ABSTRACT
Accuracy of the Vitek 2 automated system (bioMérieux Vitek, USA) for rapid identification of bacteria was evaluated using a collection of 858 epidemiologically unrelated gram-negative and 99 gram-positive clinical isolates. Isolates were tested after subculturing to ensure purity. Conventional agar-based biochemical tests (Steers replicator) were used as a reference method of identification. Gram-negative bacteria were identified to the species level with 95.3% accuracy by the system ( Enterobacteriaceae, 95.9%; and non- Enterobacteriaceae, 92.5%), and gram-positive isolates with 72% accuracy. Although Vitek 2 identified routine clinical isolates of gram-negative bacilli and Enterococcus faecalis and Enterococcus faecium reliably, rapidly, and reproducibly, improvement is required in the identification of less common species of enterococci and viridans group streptococci.
Subject(s)
Bacterial Typing Techniques/methods , Bacterial Typing Techniques/standards , Streptococcaceae/classification , Streptococcaceae/isolation & purification , Automation , Enterobacteriaceae/classification , Enterobacteriaceae/isolation & purification , Humans , Quality Control , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
With the recent dramatic rise in fluconazole use, there has been an increase in Candida species resistant to that agent. This has led to the clinical development of newer triazoles such as voriconazole that have greater potency and a broader spectrum of activity. We therefore hypothesized that fluconazole-resistant Candida albicans and Candida krusei would be susceptible to voriconazole. Susceptibility testing was performed on 205 isolates of C. albicans collected from 1984 to 1995, and on C. albicans and C. krusei that were identified as fluconazole resistant since 1995. The anti fungal agents used were amphotericin B, 5-flucytosine, itraconazole, ketoconazole, fluconazole and voriconazole. Three C. albicans and 26 C. krusei isolates had a minimum inhibitory concentration (MIC) >/=20 mg/l and were defined as fluconazole resistant. Of these, 28 isolates were susceptible to =2.5 mg/l voriconazole, with a mean MIC of 0.78 mg/l. The mean amphotericin B MIC for these same strains was 0.98 mg/l. Only one isolate of C. krusei was relatively resistant to voriconazole with a MIC of 5 mg/l. Of these 29 isolates, there were ten amphotericin-resistant strains of C. krusei (MIC>2 mg/l) but all were susceptible to =2.5 mg/l voriconazole. The higher potency of voriconazole may be useful in the treatment of fluconazole- and amphotericin-resistant C. albicans and C. krusei.
Subject(s)
Antifungal Agents/pharmacology , Candida albicans/drug effects , Pyrimidines/pharmacology , Triazoles/pharmacology , Candida albicans/isolation & purification , Drug Resistance, Microbial , Fluconazole/pharmacology , Humans , Microbial Sensitivity Tests , VoriconazoleABSTRACT
Drug resistance is emerging in many important microbial pathogens, including Candida albicans. We performed fungal susceptibility tests with archived isolates obtained from 1984 through 1993 and fresh clinical isolates obtained from 1994 through 1997 by testing their susceptibilities to fluconazole, ketoconazole, and miconazole and compared the results to the rate of fluconazole use. All isolates recovered prior to 1993 were susceptible to fluconazole. Within 3 years of widespread azole use, we detected resistance to all agents in this class. In order to assess the current prevalence of resistant isolates in our hematologic malignancy and transplant patients, we obtained rectal swabs from hospitalized, non-AIDS, immunocompromised patients between June 1995 and January 1996. The swabs were inoculated onto sheep's blood agar plates containing 10 microg of vancomycin and 20 microg of gentamicin/ml of agar. One hundred one yeasts were recovered from 97 patients and were tested for their susceptibilities to amphotericin B, fluconazole, flucytosine, ketoconazole, and miconazole. The susceptibility pattern was then compared to those for all clinical isolates obtained throughout the medical center. The antifungal drug histories for each patient were also assessed. The yeasts from this surveillance study were at least as susceptible as the overall hospital strains. There did not appear to be a direct linkage between prior receipt of antifungal agent therapy and carriage of a new, drug-resistant isolate. Increased resistance to newer antifungal agents has occurred at our medical center, but it is not focal to any high-risk patient population that we studied. Monitoring of susceptibility to antifungal agents appears to be necessary for optimizing clinical therapeutic decision making.
Subject(s)
Antifungal Agents/therapeutic use , Azoles/therapeutic use , Mycoses/drug therapy , Mycoses/microbiology , Neoplasms/complications , Yeasts/drug effects , Candida/drug effects , Candida albicans/drug effects , Drug Resistance, Microbial , Fluconazole/adverse effects , Fluconazole/therapeutic use , Humans , Imidazoles/therapeutic use , Microbial Sensitivity TestsABSTRACT
Nine selected isolates of Candida albicans were tested for their susceptibilities to amphotericin B and fluconazole by using three methods to assess the effect of incubation time and buffer concentration. By using a microdilution method with 0.0165 M 3-(N-morpholino)propanesulfonic acid (MOPS) and a 24-h incubation time, all of the isolates were found to be susceptible to amphotericin B and fluconazole. After 48 h of incubation, all isolates were still susceptible to amphotericin B. Seven of the nine isolates were resistant to fluconazole, and for the remaining two isolates, MICs increased by fourfold or more but the isolates remained susceptible (MIC, < or = 10 microg/ml). The nine isolates, along with three control strains, were further tested against amphotericin B and fluconazole by a standard broth macrodilution method with both 0.165 and 0.0165 M MOPS. The susceptibility results for fluconazole by the broth macrodilution method with the lower MOPS concentration correlated with the results of the 24-h broth microdilution method for determination of susceptibility or resistance in eight of nine tests and with the results of the 48 h broth microdilution method in three of nine tests. The results of the broth macrodilution method with the standard MOPS concentration did not correlate with any of the results obtained by the 24-h broth microdilution but correlated with results of seven of nine tests by the 48-h broth microdilution method. All nine test strains appeared to be susceptible when they were examined by a flow cytometric method. For clinical yeast susceptibility testing in microdilution panels, the 0.0165 M MOPS concentration combined with 24 h of incubation appeared to be the method of choice. The lower MOPS concentration may also be a useful modification to the tentative broth macrodilution method of the National Committee for Clinical Laboratory Standards. Use of the higher buffer concentration or longer incubation time may lead to false in vitro resistance for agents like fluconazole.