Compositions for the detection of microbial contaminants

• US 10,119,969 B2
• Filed: 04/27/2011
• Issued: 11/06/2018
• Est. Priority Date: 03/17/2003
• Status: Active Grant

• Current Assignee: Charles River Laboratories Incorporated (Pathology Associates, Inc.)
• Original Assignee: Charles River Laboratories Incorporated (Pathology Associates, Inc.)
• Inventors: Wainwright, Norman R., Jordan, Foster T.
• Primary Examiner(s): Hobbs, Lisa J
• Application Number:
  US13/095,390
• Publication Number:
  US 20110201049A1
• Time in Patent Office: 2,750 Days
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2400/049 vacuum
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: G01N 2400/24 beta-D-Glucans, i.e. having beta 1,n (n=3,4,6) linkages between saccharide units, e.g. xanthan
• CPC Class Codes: G01N 2400/50 Lipopolysaccharides; LPS
• CPC Class Codes: G01N 33/579 involving limulus lysate

Abstract

The invention provides methods and compositions for the detection and/or quantification of a microbial contaminant, for example, a bacterial endotoxin or a glucan, in a sample. In particular, the invention provides a test cartridge useful in the practice of hemocyte lysate-based assays for the detection and/or quantification of a microbial contaminant in a sample. In addition, the invention provides methods of making and using such cartridges. In addition, the invention provides a rapid, sensitive, multi-step kinetic hemocyte lysate-based assay for the detection and/or quantification of a microbial contaminant in a sample. In addition, the invention provides a glucan-specific lysate that can be used in a variety of assay formats, including, for example, a test cartridge, optionally configured to perform a kinetic assay.

Claims

1. A manufacture comprising a solid support having dried thereon a composition comprising a mixture of a hemocyte lysate and (i) an anti-frothing agent, (ii) a resolubilizing agent, (iii) an anti-flaking agent and an anti-frothing agent, (iv) an anti-flaking agent and a resolubilizing agent, (v) an anti-frothing agent and a resolubilizing agent, or (vi) an anti-flaking agent, an anti-frothing agent and a resolubilizing agent, wherein said lysate is capable of being solubilized or reconstituted by a sample.

2. The manufacture of claim 1, wherein said composition further comprises a substrate.

3. The manufacture of claim 1, wherein said composition comprises a hemocyte lysate and an anti-frothing agent.

4. The manufacture of claim 1, wherein said composition comprises a hemocyte lysate and a resolubilizing agent.

5. The manufacture of claim 1, wherein said composition comprises a hemocyte lysate, an anti-flaking agent, and an anti-frothing agent.

6. The manufacture of claim 1, wherein said composition comprises a hemocyte lysate, an anti-flaking agent, and a resolubilizing agent.

7. The manufacture of claim 1, wherein said composition comprises a hemocyte lysate, an anti-frothing agent, and a resolubilizing agent.

8. The manufacture of claim 1, wherein said composition comprises a hemocyte lysate, an anti-flaking agent, an anti-frothing agent, and a resolubilizing agent.

18. The manufacture of claim 1, wherein said anti-frothing agent is polyvinyl alcohol or polypropylene glycol.

19. The manufacture of claim 1, wherein said resolubilizing agent is mannitol, mannose, sorbitol, trehalose, maltose, dextrose, sucrose, or another monosaccharide.

20. The manufacture of claim 1, wherein said resolubilizing agent is not a disaccharide.

21. The manufacture of claim 1, wherein said anti-flaking agent is selected from the group consisting of polyethylene glycol, polyvinyl pyrolidone, polyvinyl alcohol, mannitol, or a protein.

22. The manufacture of claim 1, wherein said anti-flaking agent is not dextran.

9. A manufacture comprising a solid support comprising glass or plastic having dried thereon a composition comprising mixture of a hemocyte a lysate and (i) an anti-flaking agent, (ii) an anti-frothing agent, (iii) a resolubilizing agent, (iv) an anti-flaking agent and an anti-frothing agent, (v) an anti-flaking agent and a resolubilizing agent, (vi) an anti-frothing agent and a resolubilizing agent, or (vii) an anti-flaking agent, an anti-frothing agent and a resolubilizing agent, wherein said lysate is capable of being solubilized or reconstituted by a sample.

10. The manufacture of claim 9, wherein said composition further comprises a substrate.

11. The manufacture of claim 9, wherein said composition comprises a hemocyte lysate and an anti-flaking agent.

12. The manufacture of claim 9, wherein said composition comprises a hemocyte lysate and an anti-frothing agent.

13. The manufacture of claim 9, wherein said composition comprises a hemocyte lysate and a resolubilizing agent.

14. The manufacture of claim 9, wherein said composition comprises a hemocyte lysate, an anti-flaking agent, and an anti-frothing agent.

15. The manufacture of claim 9, wherein said composition comprises a hemocyte lysate, an anti-flaking agent, and a resolubilizing agent.

16. The manufacture of claim 9, wherein said composition comprises a hemocyte lysate, an anti-frothing agent, and a resolubilizing agent.

17. The manufacture of claim 9, wherein said composition comprises a hemocyte lysate, an anti-flaking agent, an anti-frothing agent, and a resolubilizing agent.