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Immune chromatography test paper strip based on up-conversion luminescence technology

A technology of immunochromatographic test strips and test strips, which can be used in analytical materials, biological tests, material inspection products, etc., and can solve the problems of extremely harsh reaction conditions, large subjective influence, and low sensitivity.

Inactive Publication Date: 2010-07-28
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the fragility of the physical adsorption method (that is, the preparation of markers by hydrophobicity and electrostatic adsorption) makes the immunochromatography based on this marker extremely demanding for the reaction conditions, so some effective non-specific removal reagents, such as: Tween 20 (Tween 20), blow through 100 (Triton 100), sodium dodecyl sulfonate (SDS), etc., can only be used at low concentrations due to changes in hydrophobicity and electrostatic properties. The false positive rate and false negative rate of the immunochromatographic test paper of this kind of marker are inevitable results; in addition, the result of color judgment is easy to use and easy to operate, and it must be subject to the subjective influence of the observer and low sensitivity, and can only stay in the qualitative field. level, and absolutely impossible to achieve precise quantification
These shortcomings greatly limit the scope of application of immunochromatography in biological detection

Method used

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  • Immune chromatography test paper strip based on up-conversion luminescence technology

Examples

Experimental program
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Effect test

Embodiment 1

[0266] Example 1: Structure of upconversion luminescence biosensor

[0267] refer to Figure 26 and Figure 27 . As can be seen from the figure, the up-conversion luminescent biosensor of the present invention includes an excitation optical path, a phosphorescence image receiving optical path, and an image processing system, which are respectively used for illuminating the test strip 3, receiving the phosphorescence image emitted by the test strip 3, and receiving the phosphorescence image emitted by the test strip 3. Phosphorescence image analysis and processing, up-conversion luminescence biosensor pilot test strip structure diagram refer to Figure 24 .

[0268] On the excitation optical path, an infrared excitation light source 1 and a one-dimensional focusing mirror 2 are sequentially arranged along the optical axis, and the optical axis is 010. On the phosphorescent image receiving optical path, a front mirror group 5, a filter 6, a rear mirror group 7 and an image...

Embodiment 2

[0279] Example 2: Double-antibody sandwich detection of hepatitis B surface antigen HBs-Ag:

[0280] (1) Immunochromatography liquid phase material preparation:

[0281] A. UCP-antibody conjugates:

[0282] a. Use the established surface modification and activation methods to modify the surface of UCP particles with a diameter of 200-300nm, and link them with the anti-hepatitis B surface antigen HBs-Ag monoclonal antibody, and store them in UCP preservation solution (pH=7.20.03mol / L PB buffer, containing 0.1% BSA, 0.05% Tween20, 0.02% NaN 3 ) at a concentration of 1 mg / mL and stored at 4°C for later use;

[0283] b. Centrifuge 6 mL of 1 mg / mL UCP-antibody conjugate stored in UCP preservation solution at 12,000 r / min, 4°C for 30 min, and discard the supernatant as much as possible;

[0284] c. Settle the UCP-antibody conjugate in the centrifuge tube, add 3mL conjugate diluent (pH=7.20.03mol / LPB buffer, containing 1% sucrose, 1% BSA) and vortex to mix well (final concentrat...

Embodiment 3

[0327] Example 3: Double antibody sandwich detection of coronavirus (SARS virus):

[0328] (1) Immunochromatography liquid phase material preparation:

[0329] A. UCP-antibody conjugates:

[0330] a. Utilize established surface modification and activation method to carry out surface modification to the UCP particle of diameter 200-300nm, and link with the rabbit anti-SARS virus antibody of purification, in UCP storage solution (pH=7.20.03mol / L PB buffer solution Medium, containing 0.1% BSA, 0.05% Tween20, 0.02% NaN 3 ) at a concentration of 1 mg / mL and stored at 4°C for later use;

[0331] b. Centrifuge 6 mL of 1 mg / mL UCP-antibody conjugate stored in UCP preservation solution at 12,000 r / min, 4°C for 30 min, and discard the supernatant as much as possible;

[0332] c. Settle the UCP-antibody conjugate in the centrifuge tube, add 3mL conjugate diluent (pH=7.20.03mol / LPB buffer, containing 1% sucrose, 1% BSA) and vortex to mix well (final concentration: 2mg / mL UCP-antibod...

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Abstract

The invention discloses an immune chromatography test paper strip based on an up-conversion luminescence technology. The up-conversion luminescence material, i.e. UCP, is used as a biological marker, and the result can be presented in a visible light signal form under the radiation of infrared light and can be interpreted by an instrument, thereby quantitative detection on the target detected object is realized. The test paper strip mainly comprises a sample pad, a binding pad or a binder release pad, an analyzing membrane, a water absorbing pad and a plastic back plate. The invention also discloses a method for preparing the test paper strip, and application in the biological sample quantitative detection. According to different immune reaction modes of the substances to be detected, the test paper strip is classified according to a sandwich mode, a competition mode and an indirection mode, and various modes of the test paper strips can carry out quick and sensitive qualitative and quantitative detection and analysis on different substances to be detected in the sample.

Description

[0001] The patent application of the present invention is a divisional application, the original application number is 200410034104.0, the application date is April 23, 2004, and the invention name is immunochromatographic test strip based on up-conversion luminescence technology. technical field [0002] The invention relates to an immunochromatographic test strip, in particular to an immunochromatographic test strip using an up-conversion luminescent material, UCP, as a biomarker. The results of this kind of test strip are displayed in the form of visible light signals under the irradiation of infrared light, and can be interpreted by using up-changing luminescent biosensors, so as to realize rapid and sensitive qualitative and quantitative detection of target objects. Background technique [0003] Up-converting luminescent material (Up-Converting Phosphor, UCP) is a kind of rare earth metal compound that can convert energy up, that is, UCP can absorb low-energy (long-wavel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/532G01N33/547G01N33/52
Inventor 周蕾杨瑞馥王津郭兆彪郑岩侯秀洁
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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