A new chromogenic endotoxin-specific assay using recombined limulus coagulation enzymes and its clinical applications

Clin Chim Acta. 1985 Jun 30;149(1):55-65. doi: 10.1016/0009-8981(85)90273-6.

Abstract

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.

MeSH terms

  • Arthropod Proteins
  • Chromogenic Compounds
  • Endopeptidases / isolation & purification
  • Endotoxins / analysis*
  • Endotoxins / poisoning
  • Escherichia coli
  • Horseshoe Crabs / enzymology
  • Humans
  • Lectins / isolation & purification
  • Limulus Test*
  • Shock, Septic / blood

Substances

  • Arthropod Proteins
  • Chromogenic Compounds
  • Endotoxins
  • Lectins
  • Endopeptidases