Peptides, devices, and methods for the detection of
The invention provides compositions (e.g., peptide compositions) useful for the detection of antibodies that bind to Ehrlichia antigens. The peptide compositions comprise polypeptide sequences based on an immunogenic fragment of the Ehrlichia Outer Membrane Protein 1 (OMP-1) protein. The invention also provides devices, methods, and kits comprising such peptide compositions and useful for the detection of antibodies that bind to Ehrlichia antigens and the diagnosis of monocytic ehrlichiosis.
1. A mixture of isolated peptides comprising three or more different isolated peptides, wherein each isolated peptide comprises the sequence of SEQ ID NO: 97,
F-S-A-K-E-E-X 7 -A-E-T-R-X 12 -T-F-G-L-X 17 -K-Q-Y-D-G-A-X 24 -I-X 26 -E-N-Q-V-Q-N-K-F-T-I-S-N (SEQ ID NO: 97),
wherein X 7 , X 12 , X 17 , X 24 , and X 26 is any amino acid, and wherein each isolated peptide is conjugated to a detectable label.
2. The mixture of claim 1 , wherein X 7 is K, X 12 is an amino acid selected from the group consisting of K and R, X 17 is an amino acid selected from the group consisting of E and D, X 24 is an amino acid selected from the group consisting of K and Q, and X 26 is an amino acid selected from the group consisting of E and T.
3. The mixture of claim 1 , wherein the detectable label is selected from the group consisting of an enzyme, a fluorescent label, a luminescent label, a radioactive label, and a metal-conjugated label.
4. The mixture of claim 3 , wherein the detectable label is an enzyme selected from the group consisting of horse radish peroxidase (HRP), alkaline phosphatase (ALP), and beta-galactosidase.
5. The mixture of claim 3 , wherein the detectable label is a colloidal particle, magnetic particle, or hollow gold nanosphere.
6. The mixture of claim 5 , wherein the colloidal particle is a metallic nanoparticle.
7. The mixture of claim 6 , wherein the metallic nanoparticle comprises gold, silver, platinum, copper, or composites thereof.
8. The mixture of claim 1 , wherein the detectable label is a colloidal gold particle.
9. The mixture of claim 1 , wherein one or more of the isolated peptides is conjugated to the detectable label directly.
10. The mixture of claim 1 , wherein one or more of the isolated peptides is conjugated to the detectable label indirectly.
11. The mixture of claim 10 , wherein one or more of the isolated peptides is conjugated to the detectable label through a biotin/avidin system.
12. The mixture of claim 1 , wherein one or more of the isolated peptides is conjugated to the detectable label via a cysteine residue.
13. A method for detecting in a sample an antibody to an epitope of an Ehrlichia antigen, the method comprising:
contacting a sample with the mixture of isolated peptides of claim 1 ; and
detecting formation of an antibody-peptide complex comprising one or more of said isolated peptides in the mixture,
wherein formation of said complex is indicative of an antibody to an epitope of an Ehrlichia antigen being present in said sample.
14. The method of claim 13 , wherein one or more of said isolated peptides in the mixture are immobilized to a solid support.
15. The method of claim 14 , wherein said solid support is a bead, a flow path in a lateral flow assay device, a well in a microtiter plate, or a flow path in a rotor.
16. The method of claim 13 , wherein said detecting step comprises (i) performing an ELISA assay, (ii) running a lateral flow assay, (iii) performing an agglutination assay, or (iv) running the sample through an analytical rotor.
17. The method of claim 13 , wherein said sample is from a human or canine subject.
18. The method of claim 13 , wherein said sample is a blood, serum, cerebral spinal fluid, urine, or saliva sample.
19. A kit comprising the mixture of isolated peptides of claim 1 and a device capable of detecting formation of an antibody-peptide complex comprising an antibody from a sample and one or more of said isolated peptides in the mixture.
20. The kit of claim 19 , wherein one or more of said isolated peptides in the mixture are attached to a solid support.
21. The kit of claim 19 , wherein the device is a lateral flow assay device, an analytical rotor, or an optical sensor.
22. The kit of claim 21 , wherein the device is a lateral flow immunoassay device.
23. The kit of claim 19 , wherein the device comprises a substance capable of immobilizing antibodies from a sample.
24. The kit of claim 23 , wherein the substance is anti-human or anti-dog IgG or IgM antibodies.
25. The kit of claim 23 , wherein the substance is protein A, protein G, protein L, or combinations thereof.
26. The kit of claim 23 , wherein the substance is a mixture of isolated peptides that is capable of specifically binding to an anti- Ehrlichia antibody.