COMPOSITIONS FOR DETECTING ANTIBODIES TO BABESIA MICROTI AND METHODS OF USES THEREOF

• US 20090111123A1
• Filed: 10/25/2007
• Published: 04/30/2009
• Est. Priority Date: 10/25/2007
• Status: Active Grant

• Current Assignee: Abbott Laboratories
• Original Assignee: Abbott Laboratories
• Inventors: Cheng, Yu, Birkenmeyer, Larry G., Desai, Suresh M., Shah, Dinesh O., Dawson, George J.
• Granted Patent: US 8,283,124 B2
• Time in Patent Office: Days
• US Class Current: 435/7.200
• CPC Class Codes: C07K 14/44 from protozoa
• CPC Class Codes: C07K 16/20 from protozoa
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: G01N 33/56905 Protozoa

Abstract

The present invention relates to one or more peptides which may be used to test for the presence of antibodies to B. microti. These peptide are selected based on the strength and specificity of the human antibody response to B. microti. These peptides alone or in combination have a variety of uses, for example, as a component of a diagnostic assay or kit.

Claims

1. An agent selected from the group consisting of the following:

(a) a plurality of peptides comprising a first group of peptides and an optional second group of peptides,

(i) wherein each peptide of the first group comprises a sequence as set forth in SEQ ID NOS;

5-11 or a sequence at least 70% identical thereto, and

(ii) wherein each peptide of the first group comprises no more than 50 amino acids;

(b) a chimeric protein comprising the plurality of peptides of (a); and

(c) a fusion protein comprising a first portion and a second portion, wherein the first portion is a protein according to (b) and the second portion is a binding member.

2. The agent of claim 1, wherein the first group of peptides comprises each of the sequences set forth in SEQ ID NOS:

5-11.

3. A chimeric protein according to claim 1, wherein the protein comprises the sequence of SEQ ID NO:

2.

4. A fusion protein according to claim 1, wherein the protein comprises the sequence of SEQ ID NO:

4.

5. A fusion protein according to claim 1 further comprising a third portion, wherein the third portion is a protein according to claim 1(b).

6. An isolated nucleic acid encoding at least one of said plurality of peptides of (a) of claim 1, said chimeric protein of (b) of claim 1 or said fusion protein of (c) of claim 1.

7. The isolated nucleic acid of claim 6, wherein the nucleic acid comprises the sequence of SEQ ID NO:

1.

8. The isolated nucleic acid of claim 6, wherein the nucleic acid comprises the sequence of SEQ ID NO:

3.

9. An expression vector comprising the nucleic acid of claim 6 operably linked to an expression control sequence.

10. The expression vector of claim 9, wherein said expression control sequence is a promoter.

11. An isolated host cell comprising the expression vector of claim 9.

12. A solid support comprising an agent according to claim 1.

13. The solid support of claim 12, wherein the solid support is selected from the group consisting of nitrocellulose, nylon, plastic and paper.

14. A diagnostic kit comprising an agent according to claim 1.

15. A method of detecting the presence of antibodies to B. microti in a test sample comprising the steps of:

(a) contacting said test sample with an agent according to claim 1 for a time and under conditions sufficient to form agent/antibody complexes; and

(b) detecting the presence of said agent/antibody complexes, presence of said complexes indicating presence of antibodies to B. microti in said test sample.

16. The method of claim 15, wherein said test sample is selected from the group consisting of blood, serum, plasma, saliva, cerebrospinal fluid and urine.

17. The method of claim 15, wherein the agent is attached to a solid support.

18. A method of diagnosing B. microti injection in a subject comprising the steps of:

(a) detecting the presence of antibodies to B. microti in a test sample according to the method of claim 15; and

(b) comparing the amount of anti-B. microti antibody to a control, wherein a level of anti-B microti antibody above the control is indicative of B. microti infection.

19. A method of detecting antibodies to B. microti in a test sample comprising the steps of:

(a) contacting said test sample with an agent according to claim 1 for a time and under conditions sufficient to form agent/antibody complexes;

(b) adding a conjugate to the resulting agent/antibody complexes for a time and under conditions sufficient to allow said conjugate to bind to the bound antibody, wherein said conjugate comprises an anti-antibody attached to a signal-generating compound capable of generating a detectable signal; and

(c) detecting presence of antibody which may be present in said sample by detecting a signal generated by said signal-generating compound, presence of said signal indicating presence of B. microti antibodies in said test sample.

20. The method of claim 19, wherein said test sample is selected from the group consisting of blood, serum, plasma, saliva, cerebrospinal fluid and urine.

21. The method of claim 19, wherein the agent is attached to a solid support.

22. A method of diagnosing B. microti injection in a subject comprising the steps of:

(a) detecting the presence of antibodies to B. microti in a test sample according to the method of claim 19; and

(b) comparing the amount of anti-B. microti antibody to a control, wherein a level of anti-B microti antibody above the control is indicative of B. microti infection in said subject.