Peptides and methods for the detection of lyme disease antibodies
The invention provides compositions (e.g., peptide compositions) useful for the detection of antibodies that bind to Borrelia antigens. The peptide compositions comprise polypeptide sequences comprising variants in the IR6 domain of the Borrelia VlsE protein. The invention also provides devices, methods, and kits comprising such peptide compositions and useful for the detection of antibodies that bind to Borrelia antigens and the diagnosis of Lyme disease.
1. A method for detecting in a sample an antibody to an epitope of a Borrelia antigen, the method comprising:
contacting a sample with a mixture of isolated peptides comprising three or more different isolated peptides, wherein each isolated peptide comprises a sequence of L-K-K-D-D-N-I-A-A-A-X11-V-L-R-G-X16-X17-K-D-G-X21-F-A-X24-X25 (SEQ ID NO: 1) wherein X11 is an amino acid selected from the group consisting of V and L, X16 is an amino acid selected from the group consisting of L and I, X17 is an amino acid selected from the group consisting of A and V, X21 is an amino acid selected from the group consisting of R, D and N, X24 is an amino acid selected from the group consisting of I, W, and Y, and X25 is an amino acid selected from the group consisting of K and R; and
detecting formation of an antibody-peptide complex comprising at least one of said isolated peptides,
wherein formation of said complex is indicative of an antibody to an epitope of a Borrelia antigen being present in said sample.
2. The method of claim 1 , wherein said Borrelia antigen is from a Borrelia burgdorferi, Borrelia afzelli , or Borrelia garinii species.
3. The method of claim 1 , wherein at least one of said isolated peptides is immobilized to a solid support.
4. The method of claim 3 , 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.
5. The method of claim 1 , 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.
6. The method of claim 1 , wherein said sample is from a human, canine, or equine subject.
7. The method of claim 1 , wherein said sample is a blood, serum, cerebral spinal fluid, urine, or saliva sample.
8. A kit comprising a mixture of isolated peptides comprising three or more different isolated peptides, wherein each isolated peptide comprises a sequence of L-K-K-D-D-N-I-A-A-A-X11-V-L-R-G-X16-X17-K-D-G-X21-F-A-X24-X25 (SEQ ID NO: 1) wherein X11 is an amino acid selected from the group consisting of V and L, X16 is an amino acid selected from the group consisting of L and I, X17 is an amino acid selected from the group consisting of A and V, X21 is an amino acid selected from the group consisting of R, D and N, X24 is an amino acid selected from the group consisting of I, W, and Y, and X25 is an amino acid selected from the group consisting of K and R and a labeling reagent capable of binding to an antibody that recognizes an epitope of at least one of said isolated peptides.
9. The kit of claim 8 , wherein at least one of said isolated peptides is attached to a solid support.
10. The kit of claim 8 , wherein at least one of said isolated peptides is attached to a bead, a tube or a well, a lateral flow assay device, or an analytical rotor.
11. The kit of claim 8 , wherein the labeling reagent is an anti-human or anti-canine IgG antibody conjugated to a detectable label.
12. The kit of claim 11 , wherein the detectable label is colloidal gold particles.