Colloidal gold immunochromatographic test strip for detecting in-vivo neutralizing antibody after injection of novel coronavirus vaccine and preparation method of colloidal gold immunochromatographic test strip

A technology of immunochromatography test paper and colloidal gold, which is applied in the direction of immunoassay, biological test, measuring device, etc., to achieve the effects of lower detection limit, stable combination, and small batch-to-batch difference

Pending Publication Date: 2021-03-12
GUILIN UNIV OF ELECTRONIC TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] At present, there is no specific drug for COVID-19, and the n

Method used

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  • Colloidal gold immunochromatographic test strip for detecting in-vivo neutralizing antibody after injection of novel coronavirus vaccine and preparation method of colloidal gold immunochromatographic test strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1: the preparation of colloidal gold immunochromatography test strip

[0057] The colloidal gold immunochromatographic test strip is prepared according to the following method, and its structure is as follows: figure 1 Shown:

[0058] Paste the sample pad 1, the colloidal gold pad 2, the nitrocellulose membrane 3 and the absorbent filter paper 6 on the polyvinyl chloride (PVC) base plate 7 from left to right, and then place the PVC base plate 7 and the attached materials Cut into test strips with a width of 3mm, and put them into a card case, which is provided with a sample loading area and a color developing area (observation area).

[0059] Among them, sample pad 1 is a glass fiber membrane; colloidal gold pad 2 is coated with colloidal gold-labeled recombinant new coronavirus RBD protein; nitrocellulose membrane 8 is coated with angiotensin converting enzyme 2 (ACE2) detection line 4 , also coated with mouse anti-chicken IgG antibody as quality control l...

Embodiment 2

[0060] Example 2: Antibody coating

[0061] First, dilute the mouse anti-chicken IgG antibody and angiotensin-converting enzyme 2 (ACE2) with coating buffer 0.05mol / L PB with a pH of 7.2-7.4 to obtain the detection line 4 mouse anti-chicken IgG antibody coating solution and quality control Line 5 Angiotensin Converting Enzyme 2 (ACE2) Diluent;

[0062] Secondly, draw the obtained dilution onto the nitrocellulose membrane using a gold-spray film marking device, and draw the line at a condition of 0.1 μL / cm;

[0063] Finally, the nitrocellulose membrane was dried in an oven at 37°C for 3 hours, and sealed with a desiccant after drying;

Embodiment 3

[0064] Embodiment 3: the preparation of colloidal gold pad

[0065] (1) Wash the utensils, adjust the pH value of the colloidal gold particles to 8.5 with potassium nitrate, add them to the utensils, then add PEG6000 separating agent with a final concentration of 10%, and then add the recombinant novel coronavirus RBD protein to a final concentration of 30 μg / ml, stirred for 40min to obtain a colloidal gold solution;

[0066] (2) Add 10% BSA blocking solution to the colloidal gold solution obtained in step (1) to a final concentration of 1% for blocking for 15 minutes;

[0067] (3) Centrifuge the colloidal gold solution obtained in step (2) at 4°C and 12,000 rpm for 40 minutes, remove the supernatant after centrifugation, and redissolve the precipitate with a reconstitution solution to obtain a recombinant novel coronavirus RBD protein labeling solution;

[0068] The complex solution is a mixture of Tween 20 and BSA;

[0069] (4) Spray the recombinant 2019-nCoV RBD protein ...

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Abstract

The invention discloses a colloidal gold immunochromatographic test strip for detecting a neutralizing antibody secreted in vivo after injection of a novel coronavirus vaccine and a preparation methodof the colloidal gold immunochromatographic test strip. The test strip comprises a PVC bottom plate, and a sample pad, a colloidal gold pad, a nitrocellulose membrane and water absorption filter paper are sequentially lapped on the PVC bottom plate from left to right; the colloidal gold pad is coated with a recombinant novel coronavirus RBD protein marked by colloidal gold; and the nitrocellulosemembrane is coated with an angiotensin converting enzyme 2 (ACE2) detection line and coated with a mouse anti-chicken IgG antibody as a quality control line. According to the colloidal gold immunochromatography test strip, a competitive method is adopted to detect the neutralizing antibody in a human body, the sensitivity, specificity, repeatability and stability are high, the recovery rate of atarget compound is high, and the detection result is accurate and reliable. The test strip realizes rapid qualitative detection of the neutralizing antibody in a human body after injection of a novelcoronavirus vaccine, has high sensitivity and small intra-batch and inter-batch difference, and provides great convenience for clinical use.

Description

technical field [0001] The invention relates to a rapid qualitative detection test strip for blood samples and a preparation method thereof, in particular to a rapid qualitative detection test strip for detecting neutralizing antibodies in whole blood or serum after human injection of a new crown vaccine by using immune colloidal gold chromatography technology and its preparation method. Preparation. Background technique [0002] At present, there is no specific drug for COVID-19, and the new crown vaccine has become an effective means of preventing COVID-19. At present, my country's new crown vaccine research and development is generally in a leading position. Each technical route has vaccines that have entered the clinical stage. Among them, inactivated vaccines and adenovirus vector vaccines, a total of 4 vaccines from the two technical routes have entered the third phase of clinical trials. With the further success of follow-up vaccines, verifying the effectiveness of t...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569G01N33/558G01N33/58
CPCG01N33/6854G01N33/56983G01N33/558G01N33/587G01N2333/165G01N2469/20
Inventor 陈真诚申琳张启韩赵飞骏肖皓霖操良丽
Owner GUILIN UNIV OF ELECTRONIC TECH
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