Method for simultaneously detecting an antigen of, and an antibody against, an infectious microorganism
AssigneeBio-Rad Pasteur
InventorsFrançois Rieunier, Muriel Feyssaguet, Stéphanie Henriot, Nadine Lambert
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Abstract
The invention relates to a method for detecting, in vitro, an infection with a microorganism, such as the hepatitis C virus, in a biological sample, by simultaneously detecting an antigen of this microorganism and the antibodies against this same antigen, and also to the reagents and kits implementing this method.
Claims10
A method for detecting, in vitro, a hepatitis C virus (HCV) in a biological sample, comprising simultaneous detection of when present in the biological sample, at least one HCV antigen and an anti-HCV antibody, the method comprising
- a) contacting the biological sample with a HCV capture antibody and a HCV capture antigen attached to a solid phase;
- b) incubating the mixture under conditions allowing formation of antigen-antibody complexes;
- c) separating the solid phase from the liquid phase;
- d) revealing the antigen-antibody complexes formed by contacting the solid phase with, firstly, a labeled detection antibody capable of binding the HCV antigen which has been captured and, secondly, a labeled anti-immunoglobulin or anti-isotype antibody, or a labeled detection antigen, capable of binding the anti-HCV antibody which has been captured;
- each of the antigen for capturing and the antigen for detecting anti-HCV antibodies comprising an antigenic fragment of the capsid protein of HCV, in which two epitopes have been destroyed,
- the capture and detection antibodies each recognizing one of said epitopes, which are intact, of the capsid antigen which has been captured, and
- in which the capture antibody and the detection antibody each recognize an epitope selected from the group of epitopes having the following sequences:
- 44 LGVR47 (SEQ ID NO:
- 19), 30IVGGVYL36 (SEQ ID NO:
- 20), 29QIVGGV34 (SEQ ID NO:
- 21).
A kit which is of use for detecting an infection with a hepatitis C virus (HCV) in a biological sample, comprising:
- a capture or detection antigen which comprises an antigenic fragment of a protein of HCV, said fragment comprising at least one destroyed epitope, while at the same time conserving the ability to bind the antibodies directed against HCV-possibly present in the biological sample and being selected from the group consisting of SEQ ID NO:
- 4-18;
- a capture or detection antibody directed against said epitope, which is intact, of said protein of HCV.
A method for detecting in vitro, an infection with hepatitis C virus (HCV) in a biological sample, comprising simultaneous detection of at least one antigen of said HCV and of an antibody directed against said HCV, present in the biological sample, which method comprises
- a) bringing the biological sample into contact with a capture antibody directed against said HCV, and a capture antigen derived from said HCV;
- b) incubating the mixture under conditions allowing formation of antigen-antibody complexes;
- c) revealing the antigen-antibody complexes formed which optionally uses a labeled detection antibody capable of binding to the antigen of said HCV which has been captured and/or optionally also a labeled detection antigen capable of binding to the antibody directed against said HCV which has been captured;
- wherein the capture antigen and/or the labeled detection antigen of said HCV comprises an antigenic fragment of the capsid protein of HCV in which at least one epitope has been destroyed;
- wherein the capture and/or detection antibody recognizes said at least one epitope, which is intact, of the antigen which has been captured;
- d) the presence of capture antibody/antigen complexes and of antibody/capture antigen complexes indicating an infection with HCV,
- in which the capture antigen or the detection antigen is selected from the group consisting of SEQ ID NOS:
- 4-18.
Dependencies are inferred from the claim text.