Circovirus sequence associated with piglet weight loss disease (pwd)

Abstract

The genome sequences and the nucleotide sequences coding for the PWD circovirus polypeptides, such as the circovirus structural and non-strucutral polypeptides, vectors including the sequences, and cells and animals transformed by the vectors are provided. Methods for detecting the nucleic acids or polypeptides, and kits for diagnosing infection by a PWD circovirus, also are provided. Method for selecting compounds capable of modulating the viral infection are further provided. Pharmaceutical, including vaccines, compositions for preventing and/or treating viral infections caused by PWD circovirus and the use of vectors for preventing and/or treating diseases also are provided.

1 . A composition comprising a recovered recombinant protein expressed by open reading frame 2 (ORF′2) of PCVB, wherein said recombinant protein is obtained by a method comprising the steps of: a) cloning said recombinant ORF′2 from PCVB into a transfer vector; b) transferring the portion of said transfer vector containing said recombinant ORF′2 into a virus; c) infecting cells in media with said virus; d) causing said virus to express the protein from said ORF′2 and the cells to secrete said protein into the media; e) separating the cells from the media; and recovering said expressed ORF′2 protein from the media. 
     
     
         2 . A composition for inducing an immune response against PCVB comprising a protein, wherein said protein is obtained by a method comprising the steps of: a) transferring a recombinant DNA from ORF′2 of PCVB into a virus; b) infecting cells with said virus, said cells being in a growth media; c) causing said virus to express the protein from said ORF′2 and the cells to secrete said protein into the media; d) separating the cells from the media; e) recovering said recombinant protein from the media; and f) combining said recovered protein with a suitable adjuvant or other pharmaceutically acceptable carrier or excipient. 
     
     
         3 . A composition comprising a protein expressed by ORF′2 from PCVB, wherein said protein is obtained by a method comprising the steps of: a) infecting cells in growth media with a recombinant viral vector containing said ORF′2; b) causing said virus to express the protein from said ORF′2 and the cells to secrete said protein into the media; and c) recovering said expressed protein from the media. 
     
     
         4 . A method for producing a diagnostic kit for the detection of PCVB infection in a sample which comprises: a) producing the protein according to  claim 1 ,  claim 2 , or  claim 3 ; and b) packaging one or more of said proteins into a suitable container. 
     
     
         5 . A method for producing a diagnostic kit for the detection of PCVB infection in a sample which comprises: a) producing the protein according to  claim 1 ,  claim 2 , or  claim 3 ; b) packaging one or more of said proteins into a suitable container; and c) including an instruction manual within said packaging. 
     
     
         6 . An immunogenic composition effective for lessening the severity of clinical symptoms associated with PCVB infection comprising a PCVB ORF′2 protein, or an immunogenic portion thereof, wherein said immunogenic portion has at least 10 contiguous amino acids from said PCVB ORF′2 protein. 
     
     
         7 . The immunogenic composition of  claim 6 , wherein said PCVB ORF′2 protein is:
 (a) a polypeptide of SEQ ID NO:26;   (b) any polypeptide that is at least 80% homologous to the polypeptide of (a);   (c) any immunogenic portion of the polypeptides of (a) and/or (b);   (d) the immunogenic portion of (c), comprising at least 10 contiguous amino acids included in SEQ ID NO:26;   (e) a polypeptide that is encoded by a DNA comprising SEQ ID NO:25;   (f) any polypeptide that is encoded by a DNA that is at least 80% homologous to the DNA of (e);   (g) any immunogenic portion of the polypeptides encoded by the polynucleotide of (e) and/or (f); or   (h) the immunogenic portion of (g), wherein the polynucleotide coding for said immunogenic portion comprises at least 30 contiguous nucleotides of SEQ ID NO:25.   
     
     
         8 . The immunogenic composition of  claim 7 , wherein said composition further comprises an inactivated viral vector and cell culture medium. 
     
     
         9 . The immunogenic composition of  claim 8 , wherein said inactivated viral vector is a recombinant baculovirus coding for PCVB ORF′2 protein. 
     
     
         10 . The immunogenic composition of  claim 7 , wherein said composition further comprises an additional ingredient selected from the group consisting of carriers, adjuvants, media, viral inactivators, and combinations thereof. 
     
     
         11 . A container comprising at least one does of the immunogenic composition of  claim 7 . 
     
     
         12 . A kit comprising the container of  claim 11  and an instruction manual, wherein said instruction manual includes information for the intramuscular application of at least one dose of said immunogenic composition into piglets for lessening the severity of clinical symptoms associated with PCVB infection. 
     
     
         13 . The kit of  claim 12 , wherein said instruction manual further includes information for a second or further administration(s) of at least one dose of said immunogenic composition, wherein the second administration or any further administration is at least 5 weeks beyond the last administration.