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Colloidal gold immunochromatography test strip for simultaneously detecting viral antigens and antibodies

An immunochromatographic test strip and virus antigen technology, which is applied in measurement devices, analytical materials, instruments, etc., can solve the problem of not detecting influenza virus antigens at the same time, and achieve fast and intuitive reading results, low price and strong specificity. Effect

Inactive Publication Date: 2014-08-13
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Unfortunately, there is currently no test strip that simultaneously detects influenza virus antigens and their specific IgM and IgG antibodies

Method used

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  • Colloidal gold immunochromatography test strip for simultaneously detecting viral antigens and antibodies
  • Colloidal gold immunochromatography test strip for simultaneously detecting viral antigens and antibodies
  • Colloidal gold immunochromatography test strip for simultaneously detecting viral antigens and antibodies

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1: Influenza A virus detection

[0048] Detection of Influenza A Virus Antigen:

[0049] (1) Take 200 μg of influenza A labeled antibody and place it in a dialysis bag for dialysis against 5mM Tris-HCl for 24 hours. During this process, change the water every 2 hours. Ultrapure water to 2mL, discard precipitated impurities after centrifugation.

[0050] (2) Preparation of gold-labeled antibody conjugate pads: colloidal gold was prepared by reducing chloroauric acid with sodium citrate, and the glass instruments used were pre-soaked with washing solution overnight, and then rinsed with deionized water. Add 1000mL of ultrapure water to the beaker, then add 10mL of 1% sodium citrate solution, heat to boiling, then add 10mL of 1% chloroauric acid solution, boil for 15min, cool down, and store at 4°C. The particle diameter is about 20-30nm (such as Figure 14 ), put 20mL colloidal gold solution in a beaker, add 200μL 0.01M K 2 CO 3 Solution Adjust the pH of t...

Embodiment 2

[0059] Embodiment 2: Influenza B virus detection

[0060] Detection of influenza B virus antigen:

[0061] (1) Take 200 μg of influenza B labeled antibody in a dialysis bag and dialyze against 5mM Tris-HCl for 24 hours. During this process, change the water every 2 hours. After the dialysis is completed, take out the antibody and place it in a centrifuge tube, add Ultrapure water to 2mL, discard precipitated impurities after centrifugation.

[0062] (2) Preparation of gold-labeled antibody conjugate pads: colloidal gold was prepared by reducing chloroauric acid with sodium citrate, and the glass instruments used were pre-soaked with washing solution overnight, and then rinsed with deionized water. Add 1000mL of ultrapure water to the beaker, then add 10mL of 1% sodium citrate solution, heat to boiling, then add 10mL of 1% chloroauric acid solution, boil for 15min, cool down, and store at 4°C. The particle diameter is about 20-30nm, take 20mL colloidal gold solution and put i...

Embodiment 3

[0070] Embodiment 3: detection of adenovirus

[0071] Detection of Adenovirus Antigen:

[0072] (1) Take 200 μg of labeled antibody against adenovirus and place it in a dialysis bag for dialysis against 5mM Tris-HCl for 24 hours. During this process, change the water every 2 hours. After the dialysis is completed, take out the antibody and place it in a centrifuge tube. Ultrapure water to 2mL, discard precipitated impurities after centrifugation.

[0073] (2) Preparation of gold-labeled antibody conjugate pads: colloidal gold was prepared by reducing chloroauric acid with sodium citrate, and the glass instruments used were pre-soaked with washing solution overnight, and then rinsed with deionized water. Add 1000mL of ultrapure water to the beaker, then add 10mL of 1% sodium citrate solution, heat to boiling, then add 10mL of 1% chloroauric acid solution, boil for 15min, cool down, and store at 4°C. The particle diameter is about 20-30nm, take 20mL colloidal gold solution and...

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Abstract

The invention relates to a colloidal gold immunochromatography test strip for simultaneously detecting viral antigens and antibodies. The test strip comprises a first glue strip and a second glue strip which are arranged in parallel, wherein the first glue strip comprises a first sample pad, a first conjugate pad, a first nitrocellulose membrane and first absorbent paper, which are sequentially lapped with one another, the first conjugate pad is coated with colloidal gold marked by the viral antigens, the first nitrocellulose membrane is coated with viral antibodies and antibodies, the viral antibodies are taken as a first detection line, and the antibodies are used for resisting the viral antigens on the colloidal gold and are taken as a quality control line; the second glue strip comprises a second sample pad, a second conjugate pad, a second nitrocellulose membrane and second absorbent paper, which are sequentially lapped with one another, the second conjugate pad is coated with the colloidal gold marked by the viral antigens, the second nitrocellulose membrane is coated with anti-human IgG antibodies and anti-human IgM antibodies, the anti-human IgG antibodies are taken as a second detection line, and the anti-human IgM antibodies are taken as a third detection line. The colloidal gold immunochromatography test strip can be used for simultaneously detecting the viral antigens as well as the IgG antibodies and IgM antibodies of the viral antigens.

Description

technical field [0001] The invention belongs to the technical field of immune detection and analysis, and relates to a colloidal gold immunochromatographic test strip for simultaneously detecting virus antigens and their specific immunoglobulin M (IgM) and immunoglobulin G (IgG). Background technique [0002] Influenza (abbreviated as influenza) is an acute respiratory infection caused by influenza virus, and it is also a disease with strong contagion and fast transmission speed. It is mainly spread through droplets in the air, person-to-person contact or contact with contaminated items. Typical clinical symptoms are: sudden onset of high fever, general pain, significant fatigue and mild respiratory symptoms. Generally, autumn and winter are the high-incidence periods, and the complications and deaths caused by them are very serious. The disease is caused by influenza virus, which belongs to the Orthomyxoviridae family, with a diameter of 80-120nm, spherical or filamentous...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/577
CPCG01N33/56983G01N33/558G01N33/6893
Inventor 蒋兴宇曹丰晶张伟陈翊平赵大龙
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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