大過剰のカルボキシメチル化カードランによるG因子系阻害作用を利用したエンドトキシン特異的リムルステストの開発とその応用

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  • Development of an endotoxin-specific Limulus amebocyte lysate test blocking .BETA.-glucan-mediated pathway by carboxymethylated curdlan and its application.
  • ダイカジョウ ノ カルボキシメチルカ カードラン ニ ヨル G インシケイ ソ

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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)-β-D-glucan derivative as a blocker of the (1→3)-β-D-glucan-mediated coagulation pathway. The addition of 0.1mg/ml or more of carboxymethylated (1→3)-β-D-glucan completely blocked the activation of LAL by (1→3)-β-D-glucan itself. The assay of endotoxin was unaffected by the presence of 1mg/ml carboxymethylated (1→3)-β-D-glucan. Spiked endotoxin was recovered well from β-glucans by the turbidimetric kinetic method with LAL containing 1mg/ml of carboxymethylated (1→3)-β-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.

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