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Enzyme-linked immunologic diagnosis kit for core antigen of C type hepatitis virus and method for preparing same

A technology of hepatitis C virus and diagnostic kits, applied in biological testing, material inspection products, measuring devices, etc., can solve the problems of not applying for invention patents in China, high prices, promotion and popularization, etc., achieve good application prospects, shorten Window period, effect of improving sensitivity

Inactive Publication Date: 2006-12-13
湖南景达基因有限公司
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Problems solved by technology

Due to technical confidentiality and other reasons, ORTHO has not applied for registration of new products in China, nor has it applied for invention patents in China. At the same time, due to the high price, the product has not been promoted and popularized in China.

Method used

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  • Enzyme-linked immunologic diagnosis kit for core antigen of C type hepatitis virus and method for preparing same
  • Enzyme-linked immunologic diagnosis kit for core antigen of C type hepatitis virus and method for preparing same
  • Enzyme-linked immunologic diagnosis kit for core antigen of C type hepatitis virus and method for preparing same

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Experimental program
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Embodiment Construction

[0029] 1. Preparation of HCV-core recombinant antigen

[0030] 1.1 Recombinant antigen cloning and expression: call out the full sequence gene of HCV from the gene bank, and analyze the sequences of the core antigens of different subtypes of HCV:

[0031]

[0032] At the same time, the Macvector program was used to analyze the antigenicity, hydrophilicity and antigenic epitope of the hepatitis C core antigen 1-160 amino acid sequence ( figure 2 ). Among them, there are 4 antigenic epitopes with conserved sequences and strong antigenicity, which are located at amino acid residues 20-35, 35-50 and 100-115, 115-130, respectively. The 2-160 amino acid sequence was selected and inserted into the expression vector pBVIL1 for recombinant cloning and expression ( image 3 ).

[0033] 1.2 Purification and activity identification of recombinant HCV-cAg: The recombinant HCV-C antigen extracted from engineering bacteria was eluted by S-Sepharose column step gradient, and the elutio...

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Abstract

Disclosed are a hepatitis C virus antigen enzyme-linked immunoassay reagent box and a method for making the same. The invention obtains the cell strain of excretive anti HCV core antigen by analyzing the core antigen array of the hepatitis C virus different type and cloning the core antigen gene of HCV, purifying out the high activity monoclonal antibody with four core aa expression sites of HCV, wherein the Cab1 and Cabs are used as coating antibodies, the Cab3 and Cab4 are used as enzyme labeled antibodies; employing double antibodies sandwich technology to prepare HCV-cAg ELISA diagnosing reagent box.

Description

technical field [0001] The invention relates to a virus antigen detection kit and a preparation method thereof, in particular to a hepatitis C virus core antigen (HCV-core Ag) ELISA diagnostic kit and a preparation method of the kit. Background technique [0002] Hepatitis C is caused by Hepatitis C Virus (HCV) infection and is mainly non-A, non-B hepatitis transmitted through blood transfusion. According to statistics, there are about 170 million HCV infected people in the world, and the number of HCV infected people in my country has reached 40 million. . At present, there is no effective clinical treatment method, and the vaccine against HCV has not yet been released. This situation has attracted great attention from the relevant departments of the Chinese government, especially in the field of blood product production. The country has promulgated relevant regulations to strictly control its quality. . However, due to the long "window period" of the current HCV Ab-ELISA ...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/544G01N33/52G01N33/535
Inventor 李少波张贺秋刘劭钢王国华李宏实陈坤周祖岳李凯周松华
Owner 湖南景达基因有限公司
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