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Construction and application of chemiluminiscence-colloidal gold immunochromatography test paper

An immunochromatographic test paper and chemiluminescent technology, which is applied in scientific instruments, analytical materials, biological testing, etc., can solve the problems of low detection sensitivity, long time-consuming, complicated detection steps, etc., and achieve large specific surface area, high sensitivity, and optical properties. big effect

Active Publication Date: 2014-10-01
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The microbial detection method is simple and easy to implement, but due to its low detection sensitivity and long time-consuming, it cannot meet the requirements of on-site and rapid detection
CN102353779A discloses a microbial detection method, which still cannot meet the requirements of high sensitivity and rapid detection
This method still cannot realize qualitative and quantitative detection at the same time, and the detection steps are complicated

Method used

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  • Construction and application of chemiluminiscence-colloidal gold immunochromatography test paper
  • Construction and application of chemiluminiscence-colloidal gold immunochromatography test paper
  • Construction and application of chemiluminiscence-colloidal gold immunochromatography test paper

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Experimental program
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Effect test

Embodiment 1

[0058] figure 1 and figure 2 They are the schematic diagram of the immunochromatography test strip and the construction principle diagram of the chemiluminescence-colloidal gold immunochromatography test strip, respectively. The specific steps for the construction of the chemiluminescent-colloidal gold immunochromatographic test strip are as follows:

[0059] 1) Preparation of colloidal gold-enzyme-antibody complex:

[0060] Adjust the pH value of 10mL of colloidal gold solution to 8.5, then add 0.01mg of detection antibody and 0.2mg of horseradish peroxidase (HRP), the coupling ratio of the enzyme and antibody is 10:1, at room temperature Gently shake and react for 1-2 hours, then add 3mL of 3% BSA solution to block the excess sites, react for 0.5-1 hour, then centrifuge for 10-20 minutes at a centrifugal force of 10000-12000r / min, remove the upper Add 2 mL of PBS solution containing 0.1% BSA with a concentration of 0.01M and a pH value of 7.4, resuspend, and then centrif...

Embodiment 2

[0066] Detection of alpha-fetoprotein (AFP).

[0067] The experimental steps are as follows:

[0068] (1) Use PBS buffer solution to construct AFP solutions with concentrations of 200ng / mL, 100ng / mL, 50ng / mL, 20ng / mL, 10ng / mL, 5ng / mL, 1ng / mL and 0ng / mL, and then add them to Chemiluminescence-colloidal gold immunochromatography test strips, immunochromatography reaction 5-10min;

[0069] (2) Observe the color depth of the colloidal gold on the test strip with the naked eye, and take pictures with a camera to obtain the result of visual detection of AFP;

[0070] (3) Add a chemiluminescent substrate to the original test strip, and quantitatively detect the result with a chemiluminescent instrument.

[0071] image 3 It is the result of visual detection of alpha-fetoprotein (AFP) by chemiluminescence-colloidal gold test strips, from image 3 It can be seen from the color of the colloidal gold that the minimum detection limit of the test strip for AFP is 10ng / mL.

[0072] F...

Embodiment 3

[0075] Detection of fetal cancer protein (CEA) in serum.

[0076] The experimental steps are as follows:

[0077] (1) Use PBS buffer solution to construct CEA solutions with concentrations of 200ng / mL, 100ng / mL, 50ng / mL, 20ng / mL, 10ng / mL, 5ng / mL, 1ng / mL and 0ng / mL, and then add them to Chemiluminescence-colloidal gold test strips, immunochromatographic reaction for 5-10min;

[0078] (2) Observe the color depth of the colloidal gold on the test strip with the naked eye, and take pictures with a camera to obtain the result of visual detection of CEA;

[0079] (3) Add a chemiluminescent substrate to the original test strip, and quantitatively detect the result with a chemiluminescent instrument.

[0080] Figure 5 It is the result of visual detection of Fetal Oncoprotein (CEA) by chemiluminescence-colloidal gold test strips, from Figure 5 It can be seen from the color of the colloidal gold that the minimum detection limit of the test strip for CEA is 10 ng / mL.

[0081] Fi...

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Abstract

The invention relates to a construction method and an application of chemiluminiscence-colloidal gold immunochromatography test paper, which has high sensitivity and is quantitative. The method comprises the following steps: chemiluminiscence and a colloidal gold immunochromatography method are combined, namely the surface of colloidal gold is simultaneously coupled with a detection antibody and an enzyme to form a colloidal gold-enzyme-antibody compound; the colloidal gold-enzyme-antibody compound and a coated antibody are sequentially sprayed on a sample combination cushion and a film; an immunochromatography reaction is carried out and qualitative detection is realized based on color shades of the colloidal gold; quantitative detection is carried out based on the light intensity of an enzyme catalyzed chemiluminiscence reaction so as to construct the chemiluminiscence-colloidal gold immunochromatography test paper; the test paper is applied to the detection of biological macromolecules and micro-molecules. The chemiluminiscence-colloidal gold immunochromatography test paper can simultaneously realize quantitative, qualitative and high-sensitivity detection and is suitable for field detection and domestic individualized diagnosis.

Description

technical field [0001] The invention relates to the field of immunological detection and analysis, in particular to a construction method and application of an immunochromatographic test strip, in particular to the construction and application of a chemiluminescent-colloidal gold immunochromatographic test strip. Background technique [0002] Achieving rapid, sensitive and low-cost detection and diagnosis of pesticide and veterinary drug residues, infectious animal and plant pathogens, viruses, and targets in clinical samples is of great significance to the protection of human health and life safety, the protection of the environment and national security, and the maintenance of social stability. Significance. [0003] At present, relatively mature detection methods mainly include instrumental analysis, microbiological detection, molecular biology and immunological detection and other methods. [0004] The instrumental analysis method has the advantages of high sensitivity ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/68
CPCG01N33/521G01N33/558G01N33/6803
Inventor 蒋兴宇陈翊平曹丰晶张晓青张伟查瑞涛
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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