Emerging yeast pathogens are favoured by increasing numbers of immunocompromised patients and by certain current medical practices. These yeasts differ in their antifungal drug susceptibilities, and rapid species identification is imperative. A large variety of methods have been developed with the aim of facilitating rapid, accurate yeast identification. Significant recent commercial introductions have included species-specific direct enzymatic colour tests, differential chromogenic isolation plates, direct immunological tests, and enhanced manual and automated biochemical and enzymatic panels. Chromogenic isolation media demonstrate better detection rates of yeasts in mixed cultures than traditional media, and allow the direct identification of Candida albicans by means of colony colour. Comparative evaluation of rapid methods for C. albicans identification, including the germ tube test, shows that chromogenic media may be economically advantageous. Accurate tests for single species include the Bichrolatex Albicans and Krusei Color tests, both immunologically based, as well as the Remel Rapid Trehalose Assimilation Broth for C. glabrata. Among broad-spectrum tests, the RapID Yeast Plus system gives same-day identification of clinical yeasts, but performance depends on inoculum density and geographic isolate source. The API 20 C AUX system is considered a reference method, but newer systems such as Auxacolor and Fungichrom are as accurate and are more convenient. Among automated systems, the ID 32 C strip, the Vitek Yeast Biochemical Card and the Vitek 2 ID-YST system correctly identify >93% of common yeasts, but the ID-YST is the most accurate with uncommon yeasts, including C. dubliniensis. Spectroscopic methods such as Fourier transformed-infrared spectroscopy offer potential advantages for the future. Overall, the advantages of rapid yeast identification methods include relative simplicity and low cost. For all rapid methods, meticulous, standardized multicenter comparisons are needed before tests are fully accepted.

In order to explore pathologies possibly associated with vitamin K deficiency, several monoclonal antibodies (mAbs) were produced against human Desgamma-Carboxy-Prothrombin (DCP). One of these mAbs, designated C4B6, detected DCP forms in the presence of Calcium ions, confirmed by comparison with the patterns of two electrophoretic techniques: Affino-Immuno-Electrophoresis (CAIE) and Polyacrylamide Gel Electrophoresis followed by Electro-blotting (PAGE-Blot). An Enzyme-Linked-Imunosorbent Assay (ELISA) using mAb C4B6 has been developed, optimized and standardized. It has proven to be specific for DCP forms and has a minimum sensitivity of 0.156 A.U/ml.

Five yeast strains, causally associated with septicemia and death in a patient after peritonitis, were identified as Candida lusitaniae van Uden et do Carmo-Sousa by standard methods. The organism was initially susceptible to 5-fluorocytosine but strongly resistant to amphotericin B, requiring 50 micrograms/ml for complete inhibition at 48 h.

Six commercially available systems for the identification of yeasts were evaluated using 133 clinical isolates and four reference strains that had been previously identified by conventional methods and 19 recent clinical isolates that had been identified by the ID32C system (bioMérieux, France). The total identification rates (TIR) established for the total number of strains tested and the database identification rates (DBIR) established for the strains included in the respective manufacturer databases were both determined. After incubation for 4 h, the TIR and DBIR were 78% and 84%, respectively, for the RapID Yeast Plus system (Innovative Diagnostic Systems, USA). After incubation for 24 h, the TIR and DBIR were 32% and 32%, respectively, for the ID32C, 65% and 67% for the Auxacolor system (Sanofi Diagnostics Pasteur, France), 62% and 65% for the Fungichrom I system (International Microbio, France), 52% and 65% for the Fungifast I twin system (International Microbio), and 62% and 68% for the API Candida system (bioMérieux). The maximum TIR and DBIR (+/- 1%) obtained after incubation for 48 h were 86% and 88% for the Auxacolor, 85% and 89% for the Fungichrom I, 78% and 98% for the Fungifast I twin, and 82% and 91% for the API Candida. For the ID32C, the maximum TIR and DBIR were 98% and 98%, respectively, but these values were obtained only after 72 h of incubation. In addition, the six systems varied in their ease of use and readings. In conclusion, based on results obtained with 156 strains, the Auxacolor and Fungichrom systems seem the most appropriate for use in a clinical microbiology laboratory, due to their ease of use and reading, their rapidity, their cost per test, and their relatively high TIR results, which indicated acceptable performance with strains frequently isolated in our hospital. For a reference identification, the ID32C remains the sole system usable.

No abstract

The new micromethod for yeast susceptibility testing, MYCOTOTAL, was evaluated with 10 reference strains in seven laboratories. Ready-to-use microtitration plates and the same synthetic medium were used with two dilutions of imidazoles, flucytosine, and amphotericin B, permitting the categorization of each strain as susceptible, intermediate, or resistant. The results were compared with the MIC for each reference strain, and the repeatability and reproducibility were evaluated. The yeasts tested presenting different patterns of susceptibilities in reference MICs included six strains of Candida albicans, two strains of Candida tropicalis, one strain of Candida parapsiosis, and one strain of Torulopsis glabrata. For 4,200 antifungal agent-yeast results, the repeatability was 99.3% and the reproducibility was 96.3%. The correlation between the reference MICs and the category results was 91.5% for seven laboratories (and 92.7% for six laboratories excluding the laboratory which did not follow exactly the same protocol). We observed only 7.9% minor discrepancies, 0.5% (0.29% for six laboratories) major discrepancies, and 0.1% uninterpretable results. The percentages of concording results were similar for each strain and each antifungal agent tested. The overall results indicated that MYCOTOTAL was a reliable and reproducible method, well correlated with reference MICs. This ready-to-use micromethod with the same medium for all antifungal agents would be an important step in the necessary standardization of yeast susceptibility testing.

Six commercially available systems for the identification of yeasts were evaluated using 133 clinical isolates and four reference strains that had been previously identified by conventional methods and 19 recent clinical isolates that had been identified by the ID32C system (bioMérieux, France). The total identification rates (TIR) established for the total number of strains tested and the database identification rates (DBIR) established for the strains included in the respective manufacturer databases were both determined. After incubation for 4 h, the TIR and DBIR were 78% and 84%, respectively, for the RapID Yeast Plus system (Innovative Diagnostic Systems, USA). After incubation for 24 h, the TIR and DBIR were 32% and 32%, respectively, for the ID32C, 65% and 67% for the Auxacolor system (Sanofi Diagnostics Pasteur, France), 62% and 65% for the Fungichrom I system (International Microbio, France), 52% and 65% for the Fungifast I twin system (International Microbio), and 62% and 68% for the API Candida system (bioMérieux). The maximum TIR and DBIR (+/- 1%) obtained after incubation for 48 h were 86% and 88% for the Auxacolor, 85% and 89% for the Fungichrom I, 78% and 98% for the Fungifast I twin, and 82% and 91% for the API Candida. For the ID32C, the maximum TIR and DBIR were 98% and 98%, respectively, but these values were obtained only after 72 h of incubation. In addition, the six systems varied in their ease of use and readings. In conclusion, based on results obtained with 156 strains, the Auxacolor and Fungichrom systems seem the most appropriate for use in a clinical microbiology laboratory, due to their ease of use and reading, their rapidity, their cost per test, and their relatively high TIR results, which indicated acceptable performance with strains frequently isolated in our hospital. For a reference identification, the ID32C remains the sole system usable.

In this work we were primarily concerned with antigens of Bacteroides strains belonging to the B. fragilis group. The following two antisera were produced: anti-Bacteroides fragilis ElT (T = type strain) and anti-Bacteroides distasonis. Reference patterns for both of these species were established by crossed immunoelectrophoresis, and these patterns yielded 62 and 70 precipitates, respectively. The crossed reactions of B. fragilis ElT and B . distasonis with Bacteroides fragilis E2, Bacteroides thetaiotaomicron, Bacteroides ovatus and Bacteroides vulgatus were studied to test both for total antigens and for certain enzymes (esterase, phosphatase, malate dehydrogenase, and glucose-6-phosphate dehydrogenase). Our results showed that B. thetaiotaomicron is the species which is closest to B. fragilis, demonstrating more than 70% common antigens, whereas none of the species examined was very closely related to B. distasonis. Species-specific antigens, such as the phosphatase of B. fragilis, as well as antigens common to the group, such as malate dehydrogenase, were also demonstrated in crossed immunoelectrophoresis. This may allow immunological identification of each species.Anaerobic bacteria are responsible for at least 10% of all bacterial infections, and Bacteroides species alone represent 27% of the anaerobes isolated from pathological materials (22). Bacteroides strains belonging to the B. fragilis group are most often responsible for intestinal infections (6, 27). The most frequently isolated species is Bacteroides fragilis, although this species is a minor component of the fecal flora, in which Bacteroides vulgatus and Bacteroides thetaiotaomicron are the dominant species (18). This observation has led some authors to seek virulence factors in B . fragilis (notably in the capsule) that are lacking in the other species of the group (13, 19).Classification of the B . fragilis group has been modified often. The numerous phenotypic properties which members of this group have in common account for the proposal of subspecies of B . fragilis (17). However, deoxyribonucleic acid-deoxyribonucleic acid homology studies showed a diversity (3) which led to reclassification of B . fragilis, Bacteroides ovatus, B. vulgatus, B . thetaiotaomicron, and Bacteroides distasonis as distinct species (12).In 1971, Beerens et al. (1) proposed a serological classification for the B. fragilis group, which was based on agglutination or gel double-diffusion tests in the presence of a specific antiserum. Lambe and Moroz (15) tested for the presence of thermostable antigens that permitted classification of all B . fragilis strains without cross-reactions with B . distasonis, B . ovatus, and B . thetaiotaomicron. Other authors have studied the lipopolysaccharide fractions or outer membrane complex of B. fragilis by using either sodium dodecyl sulfate-polyacrylamide gel electrophoresis (4, 14, 21) or immunochemical techniques (5, 10).All of these studies demonstrated the heterogeneity which exists within the B . fragilis group, as well as t...