Proteins used for the diagnosis of lyme borreliosis

Abstract

Chimera proteins includlng: (i) at least one sequence of a DbpA protein of a Borrelia species selected from B. afzelii, B. burgdorferi sensu stricto and B. garinii, and (ii) at least one sequence of an OspC protein of a Borrelia species selected from B. afzelii, B. burgdorferi sensu stricto and B. garinii. Also, a method and a kit for the in vitro diagnosis of Lyme borreliosis using said proteins.

1 . A polypeptide comprising (i) a Decorin-binding protein A (DbpA) extracellular domain portion from  B. afzelii, B. burgdorferi sensu stricto,  or  B. garinii  that includes DbpA immunodominant epitope regions, and (ii) an Outer-surface protein C (OspC) extracellular domain portion from  B. afzelii, B. burgdorferi sensu stricto,  or  B. garinii  that includes OspC immunodominant epitope regions. 
     
     
         2 . The polypeptide as claimed in.  claim 1 , wherein the DbpA extracellular domain portion is on the N-terminal side of the polypeptide, and the OspC extracellular domain portion is on the C-terminal side of the polypeptide. 
     
     
         3 . The polypeptide as claimed in  claim 1 , further comprising a linking region between the DbpA extracellular domain portion and the OspC extracellular domain portion. 
     
     
         4 . A polypeptide comprising (i) a first amino acid sequence having at least 40% sequence identity with any of the amino acid sequences selected from the group consisting of SEQ ID NOs: 1, 3, 5, and 6, and (ii) a second amino acid sequence having at least 50% sequence identity with any of the amino acid sequences selected from the group consisting of SEQ ID NOs: 2, 4, and 7, wherein the first amino acid sequence and the second amino acid sequence are capable of forming an immunological complex with antibodies produced following a  Borrelia  infection. 
     
     
         5 . The polypeptide as claimed in  claim 4 , wherein the first amino acid sequence is on the N-terminal side of the polypeptide, and the second amino acid sequence is on the C-terminal side of the polypeptide. 
     
     
         6 . The polypeptide as claimed in.  claim 4 , further comprising a linking region between the first amino acid sequence and the second amino acid sequence. 
     
     
         7 . The polypeptide as claimed in  claim 4 , further comprising a third amino acid sequence of at least 6 contiguous histidines at the N-terminal or C-terminal end. 
     
     
         8 . A nucleic acid encoding the polypeptide as claimed in  claim 4 . 
     
     
         9 . A polypeptide comprising (i) a first amino acid sequence having at least 40% sequence identity with the amino acid sequence of SEQ ID NO: 1, and (ii) a second amino acid sequence having at least 50% sequence identity with the amino acid sequence of SEQ ID NO: 2, wherein the first amino acid sequence and the second amino acid sequence are capable of forming an immunological complex with antibodies produced following a  Borrelia  infection. 
     
     
         10 . A diagnostic kit comprising the polypeptide as claimed in  claim 1 . 
     
     
         11 . The diagnostic kit as claimed in  claim 10 , further comprising an anti-human-immunoglobulin labeled with a label. 
     
     
         12 . The diagnostic kit as claimed in  claim 10 , further comprising a chimeric Variable Lipoprotein Surface-Exposed (VlsE) polypeptide. 
     
     
         13 . A diagnostic kit comprising the polypeptide as claimed in  claim 4 . 
     
     
         14 . The diagnostic kit as claimed in  claim 13 , further comprising an anti-human-immunoglobulin labeled with a label. 
     
     
         15 . The diagnostic kit as claimed in  claim 13 , further comprising a chimeric Variable Lipoprotein Surface-Exposed (VlsE) polypeptide. 
     
     
         16 . A. method for the in vitro diagnosis of Lyme borreliosis using a biological sample, comprising:
 bringing the biological sample into contact with at least one polypeptide as claimed in  claim 1 ; and   determining whether there is formation of an immunological complex between the polypeptide and antibodies of the biological sample.   
     
     
         17 . The method as claimed in  claim 16 , wherein formation of the immunological complex is determined by adding at least one anti-human-immunoglobulin labeled with a label. 
     
     
         18 . The method as claimed in  claim 16 , wherein the polypeptide is immobilized on a solid support. 
     
     
         19 . A method for the in vitro diagnosis of Lyme borreliosis using a biological sample, comprising:
 bringing the biological sample into contact with at least one polypeptide as claimed in  claim 4 ; and   determining whether there is formation of an immunological complex between the polypeptide and antibodies of the biological sample.   
     
     
         20 . The method as claimed in  claim 19 , wherein formation of the immunological complex is determined by addina at least one anti-human-immunoglobulin labeled with a label.