Detection of secreted aspartyl proteases from Candida species

• US 7,745,158 B2
• Filed: 12/14/2005
• Issued: 06/29/2010
• Est. Priority Date: 12/14/2005
• Status: Expired due to Fees

• Current Assignee: Kimberly-Clark Worldwide Incorporated (Kimberly-Clark Corporation)
• Original Assignee: Kimberly-Clark Worldwide Incorporated (Kimberly-Clark Corporation)
• Inventors: DiGiammarino, Enrico L., Phillips, Erica M., McGrath, Kevin P.
• Primary Examiner(s): Mondesi; Robert
• Assistant Examiner(s): SHAHNAN SHAH, KHATOL S
• Application Number:
  US11/302,975
• Publication Number:
  US 20070134743A1
• Time in Patent Office: 1,658 Days
• Field of Search: None
• US Class Current: 435/7.31
• CPC Class Codes: C12Q 1/37 involving peptidase or proteinase
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: G01N 33/56961 Plant cells or fungi

Abstract

Methods and devices for the detection of proteins secreted by the hyphal growth form of Candida species are disclosed. The disclosed devices may constitute a method for the diagnosis of acute or chronic infections, including candidiasis, caused by microorganisms of the species Candida, such as C. albicans, for example. The devices of the present invention incorporate antibodies specific to secreted aspartyl protease proteins whose expression is upregulated upon the conversion of the Candida species from the commensal to the pathogenic form. The antibodies may be used in assays to allow the diagnosis of candidal infections and disease conditions. Either monoclonal antibodies or polyclonal antibodies may be used, and in the case of the monoclonals, the specific epitopes of the SAP protein may be detected as well as the SAP protein itself.

Claims

1. A diagnostic test kit for detecting a secreted aspartyl protease protein within a test sample, the diagnostic test kit comprising:

a lateral flow heterogeneous assay device comprising a fluidic medium, the fluidic medium defining a detection zone within which is immobilized a receptive material, wherein a test sample applied to the lateral flow assay device flows to the detection zone, and the detection zone generates a detection signal that corresponds to the presence or absence of a secreted aspartyl protease protein; and

a detection probe conjugated with a binding member;

wherein the receptive material, the binding member, or both contain a first antibody that is capable of specifically binding to the secreted aspartyl protease protein.

2. The diagnostic test kit of claim 1, wherein the receptive material contains the first antibody.

3. The diagnostic test kit of claim 2, wherein the binding member contains a second antibody that specifically binds to the secreted aspartyl protease protein.

4. The diagnostic test kit of claim 3, wherein the first antibody is a monoclonal antibody that specifically binds to a first epitope of the secreted aspartyl protease protein and the second antibody is a monoclonal antibody that specifically binds to a second epitope of the secreted aspartyl protease protein.

5. The diagnostic test kit of claim 1, wherein the binding member contains the first antibody.

6. The diagnostic test kit of claim 1, wherein the first antibody is a monoclonal antibody.

7. The diagnostic test kit of claim 1, wherein the first antibody is a polyclonal antibody.

8. The diagnostic test kit of claim 1, wherein the fluidic medium is a porous membrane.

9. The diagnostic test kit of claim 1, wherein the first antibody has been developed against a recombinant secreted aspartyl protease protein or an immunogenic fragment thereof.

10. The diagnostic test kit of claim 9, wherein the first antibody has been developed by a hybridoma cell that has been generated through fusion of a human tumor cell with a cell from an animal immunized with the recombinant secreted aspartyl protease protein or an immunogenic fragment thereof.

11. The diagnostic test kit of claim 1, wherein the diagnostic test kit is capable of detecting infection by Candida albicans.

12. The diagnostic test kit of claim 1, the lateral flow assay device further comprising an absorbent pad located downstream from the detection zone.

13. The diagnostic test kit of claim 1, the lateral flow assay device further comprising a conjugate pad located upstream of the detection zone.

14. The diagnostic test kit of claim 13, the conjugate pad carrying the detection probe in a dehydrated form, the detection probe being free to migrate upon rehydration with a test sample.

15. The diagnostic test kit of claim 1, the lateral flow assay device further defining an indicator zone.

16. The diagnostic test kit of claim 1, the lateral flow assay device further defining a calibration zone.

17. The diagnostic test kit of claim 1, wherein the conjugated detection probes or complexes thereof are separated from the test sample as the test sample flows across or through the lateral flow assay device.