Hepatitis C virus antigen-antibody joint inspection kit and application thereof

A hepatitis C virus, antigen antibody technology, applied in the field of immune detection, can solve the problem of detection sensitivity reduction, achieve the effect of improving detection sensitivity, reducing interference, and low detection cost

Pending Publication Date: 2020-08-11
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, once HCV-infected patients produce antibodies in vivo and undergo seroconversion, antigen-antibody complexes can be formed between anti-core antigen antibodies and HCVcAg, and the detection sensitivity will be significantly reduced

Method used

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  • Hepatitis C virus antigen-antibody joint inspection kit and application thereof
  • Hepatitis C virus antigen-antibody joint inspection kit and application thereof
  • Hepatitis C virus antigen-antibody joint inspection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0131] Example 1: Preparation of a kit for joint detection of hepatitis C virus antigen and antibody

[0132] 1. Preparation of receptor-HCV antibody 1 (against the N-terminal epitope of HCV core antigen)

[0133] 1) Antibody treatment: HCV antibody 1 is dialyzed, replaced with coating buffer, and the protein concentration is measured.

[0134] 2) Receptor treatment: Replace the receptor with coating buffer through centrifugation, ultrasound and other processes;

[0135] 3) Coupling: The processed receptor and the processed HCV antibody 1 are mixed and reacted, and the receptor-HCV antibody 1 is obtained after reduction and blocking processes, and the volume is constant and stored with the preservation solution.

[0136] 2. Preparation of biotin-HCV antibody 2 (for the C-terminal epitope of HCV core antigen)

[0137] 1) Antibody treatment: HCV antibody 2 is dialyzed, replaced with a labeling buffer, and the protein concentration is measured.

[0138] 2) Labeling reaction: the processed HC...

Embodiment 2

[0160] Example 2: Screening of the concentration of each component of the treatment agent in the kit

[0161] The detection results in the following examples are the results of detection using LITA HT.

[0162] The treatment agent in the following examples is used to process the sample to be tested in the antigen well in the HCV Ag / Ab combined test. The other components in the antigen well are FG-HCV-Ab1 and Bio-HCV-Ab2 (two strains are directed against the HCV core antigen For monoclonal antibodies with different epitopes, one monoclonal antibody binds to the receptor, and the other monoclonal antibody binds to biotin). There are two ways to use the treatment agent, namely:

[0163] Usage 1:

[0164] 1) Mix the treatment agent with the sample to be tested in a volume ratio of 1:1, and incubate at 37°C for 30 minutes to obtain the processed sample to be tested;

[0165] 2) Take 25ul of the processed sample to be tested and add it to the microplate, then add 25ul of FG-HCV-Ab1 solution...

Embodiment 3

[0205] Example 3: Effect of HCV antigen-antibody joint test method

[0206] 1. The main components of the kit used are as follows:

[0207] Treatment agent: 10% urea, 3% n-butanol, 1% Triton114, 0.75% CTAB and 9% sodium chloride;

[0208] Receptor-HCV antibody 1: The receptor-HCV antibody 1 reagent prepared in Example 1, and the receptor-HCV antibody is diluted 1 to 50ug / ml with a luminescent reagent buffer;

[0209] Biotin-HCV antibody 2: The biotin-HCV antibody 2 reagent prepared in Example 1, and the biotin-HCV antibody is diluted 2 to 1 ug / ml with biotin buffer;

[0210] Receptor-HCV antigen 1: The receptor-HCV antigen 1 reagent prepared in Example 1, and the receptor-HCV antigen 1 to 30ug / ml is diluted with a luminescent reagent buffer;

[0211] Biotin-HCV antigen 2: The biotin-HCV antigen 2 reagent prepared in Example 1, and the biotin-HCV antigen is diluted 2 to 0.5ug / ml with a biotin buffer;

[0212] Sample diluent: PB buffer containing sodium chloride, calf serum, etc.

[0213] 2....

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Abstract

The invention relates to a hepatitis C virus antigen-antibody joint detection kit and an application thereof, belongs to the technical field of immunodetection. The kit comprises the following components, a receptor bound with a first antigen, a second antigen, a receptor binding to the first antibody, and a second antibody, wherein the first antigen and the second antigen can be specifically combined with a variable region of a hepatitis C virus antibody, the first antibody and the second antibody can be specifically combined with different epitopes of a hepatitis C virus antigen. The kit isadvantaged in that the HCV antigen-antibody joint detection kit can be used for joint detection of HCV core antigen and antibody, so a detection window period is shortened, detection cost is low, moreover, the kit further comprises a treating agent, and the treating agent can reduce the interference of a low-affinity antibody in an early body and improve the antigen detection sensitivity in a conversion period, so the HCV detection sensitivity is improved, and the HCV detection window period is further shortened.

Description

Technical field [0001] The invention belongs to the technical field of immunoassays, and specifically relates to a kit for joint detection of hepatitis C virus antigen and antibody and its application. Background technique [0002] Hepatitis C virus (HCV) is a chronic hepatotropic virus. The clinical symptoms of patients infected with HCV are not obvious, and it is not easy to be found. The incidence of chronic HCV infection is relatively high, and cirrhosis and liver cancer are prone to occur earlier. At present, the commonly used hepatitis C virus (HCV) detection kit in China is an antibody detection kit. Although the sensitivity and specificity of HCV antibody detection technology are already high, there is still a period of time after HCV infection before the production of anti-HCV antibodies. The long period of about 40 to 70 days (66 days on average) is called the window period before seroconversion after infection. At this time, the HCV antibody detection kit cannot be de...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/543
CPCG01N33/5767G01N33/54313G01N33/56983
Inventor 吴晨赵琪张林钰李临
Owner CHEMCLIN DIAGNOSTICS CO LTD
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