Claims
- 1. A method for determining whether a subject has been vaccinated against hepatitis A virus or naturally infected with hepatitis A virus, said method comprising:
- (a) contacting a biological sample from said subject with HAV structural proteins under conditions which permit the formation of a complex between antibodies to the structural proteins and the structural proteins; and
- (b) contacting a sample which forms a complex in step (a) with HAV 3C proteinase under conditions which permit the formation of a complex between antibodies to the 3C proteinase and the 3C proteinase, wherein the detection of a complex between the antibodies to 3C proteinase and the 3C proteinase indicates that said subject has been naturally infected with hepatitis A virus and a failure to detect said complex indicates that the subject has been vaccinated against hepatitis A virus.
- 2. The method of claim 1, wherein the 3C proteinase in step (b) is bound to a solid support.
- 3. The method of claim 2, wherein the biological sample is contacted in step (b) with a recombinantly expressed 3C proteinase.
- 4. The method of claim 3, wherein the antibody-proteinase complex in step (b) is detected using a labelled secondary antibody.
- 5. The method of claim 4, wherein the antibodies detected in step (b) are IgG antibodies.
- 6. The method of claim 4, wherein the antibodies detected in step (b) are IgM antibodies.
- 7. The method of claim 1, wherein the presence of antibodies to 3C proteinase is detected in step (b) by measuring the 3C proteinase activity of the 3C proteinase contacted with the biological sample, an inhibition of 3C proteinase activity as compared to the 3C proteinase activity of 3C proteinase not contacted with the biological sample indicating the presence of antibodies to 3C proteinase in the biological sample.
- 8. The method of claim 7, wherein the 3C proteinase activity is determined by incubating the 3C proteinase with at least one 3C proteinase substrate.
- 9. The method of claim 8, wherein the 3C proteinase substrate is labelled.
- 10. A method for determining whether a subject has been infected with hepatitis A virus; said method comprising contacting a biological sample from said subject with HAV 3C proteinase under conditions which permit the formation of a complex between antibodies to the 3C proteinase and the 3C proteinase, wherein the detection of a complex between the antibodies to 3C proteinase and the 3C proteinase indicates that said subject has been naturally infected with HAV.
- 11. The method of claim 10, wherein the 3C proteinase is bound to a solid support.
- 12. The method of claim 11, wherein the biological sample is contacted with a recombinantly expressed 3C proteinase.
- 13. The method of claim 12, wherein the antibody-proteinase complex is detected using a labelled secondary antibody.
- 14. The method of claim 13, wherein the antibodies detected are IgG antibodies.
- 15. The method of claim 13, wherein the antibodies detected are IgM antibodies.
- 16. The method of claim 10, wherein the presence of antibodies to 3C proteinase is detected by measuring the 3C proteinase activity of the 3C proteinase contacted with the biological sample, an inhibition of 3C proteinase activity as compared to the 3C proteinase activity of 3C proteinase not contacted with the biological sample indicating the presence of antibodies to 3C proteinase in the biological sample.
- 17. The method of claim wherein the 3C proteinase activity is determined by incubating the 3C proteinase with at least one 3C proteinase substrate.
- 18. The method of claim 17, wherein the 3C proteinase substrate is labelled.
- 19. A method for determining whether a subject seropositive for antibodies to hepatitis A virus structural proteins is seropositive as a result of infection with hepatitis A virus or of vaccination against hepatitis A virus, said method comprising contacting a biological sample from said patient with HAV 3C proteinase under conditions which permit the formation of a complex between antibodies to 3C proteinase and the 3C proteinase, wherein the detection of a complex between the antibodies to 3C proteinase and the 3C proteinase indicates that said subject has been naturally infected with hepatitis A virus and a failure to detect said complex indicates that the individual has been vaccinated against hepatitis A virus.
Parent Case Info
This application is a 371 of PCT/US97/03428, Filed Mar. 13, 1997. This application claims benefit of provisional application 60/013,333, Filed Mar. 13, 1996.
PCT Information
Filing Document |
Filing Date |
Country |
Kind |
102e Date |
371c Date |
PCT/US97/03428 |
3/13/1997 |
|
|
9/3/1998 |
9/3/1998 |
Publishing Document |
Publishing Date |
Country |
Kind |
WO97/34136 |
9/18/1997 |
|
|
Non-Patent Literature Citations (3)
Entry |
Robertson et al., Antibody Response to Nonstructural Proteins of Hepatitis A Virus Following Infection. Journal of Medical Virology 40:76-82, 1993. |
Schultheiss et al., Clevage Specificity of Purified Recombinant Hepatitis A Virus 3C Proteinase on Natural Substrates, Journal of Virology 69(3):1727-1733, 1995. |
Jokik et al., Eds, Zinsser Microbiology, 20th Ed., 1992 Appleton and Lange, Norwalk, p. 1040. |