Methods and compositions for the detection of bacterial endotoxins

• US 6,391,570 B1
• Filed: 09/18/2000
• Issued: 05/21/2002
• Est. Priority Date: 10/09/1997
• Status: Expired due to Term

• Current Assignee: Charles River Laboratories Incorporated (Pathology Associates, Inc.)
• Original Assignee: Charles River Laboratories Incorporated (Pathology Associates, Inc.)
• Inventors: Jordan, Foster T., Cooper, James F., Chiang, Hui-Ti, Wainwright, Norman R.
• Primary Examiner(s): Swartz, Rodney P
• Application Number:
  US09/665,221
• Time in Patent Office: 610 Days
• Field of Search: 424/184.1, 424/234.1, 424/278.1, 424/282.1, 435/7.32, 435/4, 435/7.2, 435/34, 435/184, 435/962, 514/23
• US Class Current: 435/7.32
• CPC Class Codes: G01N 33/579 involving limulus lysate
• CPC Class Codes: Y10S 435/962 Prevention or removal of interfering materials or reactants or other treatment to enhance results, e.g. determining or preventing nonspecific binding

Abstract

The invention provides methods and compositions for the detection and/or quantification of bacterial endotoxins. In particular, provided herein is an inexpensive and reproducible method for producing an improved amebocyte lysate preparation having reduced Factor G activity. Provided also is an endotoxin-specific amebocyte lysate preparation produced by such a method. In addition, the invention provides methods and compositions akin for enhancing the sensitivity to endotoxins of amebocyte lysate preparations having reducing Factor G activity. In particular, the sensitivity of such amebocyte lysate preparations to endotoxins can be enhanced by the addition of exogenous (1→3) β-D-glucan.

Claims

1. A composition comprising an amebocyte lysate preparation having reduced Factor G activity and an exogenously added (1→

3)-β

-D glucan in an amount sufficient to enhance the sensitivity of said amebocyte lysate preparation to endotoxin relative to said amebocyte lysate preparation without said exogenously added (1→

3)-β

-D glucan.

2. The composition of claim 1, wherein said (1→

3)-β

-D glucan is selected from the group consisting of cotton extract, cellulose acetate rinse, curdlan, pachyman, scleratan, leutinan, schizophyllan, coriolan, laminaran, and laminarin.

3. The composition of claim 1, wherein said (1→

3)-β

-D glucan is laminarin.

4. In a method of detecting a bacterial endotoxin in a sample, the improvement comprising using the composition of claim 1 or 3.