IntroductionInfections caused by multidrug-resistant microorganisms have become increasingly common in hospital environments around the world. Gram-negative bacilli stands out among multidrug-resistant bacteria mostly due to the production of carbapenemase enzymes which lead to resistance to most β-lactam antibiotics including the carbapenems. As a consequence, polymyxins have been reintroduced in the clinic as a last resort to treat infections caused by Gram-negative bacilli resistant to carbapenems. However, the only reliable method to evaluate the susceptibility to polymyxins is the broth microdilution, a laborious and time-consuming technique. Among infections caused by multidrug-resistant bacteria, bloodstream infections are the most worrisome as they can lead to sepsis and septic shock with high mortality rates.ObjectiveConsidering the severity of sepsis and the need for a treatment guided for the susceptibility test in vitro, this work aimed to evaluate a rapid method of polymyxins susceptibility either from colonies grown on agar or directly from positive blood culture bottles using the technology of MALDI-TOF.MethodsThe method was based on the “direct on target microdroplets growth assay” (DOT-MGA) originally developed by Idelevich and collaborators with some modifications (Adapted DOT-MGA). Isolates of Enterobacterales and non-fermenting Gram-negative bacilli resistant to carbapenems were obtained from patients attending a tertiary care hospital in southern Brazil and tested as follows: 122 isolates from colonies grown on agar plates and 117 isolates directly from spiked positive blood cultures.ResultsThe adapted DOT-MGA presented 95 and 100% of categorical agreement considering the colonies grown on agar plates and directly from positive blood cultures, respectively.DiscussionThe adapted DOT-MGA test proved to be a reliable technique to evaluate the susceptibility to polymyxins to be used in microbiology laboratories with the MALDI-TOF equipment.
Determination of the susceptibility profile of isolates of Candida from blood culture bottles is extremely important for correctly guiding patient pharmacotherapy. The aim of this study was to compare the results of analysis of Candida isolated directly from blood culture bottles by the VITEK MS MALDI-TOF identification system and the fluconazole disk diffusion assay with those of standard identification methods. Testing directly from the bottle allowed results 24 to 48 hours quicker than the standard method. There was a categorical agreement of 51.64% (47 of 91 samples) between the results of analysis directly from the bottle and analysis by the standard method. Regarding species identification, there was 96.15% agreement for Candida parapsilosis (25 of 26 samples). Categorical agreement between the rapid and standard disk diffusion methods was 95%, and the agreement between the rapid disk diffusion method and the broth microdilution method was 97%. Only minor errors in the rapid method were observed: 3 (5%) in the standard disk diffusion method and 2 (3%) in the broth microdilution method. Our study concluded that the rapid disk diffusion method for fluconazole is a fast, easy, reproducible, and consistent method. Its timely implementation for testing antifungal agents in the clinical microbiology laboratory can help reduce profile release times, thus helping to determine the most appropriate antifungal treatment.
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