Peptides and methods for the detection of Lyme disease antibodies

1. A mixture of isolated peptides comprising three or more different isolated peptides, wherein each isolated peptide comprises

(i) a sequence according to L-K-K-D-D-N-I-A-A-A-X 11 -V-L-R-G-X 16 -X 17 -K-D-G-X 21 -F-A-X 24 -X 25 (SEQ ID NO: 1) wherein X 11 is an amino acid selected from the group consisting of V and L, X 16 is an amino acid selected from the group consisting of L and I, X 17 is an amino acid selected from the group consisting of A and V, X 21 is an amino acid selected from the group consisting of R, D and N, X 24 is an amino acid selected from the group consisting of I, W, and Y, and X 25 is an amino acid selected from the group consisting of K and R;

(ii) an N-terminal peptide sequence wherein the sequence is n 1 -n 2 -S-P-n 5 -n 6 -P (SEQ ID NO: 149) or a fragment thereof, wherein n 1 is an amino acid selected from the group consisting of A and V, n 2 is an amino acid selected from the group consisting of E and D, n 5 is an amino acid selected from the group consisting of K and R, and n 6 is an amino acid selected from the group consisting of K and R; and

(iii) a C-terminal peptide sequence wherein the sequence is

(a) V-c 2 -E-G-c 5 -Q-Q-E-G-A-Q-Q-P-S-C (SEQ ID NO: 150) or a fragment thereof, wherein c 2 is an amino acid selected from the group consisting of Q and R, and c 5 is an amino acid selected from the group consisting of V and A; or

(b) A-V-c 3 -E-G-c 6 -Q-Q-E-G-A-Q-Q-P-S (SEQ ID NO: 151) or a fragment thereof, wherein C3 is an amino acid selected from the group consisting of Q and R, and C6 is an amino acid selected from the group consisting of V and A.

2. The mixture of claim 1 , wherein each isolated peptide is conjugated to a ligand or wherein one or more of the isolated peptides is biotinylated.

3. The mixture of claim 1 , wherein one or more of the isolated peptides is conjugated to:

(i) streptavidin, polymeric streptavidin, avidin, or neutravidin;

(ii) a carrier protein selected from serum albumin, keyhole limpet hemocyanin (KLH), and an immunoglobulin Fc domain;

(iii) a dendrimer;

(iv) part of a multiple antigenic peptides system (MAPS); or

(v) a colloidal particle, nanoparticle, colloidal nanoparticle, core-shell structure particle, or hollow nanosphere.

4. The mixture of claim 1 , wherein each isolated peptide is immobilized to a solid support.

5. The mixture of claim 4 , wherein each isolated peptide is immobilized to the solid support through a terminal amino acid linking residue.

6. The mixture of claim 4 , wherein the solid support is a bead, a flow path in a lateral flow immunoassay device, a well in a microtiter plate, or a flow path in a rotor.

7. A method for diagnosing Lyme disease in a subject, the method comprising: contacting a sample from the subject with the mixture of isolated peptides claim 1 ; and detecting formation of an antibody-peptide complex comprising one or more isolated peptides in the mixture, wherein formation of the complex is indicative of the subject having Lyme disease.

8. The method of claim 7 wherein said detecting step comprises (i) performing an ELISA assay, (ii) running a lateral flow assay, (iii) performing an agglutination assay, (iv) performing a Western blot, a slot blot, or dot blot, or (v) running the sample through an analytical rotor.

9. The method of claim 7 , wherein said sample is a bodily fluid.

10. The method of claim 7 , wherein said sample is from a human, canine, or equine subject.

11. A kit comprising the mixture of isolated peptides of claim 1 and a labeling reagent capable of binding to an antibody that recognizes an epitope of one or more isolated peptides in the mixture.

12. The kit of claim 11 , wherein the labeling reagent is:

(i) an anti-human, anti-canine, or anti-equine IgG/IgM antibody conjugated to a detectable label; or

(ii) protein A and/or protein G conjugated to a detectable label.

13. The kit of claim 12 , wherein the detectable label is colloidal gold particles.