A new chromogenic endotoxin-specific assay using recombined limulus coagulation enzymes and its clinical applications.
A conventional limulus test is not specific to endotoxin because of the presence in amebocyte lysate of a (1----3)-beta-D-glucan-sensitive factor. By fractionating coagulation enzymes in the lysate and recombining only those factors involved in endotoxin-induced coagulation, we have developed a new test specific to endotoxin. The recombined enzymes reacted only with endotoxin, and not with fungal polysaccharides. Conventional amebocyte lysate, on the other hand, reacted with both of them. A good linearity was obtained with this method between endotoxin concentration and absorbance with a sensitivity of 1 pg/ml of Escherichia coli 0111:B4 endotoxin. The regression lines for different types of endotoxins were parallel to one another. For the correct diagnosis of endotoxemia, this new test has a definite advantage over the one using whole amebocyte lysate.
D060829:Arthropod Proteins; D002863:Chromogenic Compounds; D010450:Endopeptidases / Q000302:isolation & purification; D004731:Endotoxins / Q000032:analysis* / Q000506:poisoning; D004926:Escherichia coli; D006737:Horseshoe Crabs / Q000201:enzymology; D006801:Humans; D037102:Lectins / Q000302:isolation & purification; D008033:Limulus Test; D012772:Shock, Septic / Q000097:blood
D060829:Arthropod Proteins; D002863:Chromogenic Compounds; D004731:Endotoxins; D037102:Lectins; D010450:Endopeptidases