Compositions and Methods for Detection of Antibodies Specific for Anaplasma phagocytophilum (Aph) and Anaplasma platys (Apl)

Abstract

The invention provides methods and compositions for the detection and treatment of Anaplasma phagocytophilum and Anaplasma platys infection.

We claim: 
     
         1 . A composition comprising one or more purified polypeptides, wherein the one or more polypeptides:
 (a) consist of an amino acid sequence set forth as SEQ ID NO:13 or 21, or   (b) consist of at least 30 contiguous amino acids of an amino acid sequence set forth as SEQ ID NO:13; consist of at least 175 contiguous of an amino acid sequence set forth as SEQ ID NO:21, wherein the one or more purified polypeptides retain an N-terminal C amino acid, or   (c) consist of a polypeptide fragment having 94, 95, 96, 97, 98 or 99% identity to SEQ ID NO:2 or SEQ ID NO:10 over the full length of SEQ ID NO:2 or SEQ ID NO:10, wherein the sequence of the one or more purified polypeptides is non-naturally occurring; or   (d) consist of a polypeptide fragment having at least about 94% identity to SEQ ID NO:13 or 21 over the full length of SEQ ID NO:13 or 21, wherein the sequence of the one or more purified polypeptides is non-naturally occurring; or   (e) comprise an amino acid sequence set forth as SEQ ID NO:13, wherein the one or more polypeptides have less than 50 contiguous naturally occurring  Anaplasma  amino acids, wherein the sequence of the one or more purified polypeptides is non-naturally occurring;   wherein, optionally, the one or more purified polypeptides are linked to an indicator reagent, a signal sequence, a stop transfer sequence, an amino acid spacer, a transmembrane domain, a protein purification ligand, a heterologous polypeptide, a moiety that enhances an immune response, a moiety that facilitates purification, a moiety that facilitates polypeptide stability, one or more additional polypeptides comprising any one of SEQ ID NOs:1-21 or a combination thereof.   
     
     
         2 . The composition of  claim 1 , further comprising a pharmaceutically-acceptable or veterinarily acceptable carrier. 
     
     
         3 . The composition of  claim 1 , wherein the one or more purified polypeptides are attached to a solid support. 
     
     
         4 . The composition of  claim 1 , wherein the one or more purified polypeptides are chemically synthesized. 
     
     
         5 . The composition of  claim 1 , further comprising an adjuvant. 
     
     
         6 . A method of inducing an immune response in a mammal comprising administering an immunologically effective amount of the composition of  claim 5  to the mammal. 
     
     
         7 . A method of detecting antibodies that specifically bind an  Anaplasma platys  polypeptide or an  Anaplasma phagocytophilum  polypeptide in a test sample, comprising:
 (a) contacting the composition of  claim 1  with the test sample, under conditions that allow polypeptide/antibody complexes to form;   (b) detecting the polypeptide/antibody complexes;   wherein the detection of the polypeptide/antibody complexes is an indication that antibodies specific for an  Anaplasma platys  polypeptide or an  Anaplasma phagocytophilum  polypeptide are present in the test sample.   
     
     
         8 . The method of  claim 7 , further comprising contacting the complexes of step (a) with an indicator reagent prior to the performance of step (b). 
     
     
         9 . The method of  claim 7 , wherein the amount of antibodies in the test sample is determined. 
     
     
         10 . The method of  claim 7 , wherein the one or more purified polypeptides are attached to a substrate. 
     
     
         11 . The composition of  claim 3 , wherein the one or more purified polypeptides are also bound to an antibody that is specific for  Anaplasma platys  or  Anaplasma phagocytophilum.    
     
     
         12 . A device comprising one or more purified polypeptides, wherein the one or more polypeptides consist of an amino acid sequence set forth as SEQ ID NO:2, 10, 13, 21, or consist of at least 30 contiguous amino acids of an amino acid sequence set forth as SEQ ID NOs:2 or 13, or consist of at least 175 contiguous amino acids of an amino acid sequence set forth as SEQ ID NOs:10 or 21, or consist of a polypeptide fragment having at least about 94% identity to any one of SEQ ID NOs:2, 10, 13, or 21 over the full length of SEQ ID NOs:2, 10, 13, or 21; or wherein the one or more polypeptides comprise an amino acid sequence set forth as SEQ ID NO:2, 10, 13, or 21; wherein the one or more purified polypeptides are bound to:
 (a) a solid support; and   (b) an antibody that is specific for  Anaplasma platys  or  Anaplasma phagocytophilum;      
       wherein, optionally, the one or more purified polypeptides are linked to an indicator reagent, a signal sequence, a stop transfer sequence, an amino acid spacer, a transmembrane domain, a protein purification ligand, a heterologous polypeptide, a moiety that enhances an immune response, a moiety that facilitates purification, a moiety that facilitates polypeptide stability, one or more additional polypeptides comprising any one of SEQ ID NOs:1-21 or a combination thereof. 
     
     
         13 . A composition comprising one or more purified polypeptides, wherein the one or more polypeptides consist of an amino acid sequence set forth as SEQ ID NO:2, 10, 13, or 21, or consist of at least 30 contiguous amino acids of an amino acid sequence set forth as SEQ ID NOs:2 or 13, or consist of at least 175 contiguous amino acids of an amino acid sequence set forth as SEQ ID NOs:10 or 21, or consist of a polypeptide fragment having at least about 94% identity to any one of SEQ ID NOs:2, 10, 13, or 21 over the full length of SEQ ID NOs:2, 10, 13, or 21; or wherein the one or more polypeptides comprise an amino acid sequence set forth as SEQ ID NO:2, 10, 13, or 21;
 wherein, the one or more purified polypeptides are linked to an indicator reagent, a signal sequence, a stop transfer sequence, an amino acid spacer, a transmembrane domain, a protein purification ligand, a heterologous polypeptide, a moiety that enhances an immune response, a moiety that facilitates purification, a moiety that facilitates polypeptide stability, one or more additional polypeptides comprising any one of SEQ ID NOs:1-21 or a combination thereof.   
     
     
         14 . The composition of  claim 13 , further comprising a pharmaceutically-acceptable or veterinarily acceptable carrier. 
     
     
         15 . The composition of  claim 13 , wherein the one or more purified polypeptides are attached to a solid support. 
     
     
         16 . The composition of  claim 13 , wherein the one or more purified polypeptides are chemically synthesized. 
     
     
         17 . The composition of  claim 13 , further comprising an adjuvant. 
     
     
         18 . The composition of  claim 13 , wherein the one or more purified polypeptides are also bound to an antibody that is specific for  Anaplasma platys  or  Anaplasma phagocytophilum.    
     
     
         19 . A method of inducing an immune response in a mammal comprising administering an immunologically effective amount of the composition of  claim 13  to the mammal. 
     
     
         20 . A method of detecting antibodies that specifically bind an  Anaplasma platys  polypeptide or an  Anaplasma phagocytophilum  polypeptide in a test sample, comprising:
 (a) contacting the composition of  claim 13  with the test sample, under conditions that allow polypeptide/antibody complexes to form;   (b) detecting the polypeptide/antibody complexes;   wherein the detection of the polypeptide/antibody complexes is an indication that antibodies specific for an  Anaplasma platys  polypeptide or an  Anaplasma phagocytophilum  polypeptide are present in the test sample.   
     
     
         21 . The method of  claim 20 , further comprising contacting the complexes of step (a) with an indicator reagent prior to the performance of step (b). 
     
     
         22 . The method of  claim 20 , wherein the amount of antibodies in the test sample is determined. 
     
     
         23 . The method of  claim 20 , wherein the one or more purified polypeptides are attached to a substrate.