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Combined detection kit for hepatitis c virus (HCV) antigen-antibody

A hepatitis C virus, antigen-antibody technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of inability to detect HCV antibodies, missed detection, and missed antibody detection.

Inactive Publication Date: 2016-11-09
HUNAN KANGRUN PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, because patients will produce antibodies after being infected with hepatitis C virus for about a period of time (average 49 days), and form antigen-antibody immune complexes. Yield is greatly reduced
Generally, detergents and denaturants are added to the sample diluent for the detection of HCV antigens to destroy the complex and release the antigens. However, such detergents and denaturants also destroy the HCV antibodies in the samples, making it impossible to detect HCV in the joint test reagents. Antibodies, resulting in missed detection of antibodies
[0007] The combined detection kits for hepatitis C virus antigen and antibody currently on the market use mild detergents and denaturants as much as possible to minimize the impact on the HCV antibody in the sample to be tested. However, if the strength of the detergents and denaturants is not strong or If the concentration is not enough, the immune complex in the sample to be tested will not be completely destroyed, the HCV core antigen cannot be completely released, the content of HCV core antigen in the sample to be tested will be small, and the detection sensitivity for HCV core antigen and HCV antibody is low , it is easy to cause missed detection

Method used

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  • Combined detection kit for hepatitis c virus (HCV) antigen-antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] The first sample dilution and the second sample dilution were prepared with phosphate buffer, and the specific components were as follows.

[0077] First sample dilution: 0.5 wt% β-mercaptoethanol, 0.05 wt% sodium dodecylsulfonate, 0.03 wt% Tween-20.

[0078] Second sample diluent: 3wt% sodium lauryl sarcosine and 3mol / L urea.

[0079] Dilute AB1 with carbonate buffer to a concentration of 3 μg / mL, dilute AG1 with carbonate buffer to a concentration of 6 μg / mL, and mix the two at a volume ratio of 1:1 to obtain a coating solution.

[0080] Then add the coating solution into the polystyrene microwell plate, and complete the coating after adsorption, washing, sealing and drying.

[0081] Add 100 μL of sample / positive control / negative control and 50 μL of the first sample dilution to each well, incubate at 37°C for 30 minutes without washing the plate, then add 50 μL of the second sample dilution to each well, incubate at 37°C for 30 minutes and wash the plate 6 times.

...

Embodiment 2

[0085] The first sample dilution and the second sample dilution were prepared with phosphate buffer, and the specific components were as follows.

[0086] The first sample dilution: 0.3wt% dithiothreitol, 0.01wt% sodium dodecylsulfonate, 0.01wt% guanidine hydrochloride.

[0087] The second sample diluent: 1 wt% dodecyl dimethyl betaine and 5 mol / L urea.

[0088] Dilute AB1 with carbonate buffer to a concentration of 3 μg / mL, dilute AG1 with carbonate buffer to a concentration of 6 μg / mL, and mix the two at a volume ratio of 1:1 to obtain a coating solution.

[0089] Then add the coating solution into the polystyrene microwell plate, and complete the coating after adsorption, washing, sealing and drying.

[0090] Add 100 μL of sample / positive control / negative control and 50 μL of the first sample dilution to each well, incubate at 37°C for 30 minutes without washing the plate, then add 50 μL of the second sample dilution to each well, incubate at 37°C for 30 minutes and wash t...

Embodiment 3

[0094] The first sample dilution and the second sample dilution were prepared with phosphate buffer, and the specific components were as follows.

[0095] First sample dilution: 1 wt% cysteine, 0.08 wt% sodium dodecylsulfonate, 0.1 wt% guanidine hydrochloride.

[0096] The second sample diluent: 5 wt% dodecyl dimethyl betaine and 1 mol / L polyethylene glycol octyl phenyl ether.

[0097] Dilute AB1 with carbonate buffer to a concentration of 3 μg / mL, dilute AG1 with carbonate buffer to a concentration of 6 μg / mL, and mix the two at a volume ratio of 1:1 to obtain a coating solution.

[0098] Then add the coating solution into the polystyrene microwell plate, and complete the coating after adsorption, washing, sealing and drying.

[0099] Add 100 μL of sample / positive control / negative control and 50 μL of the first sample dilution to each well, incubate at 37°C for 30 minutes without washing the plate, then add 50 μL of the second sample dilution to each well, incubate at 37°C f...

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Abstract

The invention discloses a combined detection kit for HCV antigen-antibody. The kit comprises a first monoclonal antibody against HCV core, a first HCV recombinant chimeric antigen, a first diluted sample solution, a second diluted sample solution, a first enzyme-labeled second monoclonal antibody against HCV core antigen, a first ligand-labeled second HCV recombinant chimeric antigen, a second enzyme-labeled second ligand, a developer and a stopping solution, wherein the first diluted sample solution comprises a reducing agent, a first surfactant and a first denaturant; and the second diluted sample solution comprises a second surfactant and a second denaturant. The combined detection kit for the HCV antigen-antibody opens mismatched disulfide bonds in coating antigen and in an antigen-antibody complex of a sample in virtue of the first diluted sample solution, and destroys HCV core antigen and HCV antibody compounds in virtue of the second diluted sample solution, so more antigen is released; and thus, detection sensitivity is improved.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a hepatitis C virus antigen antibody combined detection kit. Background technique [0002] Hepatitis C is a kind of acute and chronic inflammation of the liver caused by hepatitis C virus (Hepatitis C virus, HCV). According to WHO statistics, there are currently about 200 million people infected with hepatitis C virus in the world, accounting for about 30% of the global population. %, there are about 40 million to 60 million in China. Hepatitis C is more prone to liver cirrhosis, fibrosis, and liver cancer than hepatitis B. It is predicted that in the near future, hepatitis C will replace hepatitis B and become another major virus killer that threatens human health after HIV. Although hepatitis C virus is very harmful to human health, there is no vaccine to prevent it. Early diagnosis of HCV and screening of infectious sources can guide clinical treatment and disease control....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/576
CPCG01N33/5767
Inventor 胡道奇毛金武周咏武李光
Owner HUNAN KANGRUN PHARMA
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