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Preparation of a blood culture pellet for rapid bacterial identification and antibiotic susceptibility testing.
Journal of Visualized Experiments : Jove
Publication types: Journal Article ; Video-Audio Media Publication Status: epublish Video de protocol expérimental dans Jove
Bloodstream infections and sepsis are a major cause of morbidity and mortality. The successful outcome of patients suffering from bacteremia depends on a rapid identification of the infectious agent to guide optimal antibiotic treatment. The analysis of Gram stains from positive blood culture can be rapidly conducted and already significantly impact the antibiotic regimen. However, the accurate identification of the infectious agent is still required to establish the optimal targeted treatment. We present here a simple and fast bacterial pellet preparation from a positive blood culture that can be used as a sample for several essential downstream applications such as identification by MALDI-TOF MS, antibiotic susceptibility testing (AST) by disc diffusion assay or automated AST systems and by automated PCR-based diagnostic testing. The performance of these different identification and AST systems applied directly on the blood culture bacterial pellets is very similar to the performance normally obtained from isolated colonies grown on agar plates. Compared to conventional approaches, the rapid acquisition of a bacterial pellet significantly reduces the time to report both identification and AST. Thus, following blood culture positivity, identification by MALDI-TOF can be reported within less than 1 hr whereas results of AST by automated AST systems or disc diffusion assays within 8 to 18 hr, respectively. Similarly, the results of a rapid PCR-based assay can be communicated to the clinicians less than 2 hr following the report of a bacteremia. Together, these results demonstrate that the rapid preparation of a blood culture bacterial pellet has a significant impact on the identification and AST turnaround time and thus on the successful outcome of patients suffering from bloodstream infections.
Anti-Bacterial Agents/pharmacology, Bacteremia/microbiology, Bacteriological Techniques/methods, Blood/microbiology, Gram-Negative Bacterial Infections/blood, Gram-Negative Bacterial Infections/microbiology, Gram-Positive Bacterial Infections/blood, Gram-Positive Bacterial Infections/microbiology, Humans, Microbial Sensitivity Tests/methods, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods
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