Immunoassay method of HIV-1p24 antigen and reagent therefor

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage

Reexamination Certificate

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C435S007100, C435S007930, C435S007940, C424S130100, C424S160100, C424S208100

Reexamination Certificate

active

06432633

ABSTRACT:

BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a sandwich immunoassay method of the HIV-1p24 antigen and a reagent for use in the sandwich immunoassay method.
2. Description of the Related Art
Acquired immunodeficiency syndrome (AIDS) means a group of diseases triggered by human immunodeficiency virus (HIV). Since HIV infection via blood was revealed, transfusion blood samples have been subjected to HIV antibody screening at blood centers all over Japan. Because the antibody titer never increases 6 to 8 weeks after HIV infection, however, the current HIV antibody screening methods are problematic in that the window-period (blank period) is present, in which blood infected with HIV cannot be detected although the blood is HIV positive.
Besides HIV antibody screening, alternatively, attempts have been made about the shortening of the window-period by screening blood for the presence of HIV antigen. HIV antigen is appropriately assayed by immunoassay methods for screening a great number of samples, but the antigenicity of HIV antigen is readily exposed to mutagenesis. Therefore, attempts have been made to assay the HIV-1p24 antigen with relatively stable antigenicity, in particular. HIV Antigen·EIA “ABBOTT” is commercially available as a product for HIV-1p24 antigen assay. So as to avoid the error in the detection of infected blood as much as possible, an assay method rapidly assaying a great number of samples at a higher detection sensitivity is desired.
It is a purpose of the invention to provide an immunoassay method of HIV-1p24 antigen at a high detection sensitivity and a reagent therefor.
SUMMARY OF THE INVENTION
So as to overcome such conventional problems, the present inventors have made investigations about the assay of HIV-1p24 antigen. The inventors have successfully assayed the HIV-1p24 antigen by a sandwich immunoassay method using a polyclonal antibody recognizing the C-terminal region of the HIV-1p24 antigen and two monoclonal antibodies recognizing the epitopes except the C-terminal region of the HIV-1p24 antigen, at a far higher detection sensitivity than those by the existing assay methods of the HIV-1p24 antigen. Thus, the invention has been achieved.
More specifically, the invention provides a sandwich immunoassay method of HIV-1p24 antigen, using a polyclonal antibody recognizing the C-terminal region of the HIV-1p24 antigen and at least two monoclonal antibodies recognizing the HIV-1p24 antigen and further, the HIV-1p24 antigen and the HIV-2p25 antigen, along with a reagent kit for use in the immunoassay method.
By using the inventive immunoassay method, HIV-1p24 antigen and HIV-2p25 antigen can be assayed at high detection sensitivity. Thus, the inventive immunoassay method can be widely used for the immunoassay of the HIV-1p24 antigen and HIV-2p25 antigen and the screening of transfusion blood.


REFERENCES:
patent: 6107049 (2000-08-01), Allard et al.
patent: WO 91/13360 (1991-09-01), None
Schupbach et al., AIDS, vol. 10 (1996) pp. 1085-1090.
Jackson et al., J. of A.I.D.S., vol. 2 (1989) pp. 394-497.
Willoughby et al., Diagn. Microbiol. Infect. Ds., vol. 12 (1989) pp. 319-326.

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