Immunity colloidal gold reagent for detecting IgM antibody of giant cells virus and its preparing method

A technology of cytomegalovirus and colloidal gold, which is applied in the field of preparation of the reagent, can solve problems such as the inability to realize single-person detection, the inability to realize one-step operation, and the long time required for detection, so as to achieve easy popularization and use, fast detection speed, low cost effect

Inactive Publication Date: 2003-07-23
博顿生物检验技术(杭州)有限公司
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  • Abstract
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  • Claims
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Problems solved by technology

The disadvantages of the enzyme-linked immunosorbent (EIA) method are: special instruments and equipment such as microplate readers are required for use; detection operators need to undergo professional training; the operation process is relatively complicated, and the time required for detection is relatively long; the cost of detection High, unable to achieve single-person detection
The disadvantages of the latex agglutination method...

Method used

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Examples

Experimental program
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Embodiment Construction

[0007] One end of the PVC backing is adhered sequentially to the sample pad, the binding pad coated with anti-u chain monoclonal antibody and colloidal gold conjugate, and the middle is adhered to the coated cytomegalovirus-specific surface membrane antigen and goat (or rabbit) antibody. Nitrocellulose membrane of mouse IgG with absorbent pad attached at the other end.

[0008] It was prepared according to the following steps: (1) Preparation of monoclonal antibody: the hybridoma cell line was routinely cultured and passaged in vitro, injected into the peritoneal cavity of BALB / C mice with human u-chain, collected ascites, and purified the antibody by affinity chromatography. U-chain monoclonal antibody; (2) Preparation of polyclonal antibody: immunize mice with human-derived u-chain multiple times, extract antiserum to immunize goats or rabbits, and obtain goat anti-mouse IgG or rabbit anti-mouse IgG after purification; (3 ) Preparation of colloidal gold: 100ml 0.01% gold chl...

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Abstract

The sample pad and the binding pad, which is the conjugate coated with the anti U chain monoclonal antibody and colloidal gold, are adhered to the one end with PVC back covered in sequence. The cellulose nitrate film coated with the specificity surface film antigen of giant cells virus and the anti rat IgG of sheep (or rabbit) is adhered to the middle of the reagent. The water uptake pad is adhered to another end of the reagent. The reagent features high specificity and sensivitiy, simple and fast. The reagent is applicable to the detection in site within 30 minutes for the full testing procedure. Operation personnel can complete the operation by reading the menu and without need of professional training.

Description

technical field [0001] The invention relates to an immune colloidal gold reagent for detecting antibodies, and also relates to a preparation method of the reagent. Background technique [0002] Existing methods for detecting CMV antibody IgM mainly include enzyme-linked immunosorbent assay, latex agglutination, immunocolloidal gold diafiltration and so on. The disadvantages of the enzyme-linked immunosorbent (EIA) method are: special instruments and equipment such as microplate readers are required for use; detection operators need to undergo professional training; the operation process is relatively complicated, and the time required for detection is relatively long; the cost of detection Higher, single-person detection cannot be realized. The disadvantages of the latex agglutination method are: the operation process is relatively complicated and demanding, and one-step operation cannot be realized; the reagent needs to be stored at low temperature; ...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/531G01N33/543G01N33/569
Inventor 张少恩
Owner 博顿生物检验技术(杭州)有限公司
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