Compound BL-S786 was compared with that of cephalothin against 5,762 clinical isolates by the microdilution broth method. BL-S786 demonstrated a broader spectrum and a significantly lower MIC against the Enterobacteriaceae. Although greater susceptibility to BL-S786 than to cephalothin was exhibited by Serratia marcescens, Proteus morganii and Proteus vulgaris, these three species were generally resistant to both drugs. By contrast, the staphylococci were significantly more susceptible to cephalothin than to BL-S786. Resistance to both drugs was demonstrated by Pseudomonas aeruginosa and other pseudomonads, enterococci and Bacteroides fragilis.

Piperacillin (T-1220), a new semisynthetic penicillin: in vitro antimicrobial activity comparison with carbenicillin, ticarcillin, ampicillin, cephalothin, cefamandole and cefoxitin.

Piperacillin (T-1220) is a new semisynthetic penicillin with an unusually broad spectrum of antimicrobial activity. In vitro comparisons of this drug with 6 other beta-lactam antimicrobics (ticarcillin, carbenicillin, ampicillin, cephalothin, cefamandole and cefoxitin) were conducted. These included minimal inhibitory concentrations (MIC) against 394 bacterial isolates, the minimal lethal concentrations (MLC) against 79 of those, as well as the effect of inoculum size on the MIC and MLC of the drugs. Piperacillin had significantly greater activity than did the other penicillins against Pseudomonas species and Klebsiella pneumoniae. Against P. aeruginosa piperacillin was 8- and 16-fold more active than ticarcillin and carbenicillin, respectively. The MLC of piperacillin rarely differed from the MIC by more than one log2 dilutions except against P. aeruginosa in which the MLC was 4-fold greater or more than the MIC of 45% of isolates tested. Ticarcillin, carbenicillin and cefoxitin showed minimal inoculum size effects. Cefamandole results showed the greatest inoculum size variation with 55% and 37% of isolates showing an 8-fold increase in MIC and MLC respectively by increasing inoculum from 10(5) to 10(7) CFU/ml. Piperacillin was intermediately effected having 25% of strains greater than 8-fold increase in MIC.