Peptides and methods for the detection of lyme disease antibodies

Abstract

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.

What is claimed: 
     
         1 . An isolated peptide comprising:
 (a) a VlsE 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   (b) a sequence from at least one non-VlsE  Borrelia  antigen.   
     
     
         2 . The isolated peptide of  claim 1 , wherein the VlsE sequence of 1(a) comprises the sequence of L-K-K-D-D-N-I-A-A-A-L-V-L-R-G-L-A-K-D-G-R-F-A-I-K (SEQ ID NO: 4). 
     
     
         3 . The isolated peptide of  claim 1 , wherein the sequence from at least one VlsE  Borrelia  antigen comprises a peptide sequence from the Outer Surface Protein C (OspC) of  Borrelia.    
     
     
         4 . The isolated peptide of  claim 3 , wherein the peptide sequence from the OspC is located at the N terminal of the isolated peptide. 
     
     
         5 . The isolated peptide of  claim 3 , wherein the peptide sequence from the OspC comprises a sequence of 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. 
     
     
         6 . The isolated peptide of  claim 1 , wherein the sequence from at least one VlsE  Borrelia  antigen comprises a peptide sequence from p41 of  Borrelia.    
     
     
         7 . The isolated peptide of  claim 6 , wherein the peptide sequence from p41 is located at the C terminal of the isolated peptide. 
     
     
         8 . The isolated peptide of  claim 6 , wherein the peptide sequence from p41 comprises a sequence of 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. 
     
     
         9 . The isolated peptide of  claim 6 , wherein the peptide sequence from p41 comprises a sequence of 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. 
     
     
         10 . The isolated peptide of  claim 1 , wherein the sequence from at least one non-VlsE  Borrelia  antigen includes each of: a sequence from p41 of  Borrelia  located at the C terminal of the isolated peptide and a sequence from OspC of  Borrelia  located at the N terminal of the isolated peptide wherein:
 a) the OspC sequence comprises the sequence of 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   b) the p41 sequence comprises either   i) the sequence 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   ii) the sequence 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.   
     
     
         11 . The isolated peptide of  claim 1 , wherein said isolated peptide is conjugated to a ligand. 
     
     
         12 . The isolated peptide of  claim 1 , wherein said isolated peptide is biotinylated. 
     
     
         13 . The isolated peptide of  claim 1 , wherein said isolated peptide is conjugated to streptavidin or bovine serum albumin optionally via an additional terminal amino acid linking residue. 
     
     
         14 . The isolated peptide of  claim 1 , wherein said isolated peptide is attached or immobilized to a solid support. 
     
     
         15 . The isolated peptide of  claim 1 , wherein the isolated peptide is attached or immobilized to a bead, a flow path in a lateral flow immunoassay device, a well in a microtiter plate, or a flow path in a rotor. 
     
     
         16 . A method for diagnosing Lyme disease in a subject, the method comprising:
 contacting a sample from the subject with the isolated peptide of  claim 1 ; and   detecting formation of an antibody-peptide complex comprising the isolated peptide, wherein formation of the complex is indicative the subject having Lyme disease.   
     
     
         17 . A kit comprising the isolated peptide of  claim 1  and a labeling reagent capable of binding to an antibody that recognizes an epitope of the isolated peptide. 
     
     
         18 . The kit of  claim 17 , wherein the isolated peptide is attached to a solid support. 
     
     
         19 . The kit of  claim 18 , wherein the isolated peptide is attached to a bead, a tube, a well, a lateral flow assay device, or an analytical rotor. 
     
     
         20 . The kit of  claim 17 , wherein the labeling reagent is (i) an anti-human or anti-canine IgG antibody conjugated to a detectable label or (ii) protein A and/or protein G conjugated to a detectable label. 
     
     
         21 . The kit of  claim 20  wherein the detectable label is colloidal gold nanoparticles.