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Healthcare systems spend considerable resources collecting and processing blood cultures for the detection of blood stream pathogens. The process is initiated with the collection of blood cultures that depend upon proper skin disinfection, collection of an adequate number of specimens and volume of blood, and prompt processing in a sensitive culture system. Complementing blood cultures and gaining in use are techniques such as nucleic acid amplification tests and mass spectroscopy that allow clinical laboratories to detect and identify organisms from blood cultures substantially faster than conventional systems. Furthermore, certain resistance mutations can be detected within hours of organism detection, thus providing valuable guidance to clinicians who strive to initiate the appropriate antimicrobial therapy as rapidly as possible, and who wish to discontinue unnecessary drugs expeditiously. Molecular and mass spectroscopy techniques are changing sepsis diagnosis rapidly and will provide far more specific information far more quickly, but the performance characteristics of these systems must be understood by intensivists who use such information to guide their patient management.