Hepatitis a virus nucleotide sequences, recombinant proteins and uses thereof

Abstract

Hepatitis A virus primers and probes derived from the capsid proteins and junction between the capsid precursor P1 and 2A of the HAV genome are disclosed. Also disclosed are nucleic acid-based assays using the primers and probes, antigen detection of HAV, and immunoassay for detecting the antibodies that bind to HAV.

1 . A method for detecting antibodies in a biological sample that bind to a hepatitis A virus (HAV) antigen, said method comprising: 
 (a) providing an antigen comprising a sequence having at least 80% sequence identity to any one of the sequences of SEQ ID NOS:40-48, or an immunogenic fragment of any one of the sequences of SEQ ID NOS:40-48 comprising at least 10 amino acids;    (b) incubating said antigen with said biological sample under conditions that allow for formation of an antibody-antigen complex if HAV antibodies are present in said biological sample; and    (c) detecting the presence or absence of said antibody-antigen complexes.    
     
     
         2 . The method of  claim 1 , wherein said antigen comprises the sequence of SEQ ID NO:40, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         3 . The method of  claim 1 , wherein said antigen comprises the sequence of SEQ ID NO:41, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         4 . The method of  claim 1 , wherein said antigen comprises the sequence of SEQ ID NO:42, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         5 . The method of  claim 1 , wherein said antigen comprises the sequence of SEQ ID NO:43, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         6 . The method of  claim 1 , wherein said antigen comprises the sequence of SEQ ID NO:44, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         7 . The method of  claim 1 , wherein said antigen comprises the sequence of SEQ ID NO:45, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         8 . The method of  claim 1 , wherein said antigen comprises the sequence of SEQ ID NO:46, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         9 . The method of  claim 1 , wherein said antigen comprises the sequence of SEQ ID NO:47, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         10 . The method of  claim 1 , wherein said antigen comprises the sequence of SEQ ID NO:48, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         11 . The method of  claim 1 , wherein said biological sample is human blood, plasma or serum.  
     
     
         12 . The method of  claim 1 , wherein said antigen in step (a) is immobilized on a solid support.  
     
     
         13 . The method of  claim 1 , wherein said antigen comprises a sequence having at least 90% sequence identity to any one of the sequences of SEQ ID NOS :40-48, or an immunogenic fragment thereof.  
     
     
         14 . The method of  claim 1 , wherein said antigen comprises a sequence having at least 95% sequence identity to any one of the sequences of SEQ ID NOS:40-48, or an immunogenic fragment thereof.  
     
     
         15 . The method of  claim 1 , wherein said antigen comprises a sequence having at least 99% sequence identity to any one of the sequences of SEQ ID NOS:40-48, or an immunogenic fragment thereof.  
     
     
         16 . A method for detecting antibodies in a human blood, plasma or serum sample that bind to a hepatitis A virus (HAV) antigen, said method comprising: 
 (a) providing an antigen immobilized on a solid support, said antigen comprising any one of the sequences of SEQ ID NOS:40-48, or an immunogenic fragment of any one of the sequences of SEQ ID NOS:40-48 comprising at least 10 amino acids;    (b) incubating said antigen with said sample under conditions that allow for formation of an antibody-antigen complex if HAV antibodies are present in said sample; and    (c) detecting the presence or absence of said antibody-antigen complexes.    
     
     
         17 . The method of  claim 16 , wherein said antigen comprises the sequence of SEQ ID NO:40, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         18 . The method of  claim 16 , wherein said antigen comprises the sequence of SEQ ID NO:41, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         19 . The method of  claim 16 , wherein said antigen comprises the sequence of SEQ ID NO:42, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         20 . The method of  claim 16 , wherein said antigen comprises the sequence of SEQ ID NO:43, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         21 . The method of  claim 16 , wherein said antigen comprises the sequence of SEQ ID NO:44, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         22 . The method of  claim 16 , wherein said antigen comprises the sequence of SEQ ID NO:45, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         23 . The method of  claim 16 , wherein said antigen comprises the sequence of SEQ ID NO:46, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         24 . The method of  claim 16 , wherein said antigen comprises the sequence of SEQ ID NO:47, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         25 . The method of  claim 16 , wherein said antigen comprises the sequence of SEQ ID NO:48, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         26 . A solid support comprising an immobilized antigen, wherein said antigen comprises a sequence having at least 80% sequence identity to any one of the sequences of SEQ ID NOS:40-48, or an immunogenic fragment of any one of the sequences of SEQ ID NOS:40-48 comprising at least 10 amino acids.  
     
     
         27 . The solid support of  claim 26 , wherein said antigen comprises the sequence of SEQ ID NO:40, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         28 . The solid support of  claim 26 , wherein said antigen comprises the sequence of SEQ ID NO:41, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         29 . The solid support of  claim 26 , wherein said antigen comprises the sequence of SEQ ID NO:42, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         30 . The solid support of  claim 26 , wherein said antigen comprises the sequence of SEQ ID NO:43, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         31 . The solid support of  claim 26 , wherein said antigen comprises the sequence of SEQ ID NO:44, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         32 . The solid support of  claim 26 , wherein said antigen comprises the sequence of SEQ ID NO:45, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         33 . The solid support of  claim 26 , wherein said antigen comprises the sequence of SEQ ID NO:46, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         34 . The solid support of  claim 26 , wherein said antigen comprises the sequence of SEQ ID NO:47, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         35 . The solid support of  claim 26 , wherein said antigen comprises the sequence of SEQ ID NO:48, or an immunogenic fragment thereof comprising at least 10 amino acids.  
     
     
         36 . An immunodiagnostic test kit for detecting hepatitis A virus (HAV) infection, said test kit comprising: 
 (a) an antigen comprising a sequence having at least 80% sequence identity to any one of the sequences of SEQ ID NOS:40-48, or an immunogenic fragment of any one of the sequences of SEQ ID NOS:40-48 comprising at least 10 amino acids; and    (b) instructions for conducting the immunodiagnostic test.    
     
     
         37 . An immunodiagnostic test kit for detecting hepatitis A virus (HAV) infection, said test kit comprising: 
 (a) the solid support of  claim 26;  and    (b) instructions for conducting the immunodiagnostic test.    
     
     
         38 . A method for detecting hepatitis A virus (HAV) in a biological sample, said method comprising: 
 (a) providing an antibody against an antigen that comprises a sequence having at least 80% sequence identity to any one of the sequences of SEQ ID NOS:40-48, or an immunogenic fragment of any one of the sequences of SEQ ID NOS:40-48 comprising at least 10 amino acids;    (b) incubating said antibody with said biological sample under conditions that allow for formation of an antibody-antigen complex if HAV is present in said biological sample; and    (c) detecting the presence or absence of said antibody-antigen complexes.