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Paper strip for quantitative electrochemical biological test

A quantitative detection and test strip technology, which is applied in biological testing, material electrochemical variables, microbial measurement/inspection, etc., can solve the problems of limited immobilization of biomolecules and affecting detection sensitivity, etc.

Inactive Publication Date: 2007-02-07
郭良宏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the description of US Patent 6670115, the test strip is only used as a reagent carrier, and the immune reaction and signal detection are carried out on the electrode surface. This method has a limited immobilization amount of biomolecules, which affects the detection sensitivity.

Method used

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  • Paper strip for quantitative electrochemical biological test
  • Paper strip for quantitative electrochemical biological test
  • Paper strip for quantitative electrochemical biological test

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1: Electrochemical method to detect the immune reaction between mouse IgG / goat anti-mouse IgG on nitrocellulose membrane.

[0017] Take a square nitrocellulose membrane with a side length of 1 cm and soak it overnight in 10 μg / mL mouse IgG (dissolved in 20 mM sodium carbonate / sodium bicarbonate buffer, pH 9.5). After washing, block with 1% BSA for 2 hours. Add different concentrations of alkaline phosphatase-labeled goat anti-mouse antibody, and shake at room temperature for 1 hour. After cleaning, the membrane was placed on the screen-printed electrode sheet and phosphate phenol was added. After waiting for 5 minutes, start the cyclic voltammetry detection program of the electrochemical workstation, and record the current signal. get as figure 2 The results shown show that the detected current signal has a linear relationship with the concentration of the enzyme-labeled antibody within a certain range, and the lowest level of 0.1 μg / mL (0.7nM) antibody can ...

Embodiment 2

[0018] Example 2: Electrochemical method for detection of immune reaction between estradiol antigen / antibody on nitrocellulose membrane.

[0019] Take a square nitrocellulose membrane with a side length of 1 cm and soak it overnight in 10 μg / mL estradiol antigen (dissolved in 20 mM sodium carbonate / sodium bicarbonate buffer, pH 9.5). After washing, block with 1% BSA for 1 hour. Different concentrations of estradiol mouse monoclonal antibody were added, and the reaction was shaken at room temperature for 1 hour. After washing, add alkaline phosphatase-labeled goat anti-mouse secondary antibody diluted 1000 times and react with shaking at room temperature for 1 hour. After cleaning, the membrane was placed on the screen-printed electrode sheet and phosphate phenol was added. After waiting for 5 minutes, start the cyclic voltammetry detection program of the electrochemical workstation, and record the current signal. get as image 3 The results shown indicate that the detected...

Embodiment 3

[0020] Example 3: Electrochemical method to detect the competitive immune response of estradiol on nitrocellulose membrane.

[0021] Take a square nitrocellulose membrane with a side length of 1 cm and soak it overnight in 10 μg / mL estradiol antigen (dissolved in 20 mM sodium carbonate / sodium bicarbonate buffer, pH 9.5). After washing, block with 1% BSA for 1 hour. Add different concentrations of estradiol and 0.1 μg / mL estradiol mouse monoclonal antibody, and shake at room temperature for 1 hour. After washing, add alkaline phosphatase-labeled goat anti-mouse secondary antibody diluted 1000 times and react with shaking at room temperature for 1 hour. After cleaning, the membrane was placed on the screen-printed electrode sheet and phosphate phenol was added. After waiting for 5 minutes, start the cyclic voltammetry detection program of the electrochemical workstation, and record the current signal. get as Figure 4 The results shown indicate that the detected current sign...

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PUM

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Abstract

The disclosed quantitative detective test paper for bio-combination reaction is prepared as following: based on solid substrate, preparing electrode firstly; adding a layer of film with fixed catching molecules, wherein the target molecule competes with the catching molecule and another enzyme-mark molecule to combine together, and it uses the electrochemical signal of reaction product for detection. This invention is simple and fast, and has wide application.

Description

technical field [0001] The invention relates to a test paper strip for quantitatively detecting biological binding reactions through an electrochemical method. Background technique [0002] Qualitative and quantitative analysis of many substances can be carried out through antigen / antibody immunoassay. These substances include small organic molecules, amino acids, polypeptides, polysaccharides, and biological macromolecules such as proteins, nucleic acids, viruses, and bacteria. Generally speaking, immunoassays should be operated in special laboratories, using microplates or test tubes as reaction vessels, and after a long period of reaction, a lower detection limit can be reached. The main disadvantage of conventional immunoassays is that due to the multi-step operations involved, including antigen / antibody reaction, cleaning, enzyme / substrate reaction, etc., the analysis time is long, the process is complicated, and special instruments and equipment are required, so it is ...

Claims

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Application Information

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IPC IPC(8): G01N27/406G01N33/50G01N27/416G01N27/30C12Q1/00
Inventor 郭良宏
Owner 郭良宏
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