[Development of an endotoxin-specific Limulus amebocyte lysate test blocking beta-glucan-mediated pathway by carboxymethylated curdlan and its application].
We developed a simple new endotoxin-specific assay method that uses Limulus amebocyte lysate (LAL) containing a sufficient amount of a water-soluble (1----3)-beta-D-glucan derivative as a blocker of the (1----3)-beta-D-glucan-mediated coagulation pathway. The addition of 0.1 mg/ml or more of carboxymethylated (1----3)-beta-D-glucan completely blocked the activation of LAL by (1----3)-beta-D-glucan itself. The assay of endotoxin was unaffected by the presence of 1 mg/ml carboxymethylated (1----3)-beta-D-glucan. Spiked endotoxin was recovered well from beta-glucans by the turbidimetric kinetic method with LAL containing 1 mg/ml of carboxymethylated (1----3)-beta-D-glucan. Besides, this new LAL formulation was applied for an endotoxin-specific assay by the conventional gel-clot method or the chromogenic method. Gram-negative bacteria were specifically detected by the turbidimetric kinetic method with the LAL formulation. This LAL formulation may be used for an endotoxin-specific assay not only in pharmacology but also in clinical microbiology.
Tsuchiya
M
M
Osaka Research Laboratory, Wako Pure Chemical Industries, Ltd., Hyogo, Japan.
Takaoka
A
A
Tokioka
N
N
Matsuura
S
S
D001419:Bacteria; D004731:Endotoxins / Q000032:analysis*; D005658:Fungi; D005936:Glucans; D008033:Limulus Test; D047071:beta-Glucans
D004731:Endotoxins; D005936:Glucans; D047071:beta-Glucans; C038459:curdlan; C033363:beta-1,3-glucan