A novel approach for fast detection of sepsis with Gram-negative bacterial infection.

Sepsis, 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.