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
(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 n1-n2-S-P-n5-n6-P (SEQ ID NO: 149) or a fragment thereof, wherein n1 is an amino acid selected from the group consisting of A and V, n2 is an amino acid selected from the group consisting of E and D, n5 is an amino acid selected from the group consisting of K and R, and n6 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-c2-E-G-c5-Q-Q-E-G-A-Q-Q-P-S-C(SEQ ID NO: 150) or a fragment thereof, wherein c2 is an amino acid selected from the group consisting of Q and R, and c5 is an amino acid selected from the group consisting of V and A; or
(b) A-V-c3-E-G-c6-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; and
detecting formation of an antibody-peptide complex comprising one or more isolated peptides in the mixture,
wherein formation of said complex is indicative of an antibody to an epitope of 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 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.
4. The method of claim 1 , wherein said sample is a bodily fluid.
5. The method of claim 1 , wherein said sample is from a human, canine, or equine subject.