Nucleic acids of rochalimaea henselae and methods and compositions for diagnosing rochalimaea henselae and rochalimaea quintana infection

Abstract

A method of diagnosing cat scratch disease and a method of diagnosing bacillary angiomatosis in a subject by detecting the presence of Rochalimaea henselae or an antigenic fragment thereof in the subject is provided. Also provided is a vaccine comprising an immunogenic amount of a nonpathogenic Rochalimaea henselae or an immunogenically specific determinant thereof and a pharmaceutically acceptable carrier. A method of diagnosing Rochalimaea quintana infection in a subject by detecting the presence of a nucleic acid specific to Rochalimaea quintana in a sample from the subject is provided. A purified, 60-kDa heat shock protein of Rochalimaea is provided. Also provided is a 17-kDa antigenic polypeptide of Rochalimaea.

What is claimed is:  
     
         1 . An isolated nucleic acid encoding the amino acid sequence set forth in the Sequence Listing as SEQ ID NO:11.  
     
     
         2 . The isolated nucleic acid of  claim 1  in a vector suitable for expressing the nucleic acid.  
     
     
         3 . The vector of  claim 2  in a host suitable for expressing the nucleic acid.  
     
     
         4 . An isolated nucleic acid, of at least 15 nucleotides in length, which can selectively hybridize with the nucleic acid of  claim 1 , under polymerase chain reaction conditions.  
     
     
         5 . An isolated nucleic acid, of at least 15 nucleotides in length, which can specifically hybridize with the nucleic acid of  claim 1 , under the stringency conditions of 60° C. and 5×SSC, followed by the initial washing condition of room temperature, 2×SSC and 0.1% SDS, and two secondary washes with stringency conditions of 50° C., 0.5% SSC and 0.1% SDS.  
     
     
         6 . A purified antigenic polypeptide encoded by the isolated nucleic acid of  claim 5 .  
     
     
         7 . A purified antigenic polypeptide having the amino acid sequence set forth in the Sequence Listing as SEQ ID NO:11.  
     
     
         8 . A method of diagnosing current or previous cat scratch disease in a subject, comprising detecting the presence of  Rochalimaea henselae  in a sample from the subject, and correlating the presence of  Rochalimaea henselae  with cat scratch disease in the subject.  
     
     
         9 . The method of  claim 8 , wherein the detecting step comprises the steps of contacting a fluid or tissue sample from the subject with a detectable amount of a purified antibody that specifically binds  Rochalimaea henselae,  and detecting the binding of the antibody to  Rochalimaea henselae.    
     
     
         10 . The method of  claim 9 , wherein the antibody is an antibody which specifically binds to the polypeptide set forth in the Sequence Listing as SEQ ID NO:11.  
     
     
         11 . The method of  claim 9 , wherein the antibody is an antibody which specifically binds to the polypeptide set forth in the Sequence Listing as SEQ ID NO:7.  
     
     
         12 . A method of diagnosing current or previous cat scratch disease in a subject by detecting the presence of an antibody that specifically binds  R. henselae,  wherein the detecting step comprises the steps of contacting an antibody-containing fluid or tissue sample from the subject with an amount of purified  Rochalimaea henselae  or an antigenic fragment thereof which binds to the antibody, detecting the binding of the  Rochalimaea henselae  or antigenic fragment thereof to the antibody, and correlating the presence of an antibody that specifically binds to  Rochalimaea henselae  or an antigenic fragment thereof with cat scratch disease in the subject.  
     
     
         13 . The method of  claim 12 , wherein the antigenic fragment is the antigenic polypeptide encoded by the nucleic acid which can specifically hybridize with the nucleic acid encoding the amino acid sequence set forth in the Sequence Listing as SEQ ID NO:11 under the stringency conditions of 60° C. and 5×SSC, followed by the initial washing condition of room temperature, 2×SSC and 0.1% SDS, and two secondary washes with stringency conditions of 50° C., 0.5% SSC and 0.1% SDS.  
     
     
         14 . The method of  claim 12 , wherein the antigenic fragment is the polypeptide set forth in the Sequence Listing as SEQ ID NO:11.  
     
     
         15 . The method of  claim 8 , wherein the presence of  Rochalimaea henselae  is determined by detecting the presence of a nucleic acid specific for  Rochalimaea henselae.    
     
     
         16 . The method of  claim 15 , wherein the nucleic acid detected is a nucleic acid encoding the amino acid sequence set forth in the Sequence Listing as SEQ ID NO:11.  
     
     
         17 . The method of  claim 15 , wherein the nucleic acid is detected utilizing a nucleic acid amplification technique.  
     
     
         18 . The method of  claim 8 , wherein the subject is a human.  
     
     
         19 . A method of diagnosing bacillary angiomatosis in a subject comprising detecting the presence of  Rochalimaea henselae  in a sample from the subject, and correlating the presence of  Rochalimaea henselae  with bacillary angiomatosis in the subject.  
     
     
         20 . The method of  claim 19 , wherein the detecting step comprises the steps of contacting a fluid or tissue sample from the subject with a detectable amount of a purified antibody that specifically binds  Rochalimaea henselae,  and detecting the binding of the antibody to  Rochalimaea henselae.    
     
     
         21 . A method of diagnosing bacillary angiomatosis in a subject by detecting the presence of an antibody that specifically binds  Rochalimaea henselae,  wherein the detecting step comprises the steps of contacting an antibody-containing fluid or tissue sample from the subject with an amount of purified  Rochalimaea henselae  or antigenic fragment thereof which binds to the antibody, and detecting the binding of the  Rochalimaea henselae  or antigenic fragment thereof to the antibody.  
     
     
         22 . The method of  claim 19 , wherein the presence of  Rochalimaea henselae  is determined by detecting the presence of a nucleic acid specific for  Rochalimaea henselae.    
     
     
         23 . The method of  claim 19 , wherein the subject is a human.  
     
     
         24 . A nonpathogenic  Rochalimaea henselae  or an immunogenic fragment thereof.  
     
     
         25 . A purified  Rochalimaea henselae  bound to a ligand.  
     
     
         26 . The purified  Rochalimaea henselae  of  claim 25 , wherein the ligand is an antibody.  
     
     
         27 . A purified  Rochalimaea henselae  bound to a substrate.  
     
     
         28 . A purified antibody or antigen binding fragment thereof that specifically binds  Rochalimaea henselae.    
     
     
         29 . The antibody of  claim 28 , wherein the antibody is a monoclonal antibody.  
     
     
         30 . The antibody of  claim 28 , bound to a substrate.  
     
     
         31 . The antibody of  claim 28 , wherein the antibody is labeled with a detectable moiety.  
     
     
         32 . The antibody of  claim 31 , wherein the detectable moiety is fluorescent.  
     
     
         33 . A diagnostic kit for detecting the presence of a primary antibody specifically reactive with  Rochalimaea henselae  or an immunogenic fragment thereof comprising: a purified  Rochalimaea henselae  or immunogenic fragment thereof bound to a substrate; a secondary antibody reactive with the antibody specifically reactive with  Rochalimaea henselae  or an immunogenic fragment thereof; and a reagent for detecting a reaction of the secondary antibody with the primary antibody.  
     
     
         34 . An isolated immunogenically specific determinant of  Rochalimaea henselae.    
     
     
         35 . The method of  claim 12 , wherein the antigenic fragment is the polypeptide set forth in the Sequence Listing as SEQ ID NO:7.  
     
     
         36 . The method of  claim 15 , wherein the nucleic acid specific for  Rochalimaea henselae  comprises the nucleotides in the nucleotide sequence defined in the Sequence Listing as SEQ ID NO:5.  
     
     
         37 . The method of  claim 22 , wherein the nucleic acid specific for  Rochalimaea henselae  comprises the nucleotides in the nucleotide sequence defined in the Sequence Listing as SEQ ID NO:5.  
     
     
         38 . An isolated nucleic acid specific for  Rochalimaea henselae,  comprising the nucleotides in the nucleotide sequence defined in the Sequence Listing as SEQ ID NO:5.  
     
     
         39 . An isolated nucleic acid consisting of the nucleotides in the nucleotide sequence defined in the Sequence Listing as SEQ ID NO:7.  
     
     
         40 . The nucleic acid of  claim 39  in a vector suitable for expressing the nucleic acid.  
     
     
         41 . The vector of  claim 40  in a host suitable for expressing the nucleic acid.  
     
     
         42 . A purified antigenic protein encoded by the nucleic acid of  claim 39 .  
     
     
         43 . An isolated nucleic acid that specifically hybridizes with the nucleic acid of  claim 39  under high stringency conditions and at least 85% sequence complementarity with the segment of the strand to which it hybridizes.  
     
     
         44 . An antigenic polypeptide fragment encoded by the nucleic acid of  claim 39 .  
     
     
         45 . A method of diagnosing bacillary angiomatosis in a subject comprising detecting the presence of  Rochalimaea quintana  in a sample from the subject by detecting the presence of a nucleic acid specific for  Rochalimaea quintana,  wherein the nucleic acid specific for  Rochalimaea quintana  consists of the nucleotides in the nucleotide sequence defined in the Sequence Listing as SEQ ID NO:6, and correlating the presence of  Rochalimaea quintana  with the presence of bacillary angiomatosis in the subject.  
     
     
         46 . A method of diagnosing bacillary angiomatosis in a subject comprising detecting the presence of  Rochalimaea quintana  in a sample from the subject by detecting the presence of a nucleic acid specific for  Rochalimaea quintana,  wherein the nucleic acid specific for  Rochalimaea quintana  consists of the nucleotides in the nucleotide sequence defined in the Sequence Listing as SEQ ID NO:1, and correlating the presence of  Rochalimaea quintana  with the presence of bacillary angiomatosis in the subject.  
     
     
         47 . An isolated nucleic acid specific for  Rochalimaea quintana,  comprising the nucleotides in the nucleotide sequence defined in the Sequence Listing as SEQ ID NO:6.  
     
     
         48 . A method of diagnosing cat scratch disease or bacillary angiomatosis in a subject, comprising: amplifying DNA from the subject using a primer mixture consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4; contacting the amplified DNA from step a. with a probe consisting of the nucleic acid of SEQ ID NO:5 and detecting the hybridization of the probe with the amplified DNA, the existence of hybridization indicating the presence of  R. henselae,  which is correlated with cat scratch disease; and contacting the amplified DNA from step a. with a probe consisting of the nucleic acid of SEQ ID NO:6 and detecting the hybridization of the probe with the amplified DNA, the existence of hybridization indicating the presence of  R. quintana,  which is correlated with bacillary angiomatosis.  
     
     
         49 . A mixture of the nucleic acids of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4.