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 specific anti-herpes simplex virus type-2 (HSV-2) antibodies in a biological sample, comprising:
(a) preabsorbing a biological sample suspected of containing anti-HSV-2 antibodies by contacting the sample with a first preabsorbing antigen comprising the amino acid sequence AAKTPPTTPAP (SEQ ID NO:06), wherein said first preabsorbing antigen is not a full length glycoprotein G2 (gG2) polypeptide and does not deplete said specific anti-HSV-2 antibodies,
(b) contacting said biological sample with a second antigen capable of binding a 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.
2. The method of claim 1 , wherein said first preabsorbing antigen comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:3, and SEQ ID NO:2.
3. The method of claim 2 , wherein said first preabsorbing antigen consists of a detectable label and an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:3, and SEQ ID NO:2.
4. The method of claim 1 , wherein said detecting further comprises contacting said sample with one or more detectably labeled anti-human immunoglobulin antibodies.
5. The method of claim 1 , wherein at least one of said antigens is immobilized on a nitrocellulose membrane support.
6. The method of claim 1 , wherein at least one of said antigens is immobilized on a solid support.
7. The method of claim 6 , wherein said solid support is selected from the group consisting of: a microparticle, an agarose bead, and a magnetic bead.
8. The method of claim 1 , wherein said first preabsorbing antigen is not immobilized on a solid support and said second antigen is immobilized on a solid support.
9. The method of claim 1 , wherein said first preabsorbing antigen is immobilized on a first solid support, and said second antigen is immobilized on a second solid support.
10. The method of claim 1 , wherein said antibodies bound to said first preabsorbing antigen are removed from the sample prior to step (c).
11. The method of claim 1 , wherein said antibodies bound to said first preabsorbing antigen are removed from the sample prior to step (b).
12. The method of claim 1 , wherein said first preabsorbing antigen is detectably labeled.
13. 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.
14. The method of claim 1 , wherein said second antigen comprises an epitope from the glycoprotein, gG2.
15. The method of claim 1 , wherein said second antigen is the full-length glycoprotein gG2.
16. 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 AAKTPPTTPAP (SEQ ID NO:06).
17. The method of claim 16 , wherein said first gG2Δ polypeptide comprises the amino acid sequence of SEQ ID NO:12.
18. The method of claim 16 , wherein said first gG2Δ polypeptide is selected from the group consisting of: gG2ΔJ24, gG2ΔJA, and gG2ΔJA2.
19. The method of claim 16 , wherein said first gG2Δ polypeptide comprises a substitution at the position corresponding to the amino acid sequence of SEQ ID NO: 06, wherein said first gG2Δ polypeptide is selected from the group consisting of: gG2subJ24, gG2subJA, and gG2subJA2.
20. The method of claim 16 , wherein said first 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.
21. 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 GHTNTSSAS (SEQ ID NO:07).
22. The method of claim 21 , wherein said second gG2Δ polypeptide comprises the amino acid sequence of SEQ ID NO:10.
23. The method of claim 21 , wherein said second gG2Δ polypeptide is selected from the group consisting of: gG2ΔJ24, gG2ΔJA, gG2ΔJA1, and gG2ΔJA2.
24. The method of claim 21 , wherein said second gG2Δ polypeptide comprises a substitution at the position corresponding to the amino acid sequence of SEQ ID NO:07, wherein said second gG2Δ polypeptide is selected from the group consisting of: gG2subJ24, gG2subJA, gG2subJA1, and gG2subJA2.
25. The method of claim 21 , wherein said second 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.