Circovirus sequences associated with piglet weight loss disease (PWD)
View Patent ↗The genome sequences and the nucleotide sequences coding for the PWD circovirus polypeptides, such as the circovirus structural and non-structural 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 vaccine, 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 method of detecting the presence of at least one antibody directed against porcine circovirus type B (PCVB) in a sample, the method comprising:
contacting the sample with a PCVB polypeptide comprising a PCVB antigen or epitope under conditions that allow the formation of an antigen-antibody complex between said polypeptide and a PCVB antibody present in the sample, wherein the polypeptide is a polypeptide having at least 90% identity to the amino acid sequence of SEQ ID NO: 26, or encoded by a nucleotide sequence with at least 90% identity to SEQ ID NO 25; and
detecting the antigen-antibody complex formed.
2. The method of claim 1 , wherein the sample is from a pig.
3. The method of claim 1 , wherein the sample is serum, whole blood, or biopsies.
4. The method of claim 2 , wherein the method comprises:
providing a well of a microtiter plate, wherein the well comprises the PCVB polypeptide;
introducing into the well the pig sample under conditions that allow the formation of the antigen-antibody complex;
subsequently introducing into the well an enzyme-labeled antibody directed against pig immunoglobulin wherein the enzyme is capable of hydrolyzing a substrate;
adding the substrate to the well; and
detecting hydrolyzed substrate, thereby detecting the presence of PCVB antibody in the pig sample.
5. The method of claim 1 , wherein the detecting of the antigen-antibody complex formed is by the ELISA technique, by immunofluorescence, or by radioimmunological processes (RIA) or their equivalent.
6. The method of claim 1 , wherein the polypeptide is selected from the group consisting of SEQ ID NO: 29, SEQ. ID NO: 30, SEQ ID NO: 31 and SEQ ID NO: 32.
7. The method of claim 1 , wherein the polypeptide is labeled with a detectable marker.
8. A kit for detecting at least one antibody directed against porcine circovirus type B (PCVB) in a sample, the kit comprising:
a PCVB polypeptide comprising a PCVB antigen or epitope, wherein the polypeptide has at least 90% identity to the amino acid sequence of SEQ ID NO:26, or is encoded by a nucleotide sequence with at least 90% identity to SEQ ID NO:25; and
at least one reagent that allows the detection of an antigen-antibody complex between the PCVB polypeptide and the PCVB antibody present in a sample, wherein said at least one reagent is labeled, or is able to be recognized by a labeled second reagent.
9. The kit of claim 8 , wherein the PCVB polypeptide is labeled with a detectable marker.
10. The kit of claim 8 , wherein the at least one reagent comprises a labeled antibody directed against pig immunoglobulin.
11. The kit of claim 8 , further comprising a negative control reference sample.
12. The kit of claim 8 , further comprising a reference sample containing a predetermined quantity of an antibody recognized by the PCVB polypeptide.
13. The method of claim 1 , wherein the PCVB polypeptide comprises a PCVB specific antigen or epitope for PCV type B discrimination.
14. The method f claim 1 , wherein the polypeptide SEQ ID NO: 123.