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Immunochromatographic test paper for detecting human AIDS virus antibody and preparation method thereof

An immunochromatographic test strip, HIV technology, applied in coatings, measuring devices, analytical materials, etc., can solve the problems of inability to provide sufficient magnetic resonance signals, easy polymerization of magnetic beads, and long color development time.

Active Publication Date: 2016-05-11
昆明云大生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these preparation and modification methods are often cumbersome and complicated, and the obtained superparamagnetic composite particles cannot meet the requirements of LFIAs in terms of size, biocompatibility, saturation magnetic strength, external magnetic field response speed, stability, and labeling efficiency. The size of the beads is mostly larger than 300nm. Due to the large size of the magnetic beads, the swimming time on the test paper is slow and the color development time is long; and the particles that are too small cannot provide sufficient magnetic resonance signals; in addition, there are biocompatibility. Stability, easy aggregation of magnetic beads and other issues; these shortcomings limit the application of superparamagnetic composite particles in LFIAs

Method used

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  • Immunochromatographic test paper for detecting human AIDS virus antibody and preparation method thereof
  • Immunochromatographic test paper for detecting human AIDS virus antibody and preparation method thereof
  • Immunochromatographic test paper for detecting human AIDS virus antibody and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] (1) Preparation of recombinant protein A labeled with superparamagnetic composite particles

[0053] Superparamagnetic Fe with a particle size of 100nm, a particle size deviation of 15%, a magnetic saturation intensity of 40emu / g, a corresponding external magnetic field response speed of 20 seconds, and a surface carboxyl content of 80μmol / g was used. 3 o 4 Nanoparticles.

[0054] The specific method is: take 2.5mg of the above-mentioned superparamagnetic Fe 3 o 4 Nanoparticles were washed with MES buffer solution with a concentration of 0.1 mol and a pH of 4.7, and separated and enriched with a 0.4T magnetic rack, then resuspended with 1 ml of the above-mentioned MES buffer solution, and then added 0.96 mg of EDC and 2.17 mg of NHS, Mix well, react at a reaction temperature of 37° C. for 0.5 hour, and then wash with borax buffer solution with a concentration of 50 mmol and a pH of 8.5 to obtain 2.5 mg of activated magnetic particles.

[0055] Take 1.5 mg of recombi...

Embodiment 2

[0061] (1) Preparation of recombinant protein A labeled with superparamagnetic composite particles

[0062] Superparamagnetic Fe with a particle size of 60nm, a particle size deviation of 10%, a magnetic saturation intensity of 80emu / g, a corresponding external magnetic field response speed of 100 seconds, and a surface carboxyl content of 50μmol / g was used. 3 o 4 Nanoparticles.

[0063] The specific method is: take 2.5mg of the above-mentioned superparamagnetic Fe 3 o 4 Nanoparticles were washed with MES buffer solution with a concentration of 0.1 mol and a pH of 4.7, and separated and enriched with a 0.4T magnetic rack, then resuspended with 1 ml of the above-mentioned MES buffer solution, and then added 0.96 mg of EDC and 2.17 mg of NHS, Mix well, react at a reaction temperature of 35° C. for 40 minutes, and then wash with a borax buffer solution with a concentration of 50 mmol and a pH of 8.5 to obtain 2.5 mg of activated magnetic particles.

[0064] Take 1.5 mg of rec...

Embodiment 3

[0069] (1) Preparation of recombinant protein A labeled with superparamagnetic composite particles

[0070] Superparamagnetic Fe with a particle size of 300nm, a particle size deviation of 20%, a magnetic saturation intensity of 30emu / g, a corresponding external magnetic field response speed of 100 seconds, and a surface carboxyl content of 300μmol / g was used. 3 o 4 Nanoparticles.

[0071] The specific method is: take 2.5mg of the above-mentioned superparamagnetic Fe 3 o 4 Nanoparticles were washed with MES buffer solution with a concentration of 0.1 mol and a pH of 4.7, and separated and enriched with a 0.4T magnetic rack, then resuspended with 1 ml of the above-mentioned MES buffer solution, and then added 0.96 mg of EDC and 2.17 mg of NHS, Mix well, react at a reaction temperature of 37° C. for 20 minutes, and then wash with a borax buffer solution with a concentration of 50 mmol and a pH of 8.5 to obtain 2.5 mg of activated magnetic particles.

[0072] Take 1.5 mg of r...

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Abstract

The invention relates to an immunochromatographic test paper for detecting human immunodeficiency virus antibodies and a preparation method thereof, belonging to the field of reagents for detecting human immunodeficiency virus antibodies. The immunochromatographic test paper comprises a sample pad, a nitrocellulose membrane and a water absorption pad which are sequentially connected, wherein the sample pad contains a superparamagnetic composite particle labeled recombinant protein A; the nitrocellulose membrane is coated with a detection line and a quality control line which are mutually separate; the detection line contains a human immunodeficiency virus recombinant antigen; the quality control line contains an anti-recombinant protein A antibody which can be specifically combined with the recombinant protein A; and the human immunodeficiency virus is human immunodeficiency virus Type 1+2. The immunochromatographic test paper has the advantages of high sensitivity and high specificity, is quick and convenient, and can implement objective determination.

Description

technical field [0001] The invention belongs to the technical field of detecting human AIDS virus antibodies, and in particular relates to an immunochromatographic test paper for detecting human AIDS virus antibodies and a preparation method thereof. Background technique [0002] AIDS, Acquired Immunodeficiency Syndrome (AIDS), its pathogen is Human Immunodeficiency Virus (Human Immunodeficiency Virus, HIV), also known as AIDS virus, the virus destroys the body's immunity, causing the immune system to lose resistance, resulting in Various diseases and cancers can survive in the human body, and finally suffer from AIDS. At present, AIDS has not only become a public health problem that seriously threatens the health of our people, but also affects economic development and social stability. The detection of human immunodeficiency virus (HIV) 1+2 antibody is of great significance for the diagnosis and identification of AIDS, epidemiological investigation, screening of blood don...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/571G01N33/558G01N33/532
CPCG01N33/54333G01N33/558G01N33/56988G01N2446/86
Inventor 马岚
Owner 昆明云大生物技术有限公司
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