Method for improving sensitivity of immunological detection system and device using method

An immune detection and sensitivity technology, applied in the field of a method and a device using the method, can solve the problems of no practical application value, inability to distinguish between negative and positive, and achieve the effects of high sensitivity, improved sensitivity, and enhanced detection signal

Pending Publication Date: 2018-01-30
广州佰芮慷生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But, in practical application, it is impossible to avoid the generation of non-specific reaction (such as: background signal), therefore, these methods can't distinguish negative and positive in practical application, and final result is that all samples are all positive, therefore, these methods no practical value

Method used

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  • Method for improving sensitivity of immunological detection system and device using method
  • Method for improving sensitivity of immunological detection system and device using method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 A kind of method that improves the immunochromatography test strip sensitivity of detecting HCV core protein, comprises:

[0025] 1. Preparation of nano-gold labeled detection antibody: take 10 mL of nano-gold particles with a particle size of 40 nm, add 10 μg of HCVcore protein detection antibody; react at room temperature for 15 min; add BSA to a final concentration of 1% (W / V), continue to react at room temperature for 15 min; centrifuge , resuspend the particles with 1 mL of reconstitution solution (10 mM Tris-HCl (pH 8.0), 0.5% BSA), and set aside.

[0026] 2. Preparation of biotin-labeled capture antibody: prepare biotin-labeled HCV core protein capture antibody according to the coupling ratio of capture antibody and Sulfo-NHS-LC-LC-Biotin of 1:1 to 1:40; use PBS (pH7. 2) Dialyze 3 times to remove impurities and set aside.

[0027] 3. Preparation of chromatographic membrane: Spray the anti-biotin monoclonal antibody (detection line) with a concentrat...

Embodiment 2

[0034] Embodiment 2 A method for improving the sensitivity of immunochromatographic test strips for detecting HIV p24 protein, comprising:

[0035] 1. Preparation of nano-gold labeled detection antibody: Take 10 mL of nano-gold particles with a particle size of 40 nm, add 10 μg of HIVP24 protein detection antibody; react at room temperature for 15 min; add BSA to a final concentration of 1% (W / V), continue to react at room temperature for 15 min; centrifuge , resuspend the particles with 1 mL of reconstitution solution (10 mM Tris-HCl (pH 8.0), 0.5% BSA), and set aside.

[0036] 2. Preparation of biotin-labeled capture antibody: prepare biotin-labeled HIV P24 protein capture antibody according to the coupling ratio of capture antibody to Sulfo-NHS-LC-LC-Biotin of 1:16; dialyze with PBS (pH7.2) for 3 The second time, remove impurities, and dilute the biotin-labeled HIV P24 protein capture antibody to 0.25 mg / mL with PBS for later use.

[0037]3. Preparation of chromatographic ...

Embodiment 3

[0044] Embodiment 3 A method for improving the sensitivity of the immunofiltration card for detecting HIV p24 protein, comprising:

[0045] 1. Preparation of nano-gold labeled detection antibody: take 10 mL of nano-gold particles with a particle size of 40 nm, add 10 μg of HIVp24 protein detection antibody; react at room temperature for 15 minutes; add BSA to a final concentration of 1% (W / V), and continue to react at room temperature for 15 minutes; Centrifuge, resuspend the particles with 1 mL of reconstitution solution (10 mM Tris-HCl (pH 8.0), 0.5% BSA), and set aside.

[0046] 2. Preparation of FITC (fluorescein isothiocyanate)-labeled capture antibody: prepare FITC-labeled HIV p24 protein capture antibody according to the coupling ratio of capture antibody to NHS-FITC of 1:15; dialyze with PBS (pH7.2) for 3 Second, remove impurities and set aside.

[0047] 3. Preparation of the diafiltration membrane: On the nitrocellulose membrane, the anti-FITC monoclonal antibody wit...

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Abstract

The invention discloses a novel immunological detection method. A dendritic 'signal labeled detection antibody-target antigen-small molecule labeled capture antibody-target antigen-signal labeled detection antibody-target antigen-small molecule labeled capture antibody...' composition is formed and fixed to a detection area by a small molecule antibody on a solid phase, so that good signal amplification effects are achieved, and sensitivity is greatly improved. Compared with an existing signal amplification technology, the novel immunological detection method has the advantages that detectionsignals can be enhanced only under the condition of presence of target antigen, and accordingly, high detection sensitivity is ensured while specificity is ensured.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a method for improving the sensitivity of an immune detection system and a device using the method. Background technique [0002] In 1959, Yalow and Berson established radioimmunoassay by combining radioactive isotope tracer and immune reaction, creating a new field of immunoassay, which was recognized as a major breakthrough in trace analytical chemistry. Since then, a series of immunoassay methods have emerged and continued to develop, such as enzyme-linked immunosorbent assay, fluorescent immunoassay, immunochromatographic assay, and immunosensor assay. Immunoassay technology is developing in the direction of quantitative, multi-component analysis, high sensitivity, high specificity, convenience, rapidity and more economical application. [0003] Simple, sensitive, and effective detection technology is of great significance in the fields of disease diagnosis, new drug deve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/577
Inventor 张玲其他发明人请求不公开姓名
Owner 广州佰芮慷生物科技有限公司
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