Method for determining (1-3)-.beta.-D-glucan

• US 5,266,461 A
• Filed: 02/20/1992
• Issued: 11/30/1993
• Est. Priority Date: 06/21/1990
• Status: Expired due to Term

• Current Assignee: Seikagaku Kogyo Kabushiki Kaisha
• Original Assignee: Seikagaku Corporation
• Inventors: Tanaka, Shigenori
• Primary Examiner(s): Rosen, Sam
• Application Number:
  US07/835,956
• Time in Patent Office: 649 Days
• Field of Search: 435/7.21, 435/7.32, 435/7.37
• US Class Current: 435/7.21
• CPC Class Codes: G01N 33/579 involving limulus lysate

Abstract

The present invention provides a reagent for determining (1→3)-β-D-glucan comprising limulus amebocyte lysate and an antibody to an endotoxin-sensitive factor, or comprising endotoxin-sensitive factor-free lysate, which enables highly sensitive determination of mycotic (1→3)-β-D-glucan present in biological samples such as blood, urine and cerebrospinal fluid without any interference of endotoxin. Thus, deep mycosis difficult to detect can be rapidly diagnosed with a good reproducibility. Accordingly, the present invention greatly contributes to the clinical diagnosis. specifically the assay is aided by the removal of intefering endotoxin sentitive factor.

Claims

1. A method for determining (1→

3)-β

-D-glucan in a sample comprising the steps of;

(a) admixing limulus amebocyte lysate and an antibody having binding specificity for endotoxin-sensitive factor, wherein said antibody neutralizes endotoxin-sensitive factor reactivity to endotoxin; and

(b) contacting the resulting admixture with said sample, and assaying for clotting enzyme activity.

2. A method for determining (1→

3)-β

-D-glucan in a sample comprising the steps of;

(a) removing endotoxin-sensitive factor from limulus amebocyte lysate by contacting the lysate with an antibody having binding specificity for endotoxin-sensitive factor, wherein said antibody is immobilized on a carrier; and

(b) contacting the resulting endotoxin-sensitive factor-free limulus amebocyte lysate with said sample, and assaying for clotting enzyme activity.

3. A method for determining (1→

3)-β

-D-glucan in a sample comprising the steps of;

(a) adding to a sample, an antibody having binding specificity for endotoxin-sensitive factor, wherein said antibody is capable of neutralizing endotoxin-sensitive factor reactivity to endotoxin; and

(b) contacting the resulting mixture with limulus amebocyte lysate so as to neutralize endotoxin-sensitive factor reactivity to endotoxin, and assaying for clotting enzyme activity.