SummarySepsis, a life‐threatening systemic infection, requires quick treatment. Gram‐negative bacteria (GNB) are the major causative pathogens and their endotoxin can be a surrogate biomarker for diagnosis. We explored a fast identification of GNB by first culturing blood to increase endotoxin levels and then detecting endotoxin by Tachypleus amebocyte lysate (TAL) with kinetic turbidimetric assay (KT‐TAL). Heating samples could significantly increase the endotoxin released from GNB; speed and time of centrifugation, and sample dilution could affect the endotoxin results. At a high GNB load, endotoxin was detected 3 h after culture, 6.5 h earlier than the BD BACTEC blood culture system detecting GNB. At a low GNB load, endotoxin was detected at 9 h after culture, 13 h earlier than by the BD BACTEC system. In a sepsis patient with Acinetobacter baumannii, we detected endotoxin at 12 h after culture, while the BD BACTEC system needed 28.5 h for detection, allowing physicians an earlier decision on appropriate treatment.
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