Methods and compositions for detecting herpes simplex virus type 2
The invention provides methods for sensitive and specific detection of anti-HSV-2 antibodies by depletion of cross-reactive (non-specific) antibodies in a biological sample that can lead to a false positive result. The invention also features compositions, including nucleic acids, polypeptides, and kits, for use in the methods of the invention.
1. A method of detecting the presence or absence of a specific anti-herpes simplex virus type-2 (HSV-2) antibody in a biological sample, comprising:
(a) contacting a biological sample suspected of containing anti-HSV-2 antibodies with a first antigen comprising the amino acid sequence GHTNTSSAS (SEQ ID NO:07), wherein said first antigen is not a full length glycoprotein G2 (gG2) polypeptide,
(b) contacting said biological sample with a second antigen capable of binding said specific anti-HSV-2 antibody, and
(c) detecting the presence or absence of specific anti-HSV-2 antibody in said biological sample wherein said specific anti-HSV-2 antibody detected in said sample is bound to said second antigen,
wherein said first antigen is not reactive with said specific anti-HSV-2 antibody.
2. The method of claim 1 , wherein said first antigen is J24, JA, JA1, or JA2.
3. The method of claim 1 , wherein step (a) further comprises contacting said biological sample with a third antigen comprising the amino acid sequence AAKTPPTTPAP (SEQ ID NO:06), wherein said third antigen is not a full length glycoprotein G2 (gG2) polypeptide, wherein said third antigen is not reactive with said specific anti-HSV-2 antibody.
4. The method of claim 1 , wherein said first antigen comprises a polypeptide having an amino acid sequence consisting essentially of GHTNTSSAS (SEQ ID NO:07).
5. The method of claim 1 , wherein said detecting further comprises contacting said sample with one or more detectably labeled anti-human immunoglobulin antibodies.
6. The method of claim 1 , wherein at least one of said antigens is immobilized on a nitrocellulose membrane support.
7. The method of claim 1 , wherein at least one of said antigens is immobilized on a solid support.
8. The method of claim 7 , wherein said solid support is selected from the group consisting of a microparticle, an agarose bead, and a magnetic bead.
9. The method of claim 1 , wherein said first antigen is not immobilized on a solid support and said second antigen is immobilized on a solid support.
10. The method of claim 1 , wherein said first antigen is immobilized on a first solid support, and said second antigen is immobilized on a second solid support.
11. The method of claim 1 , wherein said antibodies bound to said first antigen are removed from the sample prior to step (c).
12. The method of claim 1 , wherein said antibodies bound to said first antigen are removed from the sample prior to step (b).
13. The method of claim 1 , wherein said first antigen is detectably labeled.
14. The method of claim 1 , wherein said second antigen is a glycoprotein selected from the group consisting of: gC2, gG2, gB2, gD2 and fragments thereof.
15. The method of claim 1 , wherein said second antigen comprises an epitope from the glycoprotein, gG2.
16. The method of claim 1 , wherein said second antigen is the full-length glycoprotein gG2.
17. The method of claim 1 , wherein said second antigen comprises a first gG2Δ polypeptide, wherein said first gG2Δ polypeptide lacks the amino acid sequence of GHTNTSSAS (SEQ ID NO:07).
18. The method of claim 17 , wherein said first gG2Δ polypeptide comprises the amino acid sequence of SEQ ID NO:10.
19. The method of claim 17 , wherein said first gG2Δ polypeptide is selected from the group consisting of: gG2ΔJ24, gG2ΔJA, gG2ΔJA1, and gG2ΔJA2.
20. The method of claim 17 , wherein said first gG2Δ polypeptide comprises a substitution at the position corresponding to the amino acid sequence of SEQ ID NO:07, wherein said first gG2Δ polypeptide is selected from the group consisting of: gG2subJ24, gG2subJA, gG2subJA1, and gG2subJA2.
21. The method of claim 17 , wherein said first gG2Δ polypeptide comprises a substitution of about nine glycine residues at the position corresponding to the amino acid sequence of SEQ ID NO:07 in the native polypeptide.
22. The method of claim 1 , wherein said second antigen comprises a second gG2Δ polypeptide, wherein said second gG2Δ polypeptide lacks the amino acid sequence of AAKTPPTTPAP (SEQ ID NO:06).
23. The method of claim 22 , wherein said second gG2Δ polypeptide comprises the amino acid sequence of SEQ ID NO:12.
24. The method of claim 22 , wherein said second gG2Δ polypeptide is selected from the group consisting of: gG2ΔJ24, gG2ΔJA, and gG2ΔJA2.
25. The method of claim 22 , wherein said second gG2Δ polypeptide comprises a substitution at the position corresponding to the amino acid sequence of SEQ ID NO: 06, wherein said second gG2Δ polypeptide is selected from the group consisting of: gG2subJ24, gG2subJA, and gG2subJA2.
26. The method of claim 22 , wherein said second gG2Δ polypeptide comprises a substitution of about eleven glycine residues at the position corresponding to the amino acid sequence of SEQ ID NO:06 in the native polypeptide.
27. The method of claim 3 , wherein said third antigen is the amino acid sequence AAKTPPTTPAP (SEQ ID NO:06).
28. The method of claim 3 , wherein said third antigen comprises a polypeptide having an amino acid sequence comprising AAKTPPTTPAP (SEQ ID NO:06).
29. The method of claim 3 , wherein said second antigen is the full-length glycoprotein, gG2 and said third antigen comprises a polypeptide having an amino acid sequence comprising AAKTPPTTPAP (SEQ ID NO:06).
30. The method of claim 3 , wherein at least one of said antigens is immobilized on a nitrocellulose membrane support.
31. The method of claim 3 , wherein at least one of said antigens is immobilized on a solid support.
32. The method of claim 3 , wherein said solid support is selected from the group consisting of: a microparticle, an agarose bead, and a magnetic bead.
33. The method of claim 3 , wherein said first and said third antigens are not immobilized on a solid support and said second antigen is immobilized on a solid support.
34. The method of claim 3 , wherein at least one of said first and said third antigens is immobilized on a first solid support, and said second antigen is immobilized on a second solid support.
35. The method of claim 3 , wherein said antibodies bound to said first and said third antigens are removed from the sample prior to said detecting step.
36. The method of claim 3 , wherein said antibodies bound to said first and said third antigens are removed from the sample prior to contacting the sample with said second antigen.
37. The method of claim 3 , wherein at least one of said first and said third antigens is detectably labeled.
38. The method of claim 3 , wherein at least one of said first and said third antigens is a fusion protein.
39. The method of claim 4 , wherein said first antigen comprises a polypeptide having an amino acid sequence consisting of GHTNTSSAS (SEQ ID NO:07).