IgA (Immunoglobulin A) antibody detection reagent kit (colloidal gold method) for EB (Epstein-Barr) viruses and preparation method thereof

A technology for antibody detection and Epstein-Barr virus, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of complex operation and long detection time of ELISA method

Inactive Publication Date: 2011-01-19
北京中检安泰诊断科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the ELISA method is complicated to operate and takes a long time to detect. It needs to perform operations such as dilution, incubation, separation, washing and color development of the sample to be tested, which takes more than two hours.

Method used

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  • IgA (Immunoglobulin A) antibody detection reagent kit (colloidal gold method) for EB (Epstein-Barr) viruses and preparation method thereof
  • IgA (Immunoglobulin A) antibody detection reagent kit (colloidal gold method) for EB (Epstein-Barr) viruses and preparation method thereof
  • IgA (Immunoglobulin A) antibody detection reagent kit (colloidal gold method) for EB (Epstein-Barr) viruses and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Such as figure 1 As shown, the production process of the EB virus IgA antibody detection kit (colloidal gold method) finished product is carried out according to the following steps.

[0084] 1. Raw material requirements:

[0085] 1.1 Recombinant Epstein-Barr virus antigen

[0086] (1) Appearance: colorless transparent liquid;

[0087] (2) Concentration and purity requirements: the concentration is greater than 2 mg / ml, determined by SDS-PAGE, and only one band exists under the condition of 10 μl of sample loading;

[0088] 1.2 Goat anti-mouse IgG antibody

[0089] (1) Appearance: colorless transparent liquid;

[0090] (2) Concentration requirements: concentration greater than 4mg / ml;

[0091] 1.3 Mouse anti-human IgA monoclonal antibody

[0092] (1) Appearance: colorless transparent liquid;

[0093] (2) Concentration and purity requirements: the concentration is greater than 2 mg / ml, determined by SDS-PAGE, and only one band exists under the condition of 10 μl o...

Embodiment 2

[0182] Kit preparation one

[0183] (1) Preparation of reaction membrane: Dilute the recombinant Epstein-Barr virus antigen to a coating concentration of 1.0 mg / ml with phosphate buffer, dilute the goat anti-mouse IgG antibody to a coating concentration of 2.0 mg / ml, and use a film dispenser Draw the two coating solutions onto the nitrocellulose membrane respectively, dry the coated nitrocellulose membrane and store it;

[0184] (2) Preparation of mouse anti-human IgA monoclonal antibody gold conjugate pad: Colloidal gold-labeled mouse anti-human IgA monoclonal antibody solution was obtained by coupling mouse anti-human IgA monoclonal antibody with a labeling concentration of 10 μg / ml to colloidal gold. Centrifuge at 12000r / min for 40min, discard the supernatant, add the colloidal gold conjugate diluent to 1 / 2 of the original volume, then soak the gold standard pad with 1.5ml / strip of the mixed solution to prepare EB-IgA gold conjugate pad , put the EB-IgA gold conjugate pad...

Embodiment 3

[0188] Kit Preparation II

[0189] (1) Preparation of reaction membrane: Dilute the recombinant Epstein-Barr virus antigen to a coating concentration of 1.0 mg / ml with phosphate buffer, dilute the goat anti-mouse IgG antibody to a coating concentration of 2.0 mg / ml, and use a film dispenser Draw the two coating solutions onto the nitrocellulose membrane respectively, dry the coated nitrocellulose membrane and store it;

[0190] (2) Preparation of mouse anti-human IgA monoclonal antibody gold conjugate pad: Colloidal gold-labeled mouse anti-human IgA monoclonal antibody solution was obtained by coupling mouse anti-human IgA monoclonal antibody with a labeling concentration of 23 μg / ml to colloidal gold. Centrifuge at 12000r / min for 40min, discard the supernatant, add the colloidal gold conjugate diluent to 1 / 2 of the original volume, then soak the gold standard pad with 1.5ml / strip of the mixed solution to prepare EB-IgA gold conjugate pad , put the EB-IgA gold conjugate pad in...

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Abstract

The invention relates to an IgA (Immunoglobulin A) antibody detection reagent kit (a colloidal gold method) for EB (Epstein-Barr) viruses and a preparation method thereof. The reagent kit comprises recombination antigen EB-NA1 coated by a nitrocellulose membrane detection line, a goat-anti-mouse IgG antibody coated on a quality control line and a mouse-anti-human IGA monoclonal antibody marked by colloidal gold and coated on a gold mark pad. The preparation method comprises the steps of: preparing a reaction membrane and a mouse-anti-human IGA monoclonal antibody gold combo pad, cutting and assembling to prepare the product. The invention has the advantages that: the IgA antibody detection reagent kit for the EB viruses has the characteristics of fast, simple and convenient detection, and high accuracy and sensitivity; the integrated operation time only requires 20 minutes to judge and read results; the colloidal gold is used for fast detecting test paper; a multi-epitope recombination antigen is used as a raw material; the method has the characteristics of simple and convenient operation, low cost, good specificity, high sensitivity, single portion detection and easy popularization; and the detection and control effect to the EB viruses is obvious.

Description

technical field [0001] The invention relates to an antibody detection kit, in particular to a kit and a preparation method for detecting Epstein-Barr virus IgA antibody by colloidal gold method. Background technique [0002] Epstein-Barr virus (EBV) is the only lymphoid follicular virus that can cause human infection in the Herpesviridae Gamma subfamily. It was first discovered in 1964 by Epstein and Barr from the cell culture of malignant lymphoma in African children, so it was named Epstein-Barr virus is the earliest recognized human tumor virus. [0003] Enzyme-linked immunosorbent assay (ELISA) is the most commonly used method to detect EB antibody in serum / plasma. Commercial kits are available, such as ELISA and indirect immunoenzyme staining (IEA) to detect IgA2VCA antibody in nasopharyngeal carcinoma serum 1. For the antibody titer of EBV 2EA early complex antigen (EBV 2EA) IgG, under the premise of the same sensitivity (88.3%), the specificity (98.3%) of ELISA metho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569G01N33/531
Inventor 周晟
Owner 北京中检安泰诊断科技有限公司
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