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PubMed1983

Evaluation of the Sensititre system for identification of Enterobacteriaceae.

The Sensititre identification system (Seward Laboratory/GIBCO Laboratories) consists of a microplate containing a pattern of 24 biochemicals repeated four times together with an automatic inoculation device and a microcomputer-assisted data interpretation component. A total of 1,415 isolates of Enterobacteriaceae plus 6 isolates of other glucose-fermenting gram-negative bacilli were tested in three hospital laboratories in parallel with API 20E (Analytab Products). Discrepancies were resolved by conventional biochemical testing. Sensititre yielded correct identifications at the species level with 94.6% of the isolates and at the genus level with an additional 1.9%. API 20E yielded correct species identification with 91.1% and genus only identification with an additional 6.7% of the isolates. For the routine identification of clinical Enterobacteriaceae isolates, the Sensititre system compares favorably with API 20E and offers clinical laboratories the economy of a microtiter plate system as well as the benefit of a microcomputer capable of other microbiological and data management applications.

Bacteriological Techniques↗
PubMed1975

Evaluation of the enteric analyzer for identification of Enterobacteriaceae.

The reliability of the Enteric Analyzer for identification of Enterobacteriaceae was evaluated using biochemical results previously obtained for 291 organisms with the conventional, R/B, and Minitek systems. The instrument correctly identified 77.3% of the organisms using conventional system results, 74.2% using R/B results, and 60.5% using Minitek results. The low rate of identification with the conventional system occurs primarily because the instrument is not programmed to consider delayed biochemical reactions. The arbitrary use of 90% and 99% probabilities for test reactions also contributes to a lower percentage of identification. The Enteric Analyzer does not replace the judgment of experienced personnel in the identification of atypical bacteria, but it may prove helpful in speeding up final computer identification of typical microorganisms.

Classification↗
PubMed1984

Reproducibility of the MS-2 system for identification of members of the family Enterobacteriaceae: a collaborative study with blindly assigned reference stains.

The reproducibility of identification and biochemical reactions for five different reference organisms of Enterobacteriaceae; Proteus vulgaris, Klebsiella pneumoniae, Escherichia coli, Serratia marcescens, and Enterobacter cloacae, were evaluated using the updated MS-2 system software (Abbott Laboratories, Diagnostic Division, Irving, Tex.) in a collaborative study involving 11 laboratories. When a total of 220 randomly coded test organisms were blindly examined, the MS-2 system correctly identified 92.7 and 86.8% for over 80 and 90% probability identification, respectively. Four organisms, P. vulgaris, K. pneumoniae, E. coli, and S. marcescens, were correctly identified in all laboratories with high probability, but 9 of 44 tests of Enterobacter cloacae resulted in misidentifications or low-likelihood (less than 80%) identifications. Accuracy was directly related to level of experience and familiarity with the MS-2 system in the individual laboratories. Biochemical reactions varied among the identification trials, especially in the identification of S. marcescens and Enterobacter cloacae. Among a total of 44 subcultures for each organism, 10 different biochemical patterns for P. vulgaris, 6 for K. pneumoniae, 9 for E. coli, 15 for S. marcescens, and 14 for Enterobacter cloacae were obtained. The results indicate that the MS-2 system performs with high accuracy and reproducibility in identifying Enterobacteriaceae, except for Enterobacter cloacae.

Bacteriological Techniques↗
PubMed1985

Comparison of PRAS II, RapID ANA, and API 20A systems for identification of anaerobic bacteria.

This study evaluated the PRAS II, RapID ANA, and API 20A systems for the identification of anaerobic bacteria. A total of 80 isolates (68 fresh clinical isolates and 12 stock cultures) were examined and included 25 Bacteriodes spp., 7 Fusobacterium spp., 12 Clostridium spp., 2 Veillonella spp., 16 gram-positive cocci, and 18 gram-positive nonsporeforming bacilli. All isolates were initially identified by the procedures outlined in Holdeman et al. (ed.), Anaerobe Laboratory Manual, Virginia Polytechnic Institute and State University, Blacksburg, Va., 1977; identifications from the PRAS II, RapID ANA, and API 20A systems were compared with these initial identifications. If no supplemental tests were required, the RapID ANA and API 20A systems had incubation times of 4 and 24 h, respectively; the PRAS II system generally required 2 to 5 days of incubation, depending on the growth rate of the isolate. PRAS II identified 74% correct to species level, 14% correct to genus only, and 6% incorrect; 6% could not be identified. PRAS II data were reevaluated according to a revised data base that was provided after completion of the study; PRAS II (revised) identified 82% correct to species, 12% correct to genus only, and 6% incorrect. RapID ANA identified 62% correct to the species level, 28% correct to genus only, and 10% incorrect. API 20A identified 71% correct to the species level, 10% correct to genus only, and 3% incorrect; 16% could not identified. The API 20A is a more established system for identification of anaerobic bacteria; PRAS II and RapID ANA appear to be promising new methods for the identification of anaerobic bacteria.

Bacteria, Anaerobic↗
PubMed1985

Rapid species identification of group C streptococci isolated from horses.

Two commercial systems, the API 20S (Analytab Products, Plainview, N.Y.) and the Rapid Strep (API System S.A., Montalieu-Vercieu, France), were evaluated for ease of use and accuracy in the rapid identification of group C streptococci isolated from horses. A total of 85 Streptococcus isolates were tested, including S. equi (67 isolates), S. zooepidemicus (13 isolates), and S. equisimilis (5 isolates). All S. equi and S. zooepidemicus isolates were correctly identified within 24 h by the Rapid Strep system. Specific grouping sera was necessary to distinguish between S. equisimilis and group G or L strains. The API 20S system did not provide species identification of any of these isolates. An identification of randomly selected isolates to species level was performed by conventional methods and confirmed the identification derived through the Rapid Strep system. Our results indicate that the Rapid Strep system is a valuable aid for species identification of equine isolates of group C streptococci.

Animals↗
PubMed1985

Accuracy and reproducibility of a four-hour method for anaerobe identification.

In this study, we evaluated the ability of a 4-h enzyme assay kit system, the RapID ANA method (Innovative Diagnostic Systems, Inc., Atlanta, Ga.) to accurately and reproducibly identify a spectrum of clinically significant anaerobic bacteria in two separate institutions. Additional tests were performed as required. Of a total of 188 organisms tested at Hershey Medical Center (HMC), 86.2% were correctly identified to species level without additional tests, 5.9% required extra tests for correct identification, and 8.0% were misidentified. Of 53 strains tested at Johns Hopkins Hospital (JHH), 52.8% were correctly identified without extra tests, 28.3% required extra tests for correct identification, and 18.9% were misidentified. Of 21 organisms tested at both institutions, those tested at JHH required additional tests for correct identification in 38.1% of cases, compared with 9.5% at HMC. Misidentification rates were identical (9.5%) in both centers. Of strains tested at HMC only, 86.8% were correctly identified without extra tests, 5.4% were identified with additional tests, and 7.8% were misidentified: corresponding data for JHH were 53.1, 21.9, and 25.0%, respectively. Of 53 strains tested in triplicate at JHH, 56.7% yielded the same result on each occasion, 37.7% were identical in two of three tests, and 5.7% gave different results on each of three occasions. Discrepancies between identification rates at HMC and JHH may be explained by differences in species tested (more commonly encountered species were tested at HMC) and interpretation of reactions by the two different readers. The RapID ANA method has the potential for rapid identification of clinically isolated anaerobes; however, accuracy and reproducibility may vary as a function of the specific laboratory setting.

Bacteria, Anaerobic↗
PubMed1990

Reproducibility of API Staph-Ident system identifications of coagulase-negative staphylococci isolated from blood.

Fifty-five isolates of coagulase-negative staphylococci from blood were identified four times each by using the API Staph-Ident (API-SI) system to determine the identification reproducibility of the system. Identifications were determined by using both Version 1 and Version 2 of the API-SI Profile Index. The 75 to 100% reproducibility levels, including reproducible nonidentifications, were 98.2% for Version 1 and 96.4% for Version 2. Version 1 gave an identification for 92.5% of the isolates with a 75 to 100% reproducibility, while Version 2 gave an identification for 100% of the isolates with a 75 to 100% reproducibility. The reproducibility of the API-SI identifications of coagulase-negative staphylococci from blood was good.

Bacteriological Techniques↗
PubMed1990

Evaluation of MicroScan for identification of Enterococcus species.

Emerging drug resistance of the enterococci necessitates differentiation from group D streptococci and accurate species identification. MicroScan (Baxter Healthcare Corp., West Sacramento, Calif.) has recently developed a microdilution system for identification and antibiotic susceptibility testing of gram-positive cocci. To evaluate the ability of this system to identify Enterococcus species, 100 isolate identified as enterococci by MicroScan were tested by conventional media and 60 isolates of streptococci were tested by MicroScan. Incubation times for conventional and MicroScan methods were 96 and 18 to 24 h, respectively. For 94 strains of enterococci (77 Enterococcus faecalis, 14 Enterococcus faecium, and 3 Enterococcus durans), identification by conventional media and MicroScan agreed. Of the remaining six isolates, four were identified as E. faecalis and two were identified as E. durans by MicroScan, whereas by conventional media the four E. faecalis isolates were identified as Enterococcus solitarius and the two E. durans isolates were identified as Enterococcus hirae. None of the 60 streptococci were identified as enterococci. MicroScan is a reliable method for identification of the commonly encountered enterococcal species E. faecalis and E. faecium; however, modifications of the system are necessary for identification of other Enterococcus species.

Autoanalysis↗
PubMed1990

Evaluation of the ATB 32 A system for identification of anaerobic bacteria isolated from clinical specimens.

A new miniaturized 4-h method for the identification of anaerobic bacteria, ATB 32 A (API System SA, Montalieu Vercieu, France), was evaluated against conventional methods of identification. The evaluation was done by using 260 recent clinical isolates and 21 reference strains of anaerobic bacteria. All reference strains were correctly identified and did not figure in the detailed analysis. Of the 140 gram-negative bacilli, 90.6% of Bacteroides spp. and 95.5% of Fusobacterium spp. were correctly identified to the species level, with an additional 8.4% of the Bacteroides spp. being identified to the genus level. Clostridia were correctly identified in 85.9% of cases, with an additional 9.9% being identified to the genus level. Peptostreptococci were correctly identified in 91.6% of cases. The 4 strains of Actinomyces spp. were all identified correctly, as were 10 of the 11 strains of Propionibacterium spp. A total of 3.1% of strains were not identified by ATB 32 A, while for 1.9% of strains, completely false identifications were obtained. Estimation of the individual preformed enzyme results may pose problems, although these decrease with familiarity with the system. With certain enzyme profiles, additional testing was necessary to arrive at an identification; however, there was no requirement for gas-liquid chromatography. If certain additions are made to the data base and the difficulties of determination of organisms to the species level among the non-Bacteroides fragilis (sensu stricto) members of the B. fragilis group can be reduced, this system holds promise as a reliable standardized alternative for the identification of anaerobic bacteria in clinical laboratories.

Bacteria, Anaerobic↗
PubMed1991

Evaluation of practical chromatographic procedures for identification of clinical isolates of mycobacteria.

After experimental conditions were established, 366 strains of mycobacteria belonging to 23 different species were studied for fatty acids, secondary alcohols, and mycolic acid cleavage products by capillary gas-liquid chromatography. Additionally, the mycolic acid pattern was studied by thin-layer chromatography. Capillary gas-liquid chromatography allowed direct identification of the following Mycobacterium spp.: M. kansasii, M. marinum, M. szulgai, M. xenopi, M. malmoense, and M. gordonae. The patterns of mycolic acid methyl esters recorded for the test strains of M. chelonae and M. agri may be of value in the identification of these species. Moreover, the combined use of the two chromatographic techniques provided precise identification of the M. tuberculosis complex, M. simiae, M. fallax, M. triviale, and M. chelonae-like organisms. A minimal set of biochemical tests is usually required to obtain identification to the species level when chromatographic procedures alone are not sufficient. Under the reported experimental conditions, thin-layer chromatography and capillary gas-liquid chromatography are rapid and very useful techniques for the identification of mycobacteria.

Alcohols↗
PubMed1991

Evaluation of the Baxter-MicroScan 4-hour enzyme-based yeast identification system.

A new 4-h Yeast Identification Panel (YIP; Baxter-MicroScan, W. Sacramento, Calif.) was compared with the API 20C Yeast Identification System (Analytab Products, Inc., Plainview, N.Y.) in the identification of recent clinical yeast isolates. The YIP had a 94% correlation (288 of 306) in identifying 22 species within the genera Candida, Hansenula, Pichia, Rhodotorula, Saccharomyces, and Torulopsis. Correlation dropped to 65% for those species within the genera of slower growing yeasts, i.e., Blastoschizomyces spp., Crpytococcus spp., Geotrichum spp., Hyphopichia spp., Phaeococcomyces spp., Prototheca spp., and Trichosporon spp. Overall correlation with the API 20C was 92% (365 of 401) for those taxa included in the data base and 85% (373 of 437) for all yeasts encountered in the study. There were 36 (8.2%) discrepant identifications, which were due in part to the limited data base. Expansion of the data base plus the easy inoculation, reading, and rapid results of the YIP should make it an excellent method for yeast identification.

Culture Media↗
PubMed1991

Preliminary evaluation of Biolog, a carbon source utilization method for bacterial identification.

The Biolog Identification System (Biolog, Inc., Hayward, Calif.) is a new bacterial identification method that establishes an identification based on the exchange of electrons generated during respiration, leading to a subsequent tetrazolium-based color change. This system tests the ability of a microorganism to oxidize a panel of 95 different carbon sources. We report on a preliminary investigation of the ability of the instrument to identify, using its computer-driven enzyme immunoassay reader, a diverse group of clinically relevant members of the family Enterobacteriaceae and gram-negative non-Enterobacteriaceae. The Biolog reported identifications (correct or incorrect) for 266 of 352 organisms tested (75.6%). Of the 266 identifications reported, 87.3% were correct at the genus level and 75.6% were correct at the species level at 24 h. In the total study of 352 strains, 46.6% were correct to the species level at 4 h and 57.1% were correct to the species level at 24 h. The error rate was 10.4% after 4 h and 9.6% after 24 h. The Biolog performed well with many genera, but problems were encountered with some strains of Klebsiella, Enterobacter, and Serratia. We found the system to be versatile and easy to use.

Bacteria↗
PubMed1992

Evaluation of the autoSCAN-W/A rapid system for identification and susceptibility testing of gram-negative fermentative bacilli.

The autoSCAN-Walk-Away (W/A) system for identification and susceptibility testing was evaluated for 400 gram-negative fermentative bacteria by using the API 20E (366 isolates) and/or tube biochemical tests as the reference identification system and a frozen microdilution MIC tray system for susceptibility testing. The W/A system performed well for identification of this group of organisms representing 14 genera and 30 species, showing a sensitivity of 96% and results available in 2 h. Of the 16 misidentifications, 6 were with Serratia liquefaciens. A total of 63 isolates (17%) required further tests to complete the identification, compared with 106 (29%) of the isolates which required additional tests for the API 20E identification. Approximately half (32) of the additional tests with the W/A system were required in order to separate Citrobacter diversus from C. amalonaticus. For susceptibility determinations, the W/A system demonstrated an overall agreement of 93% (4,102 determinations) with 40 major errors (0.98%). However, of the 906 resistant organism-drug combinations in the study, there were 115 very major errors, for a false-susceptibility rate of 12.7% of the resistance determinations. Among these very major errors, 80% occurred with piperacillin and the cephalosporins. The W/A system completed the MIC determinations in 7 h; however, the difficulty in detecting resistance with some antimicrobial agents limited the advantages of the rapid susceptibility testing.

Bacteriological Techniques↗
PubMed1992

Evaluation of the RapID ANA II and API ZYM systems for identification of Actinomyces species from clinical specimens.

Classification and identification of fermentative actinomycetes are labor-intensive and problematic. In this study, we evaluated the applicability and reliability of the RapID ANA II system (Innovative Diagnostic Systems, Inc., Atlanta, Ga.) and the discriminatory value of the API ZYM system (Societes Analytab Products Inc., La Balme Les Grottes, France) in the identification of Actinomyces-like bacteria by using conventional methods as a reference. Eighty-five strains, including 71 isolates from mixed anaerobic infections and 14 reference strains, were tested. The RapID ANA II system correctly identified all Actinomyces odontolyticus strains and 65% of Actinomyces israelii strains. All Arcanobacterium haemolyticum strains were misidentified as Actinomyces pyogenes. The most common isolates in the study were Actinomyces meyeri-like organisms, 84% of which, however, were aerotolerant. The identification of these aerotolerant strains thus remains unresolved and warrants further studies. New characteristics and changes to the conventional API ZYM enzyme profiles are suggested. The API ZYM enzyme profiles of A. odontolyticus and A. israelii were very similar, the only discriminating enzyme being alpha-fucosidase. In differentiation between A. pyogenes and Arcanobacterium haemolyticum, the production of beta-glucuronidase by the former and the production of acid phosphatase by the latter are suggested as new helpful characteristics for use in clinical laboratories. In summary, the RapID ANA II and API ZYM systems can be used as rapid preliminary methods in the identification of Actinomyces species but accurate identification requires supplementary conventional tests and gas-liquid chromatography.

Actinomyces↗
PubMed1992

Evaluation of the Autoscan Walkaway system for rapid identification and susceptibility testing of gram-negative bacilli.

We evaluated the performance of the Autoscan Walkaway (W/A) system (MicroScan, Sacramento, Calif.) in conjunction with the fluorometric Neg Combo panels for rapid identification and susceptibility testing of gram-negative bacilli. Fermentative and nonfermentative gram-negative bacilli were tested in parallel with the W/A system and the Cathra Repliscan replicator (C/R) system (Cathra, St. Paul, Minn.). Conventional biochemical testing and agar dilution testing were used to resolve the identification and susceptibility testing discrepancies. Of 495 clinical isolates tested, 445 (90%) were correctly identified by the W/A system and 483 (98%) were correctly identified by the C/R system. Repeat testing by using updated versions of the W/A system's computer identification software failed to demonstrate improved identification accuracy. For susceptibility testing, the W/A system demonstrated 5.6% total interpretative category errors, including only 0.9% major and very major errors. The comparative C/R system produced only 1% errors overall, including 0.2% major and very major errors. Although the W/A system is highly automated and is capable of producing results rapidly, our findings suggest that additional identification and susceptibility testing refinements are needed before the system will be suitable for routine use.

Anti-Bacterial Agents↗
PubMed1994

Comparison of updated Vitek Yeast Biochemical Card and API 20C yeast identification systems.

The updated Vitek Yeast Biochemical Card (YBC) was compared with the API 20C by using 409 germ tube-negative yeasts and Geotrichum spp. that were either clinical or proficiency sample isolates. The API 20C was the reference standard. The 409 isolates represented nine genera and 21 species. Morphology agars were inoculated and interpreted for each isolate. The API 20C identified 406 isolates (99.3%), while the Vitek YBC identified 367 (89.7%). Both systems identified the majority of yeasts after 24 h of incubation--73.4% were identified by the API 20C and 77.4% were identified by the Vitek YBC. The Vitek 24-h reading had some incorrect identifications. These included 14 isolates of Candida tropicalis that were identified as Candida parapsilosis (91 to 97% reliability) and 3 isolates of Candida krusei that were called Blastoschizomyces capitatus (Geotrichum capitatum), Candida rugosa, and Candida zeylanoides. In total, the Vitek YBC misidentified 30 isolates, while the API 20C misidentified 3 isolates. In addition, results for 14 isolates with the Vitek YBC were listed under the category "no identification." Morphology agars were required for identification with 89 isolates (21.9%) when the API 20C was used and with 50 isolates (12.6%) when the Vitek YBC was used. Apart from the price of the Vitek instrument, the API 20C costs $1.28 more per test than the Vitek YBC. Overall, the updated Vitek YBC compares favorably with the API 20C in the identification of common yeasts such as Torulopsis glabrata, C. parapsilosis, and Cryptococcus neoformans. However, problems were encountered with the Vitek system in the identification of C. tropicalis, C. krusei, Trichosporon spp., and some Cryptococcus spp. The routine use of morphology agars with either method is recommended.

Agar↗
PubMed1994

Gas-liquid chromatography of cellular fatty acids for identification of staphylococci.

A commercially available, computer-assisted microbial identification system (MIS) employs gas-liquid chromatographic analyses of bacterial fatty acids. The MIS was used to identify 470 isolates of Staphylococcus species. The accuracy of the MIS was compared with the accuracies of conventional methods. There was a complete agreement between the MIS and conventional methods in the identification of 413 (87.8%) strains. For 36 of 45 misidentified strains, the correct identification was listed by the MIS as a choice but not as the first choice. Twelve strains could not be matched. All strains of Staphylococcus cohnii, S. epidermidis, S. intermedius, S. lugdunensis, S. schleiferi, S. sciuri, S. simulans, and S. xylosus were correctly identified. Two species, S. hominis and S. saprophyticus, accounted for 52.6% (30 of 57) of the misidentifications. Seventy-eight organisms were retested. Identification of 73 organisms remained unchanged, and for five organisms, the second choice became first and vice versa. The overall performance of the MIS is acceptable, and the system can be used as an alternate identification method for staphylococci.

Chromatography, Gas↗
PubMed1994

Evaluation of RapiDEC Staph for identification of Staphylococcus aureus, Staphylococcus epidermidis, and Staphylococcus saprophyticus.

RapiDEC Staph is a test for presumptive identification of the principal human staphylococcal species, Staphylococcus aureus, S. epidermidis, and S. saprophyticus. The test includes control and test cupules for fluorogenic detection of coagulase and chromogenic substrates for alkaline phosphatase and beta-galactosidase. These tests identify S. aureus, S. epidermidis, and S. saprophyticus, respectively. Positive results with both chromogenic substrates provide a presumptive identification of S. xylosus or S. intermedius (S. xylosus-S. intermedius). Test cupules are inoculated with an organism suspension, and reactions are read after a 2-h incubation. RapiDEC-Staph was evaluated with 303 clinical and stock staphylococcal strains. Identifications were compared with those obtained by the tube coagulase test, a latex slide coagulase test (StaphAUREX), another commercial identification system (Staph-TRAC), and additional conventional tests. RapiDEC-Staph correctly identified 100% of 130 S. aureus strains, 70.3% of 74 S. epidermidis strains, and 81.3% of 32 S. saprophyticus strains. Four of five S. xylosus isolates were called S. xylosus-S. intermedius. Unidentified S. epidermidis and S. saprophyticus strains were called "Staphylococcus spp." Among the 62 other coagulase-negative staphylococci, 4 were misidentified as S. epidermidis and 7 were misidentified as S. saprophyticus. While the sensitivity and specificity of the fluorogenic coagulase test for S. aureus were 100%, failure to detect alkaline phosphatase activity in several S. epidermidis isolates resulted in fewer correct identifications by the RapiDEC-Staph test for this species.

Alkaline Phosphatase↗