A homogeneous immunoassay method and its application

An immunoassay method and detection area technology, which is applied in the field of homogeneous immunoassay, can solve the problem of not being able to distinguish whether a positive result is antibody positive or antigen positive, and achieve the effect of low detection sensitivity

Active Publication Date: 2022-05-13
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide an efficient and reliable homogeneous immunoassay method for combined detection of antigen and antibody in view of the deficiencies of the prior art. The existence of complexes of antigen and / or antibody to be tested solves the problem that in the homogeneous immunoassay method, the combined detection of antigen and antibody cannot distinguish whether the positive result is antibody positive or antigen positive

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  • A homogeneous immunoassay method and its application
  • A homogeneous immunoassay method and its application
  • A homogeneous immunoassay method and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0143] Example 1: Double-hole combined detection of antigen and antibody in HIV clinical samples to be tested by homogeneous immunization method

[0144] The main reagents used include:

[0145] sample diluent;

[0146] A mixed solution of HIV gp41-type antigen-binding receptors and HIV gp36-type antigen-binding receptors;

[0147] A mixed solution of biotin-labeled HIV gp41 antigen and biotin-labeled HIV gp36 antigen;

[0148] Receptor solution combined with monoclonal antibody against HIV p24 antigen;

[0149] Biotin-labeled anti-HIV p24 polyclonal antibody solution;

[0150] Streptavidin-labeled donor solution.

[0151] Wherein the acceptor used in the present invention is prepared according to the example described in the patent PCT / US2010 / 025433, and its structure after being connected with HIV antigen / antibody is: HIV antigen / antibody-BSA-(dimethylthiophene) -(BHHCT), thereby obtaining receptors combined with HIV gp41 type antigen, receptors combined with HIV gp36 t...

Embodiment 2

[0164] Example 2: Double-hole combined detection of antigen and antibody in HIV clinical samples to be tested by homogeneous immunization method

[0165] The main reagents adopted in the following examples include:

[0166] sample diluent;

[0167] A mixed solution of receptor microspheres bound to HIV gp41 type antigen and receptor microspheres bound to HIV gp36 type antigen;

[0168] A mixed solution of biotin-labeled HIV gp41 antigen and biotin-labeled HIV gp36 antigen;

[0169] Receptor microsphere solution combined with monoclonal antibody against HIV p24 antigen;

[0170] Biotin-labeled anti-HIV p24 polyclonal antibody solution;

[0171] Streptavidin-labeled donor microsphere solution.

[0172] Wherein as the preparation method, composition structure and content of acceptor microspheres of the present invention can refer to the embodiment 1 of Chinese patent CN100429197C, then HIV antigen / antibody is coated on the surface of acceptor microspheres, to form HIV gp41 T...

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Abstract

The present invention relates to a homogeneous immunoassay method, which comprises at least two detection areas, in one detection area it is judged whether there is a first complex formed by donor-to-be-detected antibody-acceptor, and in another detection area Judging whether there is a second complex formed by donor-antigen-receptor; wherein, the donor can generate singlet oxygen in an excited state, and the acceptor can react with singlet oxygen to produce detectable signal of. The method solves the problem of HOOK effect of high-concentration samples and low detection sensitivity of low-concentration samples in the one-step detection of antigen and antibody. At the same time, the method also solves the problem that the positive result cannot be distinguished from antibody positive or antigen positive in combined detection of antigen and antibody. The method of the present invention is especially suitable for combined detection of HIV antigen and antibody.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a homogeneous immunoassay method and application thereof. Background technique [0002] Immunoassay method is a highly selective biochemical method established by the specific combination of antigen and antibody. Immunoassays are divided into labeled immunoassays and non-labeled immunoassays according to whether they are labeled or not. Due to the introduction of markers, the separation of the bound antigen-antibody from the excess antigen or antibody is inevitable, so the marker immunoassay is divided into homogeneous immunoassay and heterogeneous immunoassay according to whether it is separated or not. . The heterogeneous immunoassay method requires multi-step operations such as embedding, elution, and separation. The analysis process is cumbersome and the analysis time is long, which cannot meet the requirements of rapid detection and diagnosis. The homogeneous meth...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/569
Inventor 王志新刘宇卉李临
Owner CHEMCLIN DIAGNOSTICS CO LTD
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