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PubMed1966

Opsonizing and bactericidal activity of antibodies against common antigen of Enterobacteriaceae.

Domingue, Gerald J. (State University of New York at Buffalo, Buffalo, N.Y.), and Erwin Neter. Opsonizing and bactericidal activity of antibodies against common antigen of Enterobacteriaceae. J. Bacteriol. 91:129-133. 1966.-In addition to the well-known O, H, K, and Vi antigens, Enterobacteriaceae produce a common antigen, which was first identified by hemagglutination tests with Escherichia coli O14 antiserum. Studies on the biological significance of this antigen by in vitro phagocytic experiments have revealed that opsonization is markedly enhanced in the presence of the corresponding antibody, rabbit polymorphonuclear leukocytes, and normal rabbit serum. This effect is specific, because Pseudomonas aeruginosa, which is devoid of this antigen, is not opsonized under these conditions, and removal of the antibody by absorption markedly reduces the uptake of enteric bacteria containing the antigen. Opsonization thus represents another method for the study of this antigen-antibody system. Bactericidal tests have revealed that antibodies against this antigen, engendered in rabbits by different strains of enteric bacteria and various procedures, are bactericidal for E. coli O14 but not for other enteric bacteria, possibly due to previously demonstrated differences between the antigen moieties obtained from these microorganisms.

Antigens↗
PubMed1967

Release of surface enzymes in Enterobacteriaceae by osmotic shock.

The process of osmotic shock, which has been used to release degradative enzymes from Escherichia coli, can be applied successfully to other members of the Enterobacteriaceae. Cyclic phosphodiesterase (3'-nucleotidase), 5'-nucleotidase (diphosphate sugar hydrolase), acid hexose phosphatase, and acid phenyl phosphatase are released from Shigella, Enterobacter, Citrobacter, and Serratia strains. Some strains of Salmonella also release these enzymes. Members of Proteus and Providencia groups fail to release enzymes when subjected to osmotic shock and do not show a lag in regrowth, although they do release their acid-soluble nucleotide pools. In contrast to E. coli, release of enzymes from other members of the Enterobacteriaceae studied is affected by growth conditions and strain of organism. None of the organisms was as stable to osmotic shock in exponential phase of growth as was E. coli. Exponential-phase cells of Shigella, Enterobacter, and Citrobacter could be shocked only with 0.5 mm MgCl(2) to prevent irreparable damage to the cells. These observations suggest that this group of degradative enzymes is probably loosely bound to the cytoplasmic membrane through the mediation of divalent cations.

Acid Phosphatase↗
PubMed2006

Evaluation of the BD Phoenix automated microbiology system for identification and antimicrobial susceptibility testing of Enterobacteriaceae.

We evaluated the accuracy of the BD Phoenix system for the identification (ID) and antimicrobial susceptibility testing (AST) of 251 isolates of the family Enterobacteriaceae representing 31 species. Organisms were inoculated onto the Phoenix panel according to the manufacturer's instructions. The results from conventional biochemical tests were used for the reference method for ID. Agar dilution, performed according to the CLSI guidelines, was the reference AST method. Essential and categorical agreements were determined. The overall levels of agreement for the genus- and species-level identifications were 95.6% and 94.4%, respectively. Fourteen isolates were incorrectly identified by the Phoenix system; 10 of these were incorrectly identified to the species level. Three of these were Enterobacter (Pantoea) species and four of these were Shigella spp. misidentified as Escherichia coli. For AST results, the essential and categorical agreements were 98.7% and 97.9%, respectively. The very major error, major error, and minor error rates were 0.38%, 0.33%, and 1.8%, respectively. Six isolates (three E. coli isolates and three Klebsiella isolates) were extended-spectrum beta-lactamase producers. All six were flagged by the Phoenix system expert rules. The Phoenix system compares favorably to traditional methods for ID and AST of Enterobacteriaceae.

Anti-Bacterial Agents↗
PubMed1979

Rapid identification of Enterobacteriaceae with the micro-ID system versus API 20E and conventional media.

The Micro-ID system for rapid (4 h) identification of Enterobacteriaceae was evaluated by testing 433 enteric bacilli and 9 other gram-negative bacilli. Each isolate was identified with conventional tubed media and was also tested in the Micro-ID and API 20E systems. The overall accuracy of both systems was 97%. Micro-ID tests for the Voges-Proskauer reaction, indole and H2S production, and ornithine and lysine decarboxylase all demonstrated a 97 to 99% correlation with conventional methods. Only 86% of the Micro-ID urease tests agreed with Christenson urea agar. Two inoculum densities were tested in Micro-ID panels, with 157 stock cultures. Over 90% of the tests were unaffected by changes in inoculum density. Tests with four control strains suggested that the Micro-ID system was more reproducible when a light inoculum was used. The Micro-ID system was found to be a very convenient method for rapid, accurate, and precise identification of the Enterobacteriaceae.

Bacteriological Techniques↗
PubMed1979

Identification of Enterobacteriaceae in frozen microdilution trays prepared by Micro-Media Systems.

Frozen microdilution trays, with 20 different biochemical test media for identification of Enterobactericeae, were evaluated. The test panels were those provided by Micro-Media Systems (MMS), who also provided a code book for interpretation of the test results. The interpretations of the MMS test system and of the Analytab API 20E test system were compared with those obtained from tests with conventional media. Tests with 468 Enterobacteriaceae demonstrated nearly comparable results with the two commercial systems. About 6% (API) and 7% (MMS) of the isolates could not be identified without additional tests, and another 3% (MMS) to 4% (API) of the isolates gave test patterns that were not found in the code books. One to three percent of the isolates were misidentified. It was concluded that the MMS Enteric Quad Panels provide an efficient, relatively inexpensive, and reasonably accurate method for identification of the Enterobacteriaceae.

Bacteriological Techniques↗
PubMed1979

Rapid identification of Enterobacteriaceae from blood cultures with the Micro-ID system.

Micro-ID is a new test system designed to identify members of the family Enterobacteriaceae in 4 h. It consists of 15 biochemical tests on reagent-impregnated paper disks; each test is in its own compartment in a molded plastic tray. Based on the pattern of positive and negative biochemical reactions, a five-digit octal code number is calculated. A computer-generated identification manual accompanies the product, and for each octal code listed there is a numerical value that represents the unknown isolate's degree of fit to a typical organism (LFR), a second numerical value that represents its separation from other organisms (PNOR), and a verbal description of the quality of identification. Only one reagent is added to the system. Manufacturer's directions were modified in this laboratory to allow identification from a turbid blood culture bottle within 4 h. Based on 330 routine clinical cultures tested, there was a 96.1% agreement with conventional identification to the genus and species level: 1.2% yielded first two choices possible, with one being the correct choice; 1.2% provided a correct genus, but no species identification; and 1.5% produced an incorrect identification. The Micro-ID is an accurate, facile system for the rapid identification of Enterobacteriaceae from blood cultures.

Bacteriological Techniques↗
PubMed1981

Correlation studies of entero-set 20, API 20E and conventional media systems for Enterobacteriaceae identification.

The Entero-Set kit (Fisher Diagnostics) is a 20-biochemical-test system used in the identification of members of the Enterobacteriaceae. This kit was compared with the API 20E (Analytab Products) and conventional media systems, using 505 (303 stock and 202 clinical) strains of Enterobacteriaceae. When the Entero-Set and API 20E results were compared with those of the conventional media system, the Entero-Set performed as well as the API 20E in overall identification. Comparison of common biochemical tests among the various systems showed that citrate, arabinose, adonitol, inositol, and malonate gave correlations below 90%. The majority of the discrepancies were found among stock cultures. In addition, most discrepancies occurred with species of Enterobacter, Salmonella, Proteus, Klebsiella, and Serratia. Reproducibility studies showed the Entero-Set system to perform with a high degree of accuracy and reproducibility.

Bacteriological Techniques↗
PubMed1981

Evaluation of the repliscan system for Enterobacteriaceae identification.

A total of 1,013 isolates of Enterobacteriaceae were identified in parallel by the Repliscan (Cathra International, Ontario, Canada) and API 20E (Analytab Products, Plainview, N.Y.) systems. There was a 62% agreement at the genus level between the two systems. Of the 38% discrepant results, Repliscan classified 22% as "biochemical pattern not on file," 8% as a multiple-genus group which included the API 20E identification, and 8% as a genus other than that designated by API 20E. Relative to the various genera, Repliscan agreed with API 20E as follows: Escherichia coli, 80%; Klebsiella spp., 76%; Citrobacter spp., 75%; Proteus spp., 69%; Providencia spp., 54%; Serratia spp., 49%; Enterobacter spp., 25%; Shigella spp., 4%; and Salmonella spp., 0%. Repliscan identified 35% of Enterobacter spp. isolates as Citrobacter spp., 91% of Shigella spp. isolates as a multiple-choice-genus group, and 67% of Salmonella spp. isolates as "biochemical pattern not on file." Repliscan agreed with API 20E at the species level as follows: E. coli, 80%; Klebsiella spp., 56%; Citrobacter spp., 66%; Proteus spp., 55%; Providencia spp., 46%; Serratia spp., 39%; Enterobacter spp., 18%; Shigella spp., 4%; and Salmonella spp., 0%. These findings indicate that the Repliscan system in its present stage of development does not reliably identify the Enterobacteriaceae.

Bacteriological Techniques↗
PubMed1981

Evaluation of an automated, computerized system (automicrobic system) for Enterobacteriaceae identification.

The automated and computerized AutoMicrobic system (AMS; Vitek Systems, Inc., subsidiary of McDonnell Douglas, Hazelwood, Mo.) was evaluated as a means of identifying the Enterobacteriaceae. The Micro-ID system (General Diagnostics, Morris Plains, N.J.) and, when necessary, conventional tubed media were used for comparison. Identification by AMS and Micro-ID differed in only 12 of 1,528 isolates (0.8%). Disagreements occurred primarily with Enterobacter spp. Precision testing of the AMS showed only 1 of 72 tests (1.4%) deviating from the expected. The AMS was found to be an accurate and precise method for the identification of Enterobacteriaceae.

Autoanalysis↗
PubMed1981

Evaluation of the Repliscan II System for identification of Enterobacteriaceae.

In a precious report (Woolfrey et al., J Clin. Microbiol. 13:58-61, 1981), we indicated that the Repliscan system did not reliably identify Enterobacteriaceae. Recent improvements in the system prompted us to evaluate Repliscan II (Cathra International, Inc., St. Paul, Minn.) by using representative isolates of the population sample previously used to test the system. Isolates (692) representing eight genera were identified in parallel by the Repliscan II and API 20E (Analytab Products, Plainview, N.Y.) systems. Isolates given different identifications by the two systems were assigned reference identifications by using classical microbiological methods. Repliscan II identified 95.2% correctly, 4.0% incorrectly, and 0.7% as unknown. API 20E identified 99.4% correctly, 0.6% incorrectly, and none as unknown. Repliscan II correctly identified Salmonella and Shigella spp. to the genus level and isolates of six other genera to the species level as follows: Salmonella spp., 100%; Shigella spp., 97.7%; Escherichia spp., 95.2%; Citrobacter spp., 82.1%; Enterobacter spp., 85.2%; Klebsiella spp., 98.6%; Proteus spp., 97.2%; and Serratia spp., 97.9%. These findings indicate that Repliscan II is a significantly improved system and provides acceptable identification of Enterobacteriaceae.

Bacteriological Techniques↗
PubMed1981

Time-motion and cost comparison study of micro-ID, API 20E, and conventional biochemical testing in identification of Enterobacteriaceae.

A total of 730 Enterobacteriaceae strains isolated from 567 cultures were evaluated by a rapid kit method (Micro-ID; General Diagnostics, Morris Plains, N.J.; 4 h), an overnight incubation kit method (API 20E; Analytab Products, Plainview, N.Y.), and conventional biochemical test methodology (mostly overnight incubation and some rapid methods) to compare the amount of laboratory effort required, timing, and cost parameters. We assessed the amount of technologist time expended, the time sequence of culture reporting to physicians, the number of isolates requiring repeat testing or additional biochemical testing, the number of cultures held due to the need for identification of other organisms, the cost of total work-up, etc. Cultures evaluated included urines, respiratory cultures, wound cultures, body fluids, genital cultures, and cultures from miscellaneous categories. A total of 64% of the Enterobacteriaceae strains processed by the Micro-ID method could be identified within 24 h of receipt of the specimens in the clinical laboratories, in contrast to the need for an additional day required by the API or conventional biochemical methods. The Micro-ID method also required less technologist time (4.5 min) for set-up and interpretation than did either the API method (6 min) or conventional methods (7 min). Total direct costs (June 1981) per organism identified were: Micro-ID, $4.30; API 20E, $4.96; conventional biochemicals with commercially prepared media, $5.66. An estimate of 80% technologist time efficiency was made in all procedures.

Bacteriological Techniques↗
PubMed1982

Comparison of Enteric-Tek with API 20E and conventional methods for identification of Enterobacteriaceae.

Enteric-Tek (Flow Laboratories, Inc., Roslyn, N.Y.) is a new system designed for the identification of Enterobacteriaceae and consists of a round, multicompartmented plastic plate with 11 peripheral wells and 1 center well incorporating 14 biochemical reactions. Only one reagent (Kovacs reagent) is required to complete the biochemical test results for generation of a five-digit number for computer code identification. The identification accuracy of the Enteric-Tek system was compared with those obtained by conventional methods and API 20E. The Enteric-Tek system was found to be not only a convenient and simple method for rapid identification of Enterobacteriaceae, but also to be highly reliable, giving excellent identification performance as compared to API 20E and conventional methods.

Enterobacteriaceae↗
PubMed1982

Escherichia vulneris: a new species of Enterobacteriaceae associated with human wounds.

The name Escherichia vulneris sp. nov. (formerly called Alma group 1 and Enteric group 1 by the Centers for Disease Control and API group 2 by Analytab Products, Inc.) is proposed for a group of isolates from the United States and Canada, 74% of which were from human wounds. E. vulneris is a gram-negative, oxidase-negative, fermentative, motile rod with the characteristics of the family Enterobacteriaceae. Biochemical reactions characteristic of 61 E. vulneris strains were positive tests for methyl red, malonate, and lysine decarboxylase; a delayed positive test for arginine dihydrolase; acid production from d-mannitol, l-arabinose, raffinose, l-rhamnose, d-xylose, trehalose, cellobiose, and melibiose; negative tests for Voges-Proskauer, indole, urea, H(2)S, citrate, ornithine decarboxylase, phenylalanine deaminase, and DNase; and no acid from dulcitol, adonitol, myo-inositol, and d-sorbitol. Two-thirds of the strains produced yellow pigment. Most strains gave negative or delayed positive reactions in tests for lactose, sucrose, and KCN. The E. vulneris strains tested were resistant to penicillin and clindamycin, were resistant or showed intermediate zones of inhibition to carbenicillin and erythromycin, and were susceptible to 14 other antibiotics. DNA relatedness of 15 E. vulneris strains to the type strain averaged 75% in reactions at 60 degrees C and 69% in reactions at 75 degrees C, indicating that they comprise a separate species. DNA relatedness to other species in the family Enterobacteriaceae was 6 to 39%, an indication that this new species belongs in the family. E. vulneris showed the highest relatedness to species of Escherichia (25 to 39%) and Enterobacter (24 to 35%). On the basis of biochemical similarity, the new species was placed in the genus Escherichia. The type strain of E. vulneris is ATCC 33821 (CDC 875-72).

Anti-Bacterial Agents↗
PubMed1982

Reliability of early identifications obtained with Enterobacteriaceae-plus biochemical cards in the automicrobic system.

The AutoMicrobic system (AMS) is capable of identifying most Enterobacteriaceae within 8 h and many glucose-nonfermenting, gram-negative bacilli after 13 h of incubation. Early preliminary results can be readily obtained from the computer as the tests incubate. Data with 1,023 bacterial isolates were reviewed to determine the relative accuracy of 4-, 6-, 8-, 10-, and 13-h identifications. All AutoMicrobic system identifications with probability (P) values of less than 0.80 were considered equivocal responses which needed supplementary tests before a final report could be issued. Analysis of our data suggests that early identifications of Morganella morganii, Acinetobacter sp., Yersinia spp., Salmonella spp. (other than Salmonella typhi), Shigella spp. (other than Shigella sonnei), Enterobacter agglomerans, Pseudomonas spp. (other than Pseudomonas aeruginosa or Pseudomonas maltophilia), Klebsiella spp. (other than Klebsiella pneumoniae or Klebsiella oxytoca), Citrobacter amalonauticus, Serratia liquefaciens, or Vibrio spp. Should be considered nonspecific responses, even when P greater than or equal to 0.80. Other identifications reported after 4 h were 96% accurate. At least half of our isolates (60% of our Enterobacteriaceae) could be identified reliably within 4 h, the remaining isolates required longer incubation.

Bacteria↗
PubMed1983

Evaluation of the AutoSCAN-3 and Sceptor systems for Enterobacteriaceae identification.

To evaluate the accuracy and cost effectiveness of the AutoSCAN-3 (Micro-Scan Systems of America, Sacramento, Calif.) and Sceptor (BBL Microbiology Systems, Cockeysville, Md.) systems for identification of members of the Enterobacteriaceae, we performed parallel tests on 678 stock cultures of well-characterized clinical isolates of Enterobacteriaceae. Automated results by AutoSCAN-3 correctly identified 95.1% at the genus level and 94.9% at the species level. However, 15 of 42 Shigella isolates were misidentified as members of other genera. In contrast to the automated results, visual interpretation of panels produced 97.9% agreement at the genus level, missing only three Shigella isolates. Sceptor correctly identified 96.8% at the genus level and 93.4% at the species level. Of 42 Shigella isolates, 3 were missed and were designated as Salmonella spp. Although all Salmonella spp. were correctly identified, six other isolates were misidentified as Salmonella spp. Test costs were found to be comparable for each system, with the cost per test increasing markedly with fewer than 10 to 15 tests performed per day.

Bacteriological Techniques↗
PubMed1984

Moellerella wisconsensis, a new genus and species of Enterobacteriaceae found in human stool specimens.

The name Moellerella wisconsensis is proposed for a group of the family Enterobacteriaceae previously called enteric group 46. The species name, wisconsensis, was coined because six of the nine strains were isolated in Wisconsin. M. wisconsensis strains were negative for indole production, Voges-Proskauer, H2S production, urea, phenylalanine deaminase, lysine and ornithine decarboxylases, arginine dihydrolase, gas production from D-glucose, acid production from trehalose, and motility; the strains were positive for methyl red, citrate (Simmons), and acid production from lactose and raffinose and resistant to colistin. DNAs from five strains of M. wisconsensis were highly related (80 to 93% in reactions assayed on hydroxyapatite at 60 degrees C and 78 to 97% at 75 degrees C) to 32P-labeled DNA of the proposed type strain (CDC 2896-78, ATCC 35017). Labeled DNA from this type strain was only 2 to 32% related (at 60 degrees C) to DNA from 49 strains of named and unnamed species of Enterobacteriaceae. Eight of nine M. wisconsensis strains were isolated from human stool samples. Clinical information on one strain was available, and it was found to be associated with a case of diarrhea. On MacConkey agar, colonies of M. wisconsensis were bright red with precipitated bile around them and thus were indistinguishable from Escherichia coli colonies. Future studies should focus on the isolation of this new organism and its relationship to human disease.

Adult↗
PubMed1975

Motility-indole-lysine medium for presumptive identification of enteric pathogens of Enterobacteriaceae.

Detection of lysine decarboxylase activity is a useful supplement to reactions on triple sugar-iron (TSI) and urea agars in the initial examination of suspected pathogenic isolates from fecal cultures. Owing to the added value of motility and indole production in the differentiation of enteric pathogens, we prepared and evaluated a motility-indole-lysine (MIL) medium. The following 890 organisms were tested: 264 Shigella, 2 Edwardsiella, 182 Salmonella enteritidis, 235 S. typhi, 3 Arizona, 32 Yersinia enterocolitica, and 172 other members of the family Enterobacteriaceae. With few exceptions the MIL medium gave the same results as the standard motility, indole, and lysine decarboxylase (Moeller) test media. All discrepancies were with the indole reaction, which was weak in 2 of 67 strains of Escherichia coli and falsely negative in 6 of 32 strains of Y. enterocolitica. When both TSI agar and lysine-iron agar (LIA) slants are used in the evaluation isolates from fecal cultures, detection of H2S is duplicated. Both LIA and MIL medium detect lysine decarboxylase and deaminase activity equally well. Because of its ability to detect motility and indole production, the MIL medium is more useful than LIA when used with TSI agar. The combination of TSI agar, MIL medium, and urea agar enables reliable initial recognition of enteric pathogens of the Enterobacteriaceae.

Carboxy-Lyases↗
PubMed1984

Comparative evaluation of the Eiken and API 20E systems and conventional methods for identification of members of the family Enterobacteriaceae.

To evaluate the accuracy and utility of the Eiken Systek No. 1 (Eiken system; Eiken Chemical Co., Ltd., Tokyo, Japan), we conducted a clinical comparison, with 345 Enterobacteriaceae isolates, of the Eiken System with API 20E (Analytab Products, Inc., Plainview, N.Y.) and conventional methods. The Eiken system is a 21-biochemical-test battery tray stored at 25 degrees C and inoculated in one step. It is similar to the API 20E except that the Eiken system contains malonate, adonitol, and maltose; lacks gelatin, sucrose, melibiose, amygdalin, and arabinose; and uses reagent strips instead of liquid reagents. The API 20E and Eiken systems correctly identified 339 (97.7%) and 276 (79.5%), respectively, and misidentified 3 (0.9%) and 13 (3.7%), respectively, of the isolates. There were no identification codes for 5 (1.4%) organisms with the API 20E and 58 (16.7%) organisms with the Eiken system; of these latter unidentified organisms, 42 were identified as Proteus spp., Morganella sp., and Providencia rettgeri by conventional methods. There was no significant difference between the two rapid systems in total time required for inoculation and reading. Modifications for interpretation of decarboxylase and oxidase tests were needed for the Eiken system, and manipulation of reagent strips required considerable dexterity. However, the Eiken system was easier to inoculate than the API 20E, and, with minor increases in the data base to include more of the Proteus and Morganella spp. and P. rettgeri, the system should be reliable for identification of members of the family Enterobacteriaceae.

Computers↗