ABSTRACT
Objectives: As time to appropriate antimicrobial therapy is major to reduce sepsis mortality, there is great interest in the development of tools for direct identification (ID) and antimicrobial susceptibility testing (AST) of positive blood cultures (PBC). Very recently, the FAST™ System (Qvella) has been developed to isolate and concentrate microorganisms directly from PBCs, resulting in the recovery of a Liquid Colony™ (LC) within 30 min. The LC can be used as equivalent of an overnight subcultured colony for downstream testing. We aimed to evaluate the performances of the FAST™ System and FAST-PBC Prep™ cartridges by testing the resulting LC for direct ID, AST and rapid resistance detection. Materials and methods: Prospectively, FAST™ System testing was carried out on each patient's first PBC with a monomicrobial Gram-stain result. In the second arm of the study, FAST™ System testing was carried out on blood cultures spiked with multidrug-resistant bacteria. Downstream testing using the LC included MALDI-TOF MS ID with the Bruker Biotyper® smart system, rapid resistance detection testing including the Abbott Diagnostics Clearview™ PBP2a SA Culture Colony Test (PBP2a) and the Bio-Rad ßLACTA™ Test (ßLT). AST was performed using the Becton Dickinson Phoenix™ System or by Bio-Rad disk diffusion using filter paper disk following EUCAST 2020 breakpoint criteria. Results: FAST™ System testing was completed on 198 prospective PBCs and 80 spiked blood cultures. After exclusion of polymicrobial blood cultures, performance evaluation compared with standard of care results was carried out on 266 PBCs. Concordant, erroneous and no ID results included 238/266 (89.5%), 1/266 (0.4%), 27/266 (10.2%) PBCs, respectively. Sensitivity and specificity for PBP2a were 100% (10/10) and 75% (15/20), respectively. Sensitivity and specificity for ßLT were 95.8% (23/24) and 100% (42/42), respectively. Categorical agreement for all 160 tested strains was 98% (2299/2346) with 1.2% (8/657) very major errors and 0.7% (10/1347) major errors. Conclusion: FAST™ System testing is a reliable approach for direct downstream testing of PBCs including MALDI-TOF MS ID, BD Phoenix™ and Bio-Rad disk diffusion AST as well as rapid resistance testing assays. Next steps include optimal integration of the FAST™ System in the PBC workflow with a view toward clinical studies.
ABSTRACT
PURPOSE: Pulmonary infections due to non-tuberculous mycobacteria (NTM) are an emerging issue in the cystic fibrosis (CF) population. Due to bacterial and fungal overgrowth, isolation of mycobacteria from the sputum samples of these patients remains challenging. RGM medium, a novel agar-based culture medium was evaluated for the isolation of NTM from sputum samples of CF patients. METHODOLOGY: Sputum samples were inoculated onto RGM medium and conventional Mycobacterial Growth Indicator Tube (MGIT™, Becton Dickinson, USA). Agar plates were incubated at 35⯰C and growth was recorded once a week during 42 days. We compared the yield of the two media. RESULTS: 217 samples were obtained from 124 CF patients. 20 samples (13 patients) had a positive culture for NTM. 79/217 (36.4%) MGIT had to be discontinued due to contamination compared to 18/217 (8.3%) for RGM. We reported equivalent NTM detection performances for RGM and MGIT (Pâ¯=â¯0.579): these media enabled the isolation of 15 and 12 NTM strains respectively. CONCLUSION: RGM medium increases the proportion of interpretable results and the number of NTM cultured. Taking into account the non-inferiority compared to conventional methods and ease of use of RGM medium, we estimate that this test can replace current approaches for the screening of NTM among people with CF. Additionally, RGM provides semi-quantitative results (number of colonies) and information on the morphology of colonies, which may be clinically relevant information.
ABSTRACT
A rapid preparation procedure was validated for MALDI-TOF MS identification followed by an antimicrobial susceptibility testing directly from a non-selective enrichment broth of sterile fluid samples associated with negative classical cultures. This method can be easily integrated in the laboratory routine allowing precious time gain for microbe identification and subsequent adequate treatment initiation.
