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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

Koserella trabulsii, a new genus and species of Enterobacteriaceae formerly known as Enteric Group 45.

The name Koserella trabulsii is proposed for a group of Enterobacteriaceae formerly called Enteric Group 45. This group consists of 12 strains that were originally identified as atypical Hafnia alvei. K. trabulsii strains were negative for indole production, Voges-Proskauer, H2S production, urea hydrolysis, phenylalanine deaminase, and acid production from glycerol, lactose, sucrose, and D-sorbitol; they were positive for methyl red, citrate (Simmons), lysine and ornithine decarboxylases, arginine dihydrolase (negative in 1 to 2 days and positive in 3 to 7 days), and acid production from cellobiose and melibiose; and they were resistant to the Hafnia-specific bacteriophage of Guinée and Valkenburg. They were tested for DNA relatedness by the hydroxyapatite method with 32PO4-labeled DNA from the designated type strain (CDC 3349-72, ATCC 35313). The 12 strains were 87 to 99% related in 60 degrees C reactions. Relatedness of K. trabulsii to 71 DNA hybridization reference strains of representative species of Enterobacteriaceae was 4 to 37%. It was 15 to 16% related to H. alvei. All strains were susceptible to nalidixic acid, sulfadiazine, gentamicin, kanamycin, and chloramphenicol, and 83% were susceptible to nalidixic acid, sulfadiazine, gentamicin, kanamycin, and chloramphenicol, and 83% were susceptible to tetracycline. Most of the strains were resistant or intermediate to penicillin, ampicillin, carbenicillin, colistin, and cephalothin. Five of the strains were isolated from wounds, three were from the respiratory tract, and one each was from a stool, knee fluid, water, and an unknown source. The clinical significance of this organism is not known; therefore, future studies should focus on its isolation and its relationship to human disease.

Adult↗
PubMed1985

Leminorella, a new genus of Enterobacteriaceae: identification of Leminorella grimontii sp. nov. and Leminorella richardii sp. nov. found in clinical specimens.

Leminorella is proposed as a new genus for the group of Enterobacteriaceae formerly known as Enteric Group 57. Strains of Leminorella gave positive tests for H2S production, acid production from L-arabinose and D-xylose, and tyrosine clearing; they were negative for indole production, Voges-Proskauer, urea hydrolysis, phenylalanine deaminase, motility, gelatin liquefaction, lysine and ornithine decarboxylases, arginine dihydrolase, growth in KCN, and acid production from adonitol, D-arabitol, cellobiose, erythritol, D-galactose, myo-inositol, lactose, maltose, D-mannitol, D-mannose, melibiose, alpha-CH3-glucoside, raffinose, L-rhamnose, salicin, D-sorbitol, sucrose, and trehalose. By DNA hybridization, strains of Leminorella were only 3 to 16% related to other Enterobacteriaceae and were divided into three groups. Leminorella grimontii is proposed as the type species for the genus and strain CDC 1944-81, ATCC 33999, is designated as the type strain. There were four strains of L. grimontii from stool specimens and two from urine specimens. L. richardii is proposed as the name for the second species (type strain, CDC 0978-82, ATCC 33998). All four L. richardii strains were from stool specimens. L. grimontii can be distinguished from L. richardii because it produces gas from glucose (100%) and acid from dulcitol (83%) and is methyl red positive (100%). One strain, CDC 3346-72, was more related to L. grimontii by DNA hybridization than to L. richardii, but the lower relatedness to both of these species indicated that it may be a third species. Biochemically it could not be distinguished from L. grimontii. All Leminorella strains were resistant (no zone of inhibition) to ampicillin, carbenicillin, and cephalothin. Some of the Leminorella strains were sent to us for Salmonella serotyping, and two reacted weakly in Salmonella antisera. The clinical significance of Leminorella is unknown.

DNA, Bacterial↗
PubMed1985

Comparison of the API rapid E four-hour system with the API 20E overnight system for the identification of routine clinical isolates of the family Enterobacteriaceae.

Four hundred forty-one clinical isolates of the family Enterobacteriaceae were identified in parallel by using the API Rapid E 4-h and the API 20E overnight procedures (Analytab Products, Plainview, N.Y.). The results obtained by using the API Rapid E were compared with those obtained by using the API 20E. Discrepancies were resolved by using standard biochemicals. The API 20E identified 98.9% (436 of 441) of the isolates without the use of additional biochemicals and was found to be correct in each case of a discrepancy among the 436 isolates. The API Rapid E gave the same identification as the API 20E for 94.0% (410 of 436) of the isolates, misidentified 3.0% (13 of 436), and gave a correct but low-selectivity answer for the remaining 3.0% (13 of 436). The API Rapid E is a suitable alternative for the rapid identification of the Enterobacteriaceae.

Bacteriological Techniques↗
PubMed1987

Cobas-Bact system for identification of members of the family Enterobacteriaceae in 4 h 20 min.

The Cobas-Bact (Roche Diagnostics, Basel, Switzerland) new rotor for the identification (ID) in 4 h 20 min of 33 members of the family Enterobacteriaceae to genus and species level was evaluated by testing 444 strains of which 398 belonged to common species and 46 belonged to rare species of Enterobacteriaceae. Each strain was identified by the API 20E system (Analytab Products, Plainview, N.Y.), and additional discriminating tests were set up if necessary. Only first-choice ID were considered in this study and were classified either as high-confidence ID (normalized likelihood, greater than or equal to 80%) or as low-confidence ID (normalized likelihood, less than 80%) requiring additional tests for confirmation. The data were analyzed by two versions of Cobas-Bact software. With the first version of the software (SW8446), the overall accuracy of Cobas-Bact was 95.5% (424 correct ID of 444). When restricted to high-confidence ID it rose to 99.4% (350 of 352) for the common species and 96.9% (31 of 32) for the rare species. Only three strains of the high-confidence group were misidentified. Sixty ID were considered unacceptable because of their low confidence. Using the first software version (SW8446) they represented 12% (46 of 398) of the common species (17 typical strains, 10 Shigella species, 10 inactive Escherichia coli strains, and 9 rare biotypes) and 30% (14 of 46) of the rare species. The same data analyzed by the new version (SW8524) of the Cobas-Bact software resulted in an overall accuracy of 93.9% (417 correct ID). The number of high-confidence ID rose to 401, of which 392 (97.7%) were accurate. The decrease in low-confidence ID (43 versus 60) was mainly due to the Shigella species. In conclusion the accuracy of Cobas-Bact identification system was very good when restricted to high-confidence ID. The Cobas-Bact performance for rare species ID was poorer, but the small number of strains tested does not allow definitive conclusions.

Bacteriological Techniques↗
PubMed1988

Phosphatase activity is a constant feature of all isolates of all major species of the family Enterobacteriaceae.

In this study we evaluated phosphatase activity in members of the family Enterobacteriaceae by conventional methods and by a novel method. The novel method is based on the formation of bright-green-strained colonies by phosphatase-positive, but not phosphatase-negative, strains in the presence of a phosphate substrate, such as phenolphthalein monophosphate or 6-benzoylnaphthyl phosphate (6-BNP), and methyl green. A total of 1,055 strains belonging to 65 different species of Enterobacteriaceae were tested for green staining of the colonies in the presence of methyl green and either phenolphthalein monophosphate or 6-BNP and for phosphatase activity by three different conventional methods. With the sole exception of one Leminorella richardii type strain, all isolates of all of the species formed green-stained colonies in the presence of the substrate 6-BNP. All strains were phosphatase positive by all of the conventional methods.

Enterobacteriaceae↗
PubMed1988

Quality of powdered substitutes for breast milk with regard to members of the family Enterobacteriaceae.

Members of the family Enterobacteriaceae were cultured from 52.5% of 141 milk substitute infant formulas which were obtained in 35 countries. The concentration did not exceed a level of 1 CFU/g in any product. The species which were isolated most frequently were Enterobacter agglomerans, cloacae, Enterobacter sakazakii, and Klebsiella pneumoniae. If infections due to these organisms occur, it can be useful to include a check of the hygienic precautions which are taken during the preparation and storage of the formula. Milk powders without members of the Enterobacteriaceae might offer extra protection to the newborn if some multiplication does occur in the formula.

Enterobacter↗
PubMed1988

Evaluation of autoscan-4 for identification of members of the family Enterobacteriaceae.

A study was performed to compare the Autoscan-4 (MicroScan, Inc., Mahwah, N.J.) with conventional biochemical methods for identifying clinical isolates of the family Enterobacteriaceae. The Autoscan-4 yielded correct identification of 95.4% of the isolates at the species level and 98.4% at the genus level. Only one misidentification was observed. The identification of both common and less-common isolates of Enterobacteriaceae makes this system highly efficient.

Bacterial Typing Techniques↗
PubMed1990

Plasmid profiling of members of the family Enterobacteriaceae, lactobacilli, and bifidobacteria to study the transmission of bacteria from mother to infant.

Plasmid profiles of isolates of the family Enterobacteriaceae, lactobacilli, and bifidobacteria cultured from vaginal, oral, and rectal swabs collected from women soon after admission to a maternity hospital were compared with those of strains detected in the feces of their infants. Lactobacilli inhabiting the vaginas of the mothers did not appear to colonize the infant digestive tract, but evidence for the transmission of fecal isolates of the Enterobacteriaceae and bifidobacteria from mother to infant was obtained in four out of five cases. Many of the bifidobacteria isolated were plasmid-free but could be distinguished with biochemical profiles.

Bifidobacterium↗
PubMed1991

Description and evaluation of the semiautomated 4-hour ATB 32E method for identification of members of the family Enterobacteriaceae.

A study was performed to compare the rapid identification system ATB 32E (API-bioMérieux SA, La Balme-les-Grottes, France) with conventional biochemical methods for identifying 414 isolates of the family Enterobacteriaceae and the genus Aeromonas, mainly of clinical origin. Overall, 395 strains (95.4%) were correctly identified, with 48 (11.6%) requiring extra tests for complete identification. Ten strains (2.4%) were not identified, and nine (2.9%) were misidentified. The ATB 32E is a suitable alternative for rapid identification of members of the family Enterobacteriaceae.

Autoanalysis↗
PubMed1991

Glycosidase profiles of members of the family Enterobacteriaceae.

A total of 712 strains representing 47 taxa of the family Enterobacteriaceae were tested for the ability to hydrolyze 14 4-methylumbelliferyl (4-MU)-linked substrates within 3 h of incubation. In addition to the well-known differentiation potential of the hydrolysis of 4-MU-beta-D-galactopyranoside, 4-MU-beta-D-glucuronide, and 4-MU-beta-D-xylopyranoside, the hydrolysis of some other fluorogenic substrates (e.g., 4-MU-beta-D-fucopyranoside, 4-MU-N-acetyl-beta-D-galactosaminide, and 4-MU-alpha-D-galactopyranoside) can also be used for species differentiation within the family Enterobacteriaceae.

Enterobacteriaceae↗
PubMed1992

Evaluation of the autoSCAN-W/A system for rapid (2-hour) identification of members of the family Enterobacteriaceae.

We evaluated the ability of the Baxter autoSCAN-W/A System (MicroScan Division, Baxter Diagnostics, Inc., West Sacramento, Calif.) to use the rapid (2-h) gram-negative identification panel for accurate identification of members of the family Enterobacteriaceae. At 2 h, 353 of 467 (75.6%) strains in a challenge set of biochemically typical and atypical stock cultures were correctly identified to genus and species. Another 76 (16.3%) strains were correctly identified to genus and species after the performance of recommended additional biochemical testing. Thus, at 24 h, 91.9% of the 467 strains were correctly identified. Twenty-two strains (4.7%) were identified to the correct genus but the incorrect species, and 16 strains (3.4%) were misidentified. Of these 16 strains, 9 were incorrect at 2 h, and 7 were incorrect after the additional testing. Because the system is based on fluorogenic substrates, no conventional tests were readily available with which to compare aberrant reactions. These results suggest that the autoSCAN-W/A with its rapid gram-negative panels is acceptable for the identification of the Enterobacteriaceae in a clinical microbiology laboratory.

Bacterial Typing Techniques↗
PubMed1994

Evaluation of RapID onE system for identification of 379 strains in the family Enterobacteriaceae and oxidase-negative, gram-negative nonfermenters.

The ability of the RapID onE system (Innovative Diagnostic Systems, Inc., Norcross, Ga.) to identify 364 strains in the family Enterobacteriaceae and 15 oxidase-negative, gram-negative, nonfermentative rods was evaluated. Kits were inoculated with no. 2 McFarland standard suspensions, and reactions were interpreted after 4 h of incubation at 35 degrees C. Overall, the method correctly identified (to the species level or to the genus level for salmonellas and non-Shigella sonnei Shigella species) 363 strains (95.8%) without additional tests. For four strains (1.0%), additional tests were required to delineate the correct identification from a range of two or more possibilities; these included one Serratia liquefaciens (Serratia marcescens or Serratia liquefaciens), one Serratia rubidaea (Serratia rubidaea or Serratia odorifera), one Salmonella typhi (Leminorella richardii or Salmonella sp.) and one Yersinia enterocolitica (Yersinia frederiksenii, Yersinia intermedia, or Yersinia enterocolitica). Twelve strains (3.2%) were misidentified or yielded codes with no identification; these comprised one Citrobacter amalonaticus (no identification), three Enterobacter hormaechei (not in the RapID onE database; two Enterobacter amnigenus, one Enterobacter sp.), one Serratia liquefaciens (Enterobacter cloacae), one Serratia rubidaea (no identification), four Serratia fonticola (not in RapID onE database; two Enterobacter aerogenes, one Serratia marcescens, one not identified), one Proteus mirabilis (Proteus penneri), and one Proteus vulgaris (Providencia rustigianii). If the seven strains not included in the database had been excluded, correct identification rates would have risen to 97.6% without additional tests and 98.7% with additional tests, with misidentification rates dropping to 1.3%. The RapID onE system is easy to set up and the results are easy to read, and the system provides an accurate, nonautomated commercially available method for the same-day identification of members of the family Enterobacteriaceae and oxidase-negative, gram-negative nonfermenters.

Bacteriological Techniques↗
PubMed1995

Evaluation of new medium with chromogenic substrates for members of the family Enterobacteriaceae in urine samples.

A new medium containing 5-bromo-4-chloro-3-indolyl-beta-D-glucuronide cyclohexylammonium salt (Glu agar) for Escherichia coli and a new medium containing 5-bromo-3-indolyl-beta-D-galactoside (Gal agar) for beta-galactosidase-positive members of the family Enterobacteriaceae were compared with MacConkey agar in a diagnostic trial with 3,562 urine specimens. The isolation rates of E. coli and beta-galactosidase-positive Enterobacteriaceae were increased 8.4 and 19.5%, respectively. The sensitivities and specificities of Glu agar and Gal agar were 98.5 and 100% and 99.2 and 99.5%, respectively.

Bacterial Typing Techniques↗
PubMed1997

Multicenter laboratory evaluation of the bioMérieux Vitek antimicrobial susceptibility testing system with 11 antimicrobial agents versus members of the family Enterobacteriaceae and Pseudomonas aeruginosa.

A four-center study in which a total of 1,082 recent clinical isolates of members of the family Enterobacteriaceae and Pseudomonas aeruginosa were examined versus 11 antimicrobial agents with the bioMérieux Vitek susceptibility test system (Hazelwood, Mo.) and the GNS-F6 card was conducted. In addition, a challenge set consisting of the same 200 organisms was examined in each of the four participating laboratories. Results obtained with the Vitek system were compared to MICs determined by a standardized broth microdilution method. For purposes of comparison, susceptibility categories (susceptible, intermediate, or resistant) were assigned on the basis of the results of both methods. The result of the broth microdilution test was considered definitive. The total category error rate with the Vitek system and the recent clinical isolates (11,902 organism-antimicrobial comparisons) was 4.5%, i.e., 1.7% very major errors, 0.9% major errors, and 1.9% minor errors. The total category error rate calculated from tests performed with the challenge set (i.e., 8,800 organism-antimicrobial comparisons) was 5.9%, i.e., 2.2% very major errors, 1.1% major errors, and 2.6% minor errors. Very major error rates higher than the totals were noted with Enterobacter cloacae versus ampicillin-sulbactam, aztreonam, ticarcillin, and ticarcillin-clavulanate and with P. aeruginosa versus mezlocillin, ticarcillin, and ticarcillin-clavulanate. Major error rates higher than the averages were observed with Proteus mirabilis versus imipenem and with Klebsiella pneumoniae versus ofloxacin. Excellent overall interlaboratory reproducibility was observed with the Vitek system. The importance of inoculum size as a primary determinant in the accuracy of susceptibility test results with the Vitek system was clearly demonstrated in this study. Specifically, when an inoculum density fourfold higher than that recommended by the manufacturer was used, high rates of false resistance results were obtained with cell wall-active antimicrobial agents versus both the Enterobacteriaceae and P. aeruginosa.

Anti-Bacterial Agents↗
PubMed2003

Occurrence and phenotypic characteristics of extended-spectrum beta-lactamases among members of the family Enterobacteriaceae at the Tel-Aviv Medical Center (Israel) and evaluation of diagnostic tests.

We assessed the prevalence and phenotypic characteristics of extended-spectrum beta-lactamase (ESBL) producers among cefuroxime-resistant (CXM-R) (MIC > or = 32 micro g/ml) members of the family Enterobacteriaceae in our institution. The 438 CXM-R clinical isolates obtained from nonurine sources among inpatients were screened. ESBL production was confirmed by disk diffusion assay using cefpodoxime (CPD), cefotaxime (CTX), and ceftazidime (CTZ) with and without clavulanate (CLAV). A difference of > or =5 mm in the size of the zone of inhibition in the presence of CLAV for at least one of the agents was considered representative of the ESBL phenotype: 186 isolates (42.5%) were confirmed as ESBL producers. The isolates tested and the rates of ESBL producers were as follows: Klebsiella spp. (n = 81), 79%; Proteus spp. (n = 58), 62%; Escherichia coli (n = 64), 53%; Enterobacter spp. (n = 69), 42%; Serratia spp. (n = 70), 14%; Citrobacter spp. (n = 25), 24%; Providencia spp. (n = 21), 24%; Morganella spp. (n = 41), 5%; and Kluyvera (n = 3), 0%. The overall sensitivity of isolated ESBL confirmatory tests was 79% for CPD-CLAV, 66% for CTZ-CLAV, and 91% for CTX-CLAV. Sensitivities of CTZ-CLAV confirmatory tests for Klebsiella spp., Proteus spp., E. coli, and Enterobacter spp. were 84, 22, 76, and 62%, respectively, and those for CTX-CLAV were 95, 97, 94, and 83%, respectively. They were 90% for CPD-CLAV and CTZ-CLAV, 95% for CPD-CLAV and CTX-CLAV, and 100% for CTZ-CLAV and CTX-CLAV. ESBL production was highly prevalent among Enterobacteriaceae. Using resistance to CXM as an ESBL screening criterion is a suitable option in high-incidence areas where Klebsiella spp. are not the dominant ESBL producers. This screening criterion may simplify the screening test and improve its sensitivity, although at the price of testing more isolates. The CTX-CLAV combination confirmed ESBL producers better than the CTZ-CLAV combination, with sensitivity varying between species. Combined CTZ-CLAV and CTX-CLAV testing detected all these strains; CPD-CLAV provided no additional benefit.

Anti-Bacterial Agents↗
PubMed2003

Modification of the double-disk test for detection of enterobacteriaceae producing extended-spectrum and AmpC beta-lactamases.

Detection of extended-spectrum beta-lactamases (ESBLs) in AmpC-producing Enterobacteriaceae is problematic. A modification of the double-disk test (MDDT) has been developed for successful detection of ESBLs in gram-negative bacilli producing well-characterized beta-lactamases as well as 212 clinical isolates of Enterobacter cloacae, Enterobacter aerogenes, Serratia marcescens, and Citrobacter freundii. MDDT accurately differentiated between ESBL producers and derepressed chromosomal AmpC mutants. MDDT provides a cost-effective alternative approach for clinical microbiology laboratories for routine susceptibility testing with simultaneous detection of ESBLs in enterobacteriaceae.

Bacterial Proteins↗
PubMed1978

Comparison of micro-ID, API 20E, and conventional media systems in identification of Enterobacteriaceae.

The Micro-ID, a new identification kit for Enterobacteriaceae, consists of 15 biochemical tests, with substrates and reagents impregnated in filter paper disks. A 0.2-ml amount of an organism suspension equal to a 0.5 McFarland standard is pipetted into each of the compartments. After 4 h of incubation and addition of potassium-hydroxide (KOH) to the Voges-Proskauer test, the color reactions are read according to the recommendations of the manufacturer. A five-digit octal code number is derived from each set of reactions from which an identification is derived by using a code book. In a single-blind, comparative study of the Micro-ID system with the API 20E system (Analytab Products Inc.) and conventional biochemical tube media, we found that the Micro-ID and the API 20E systems gave a 90% identification correlation when each was compared with the conventional tube media. A comparison of all three systems gave an 82% overall identification correlation. When the common tests of Micro-ID and API 20E were compared with conventional tube media, we found that the tests in the Micro-ID performed as well as or better than those of the API 20E. Certain groups of organisms, i.e., Citrobacter, Enterobacter, Proteus, Salmonella, and Serratia genera, were found to give low correlation on certain common tests. When using primary isolation MacConkey plates from the clinical laboratory, only 74% of the plates with Enterobacteriaceae had sufficient numbers of colonies of each enteric organism to produce the 0.5 McFarland inoculum density required. Problems concerning the misidentification of some organisms are discussed.

Bacteriological Techniques↗