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Immunofiltration assay fluorescent quantitative detection method based on high-sensitivity quantum dot

A fluorescence quantitative detection and immunodiafiltration technology, applied in the field of fluorescence detection, can solve the problems of insufficient quantitative sensitivity and linear range of colloidal gold, and achieve the effects of low cost, simple method and good luminescence stability.

Active Publication Date: 2014-06-25
BEIJING KANGMEI TIANHONG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Compared with fluorescence detection, colloidal gold quantification is insufficient in sensitivity and linear range

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1: Double-antibody sandwich determination of human chorionic gonadotropin (hCG)

[0051] (1) Synthesis of water-soluble quantum dots by aqueous phase method

[0052] For the synthesis of CdTe quantum dots, add absolute ethanol and primary water to the reactor containing appropriate amount of Te powder and sodium borohydride in the ratio of 2:1 by volume, and react in a water bath at 60°C under nitrogen gas until black Te The powder is completely gone. Add a slight excess of 0.5mol / L H 2 SO 4 , yielding H 2 Te is absorbed by NaOH solution to obtain NaHTe aqueous solution.

[0053] CdCl under nitrogen 2 2.5H 2 O was dissolved in 100mL ultrapure water, then added mercaptopropionic acid, and 0.5mol / L NaOH solution was added dropwise under stirring to adjust the pH of the solution to 11.2, and finally 0.1mmol NaHTe solution was injected, wherein Cd: Te: mercaptopropionic acid The ratio is 1.0:0.5:2.4. The resulting mixed solution was heated to reflux at 100° ...

Embodiment 2

[0068] Embodiment 2: Double-antigen sandwich determination of Helicobacter pylori antibody (HP)

[0069] (1) Synthesis of fat-soluble quantum dots and water-soluble modification by organic phase method

[0070] Take 0.2375g of selenium powder, 2.60mL of octadecene and 1.57mL of tri-n-octylphosphine, add them in turn to a 25ml small reagent bottle, heat the mixed liquid in the bottle and shake repeatedly until the selenium powder is completely dissolved, and then the selenium precursor is obtained. Another 0.0368g of cadmium oxide, 0.342g of stearic acid and 3.8ml of octadecene were added to the three-necked flask in sequence, and heated under the protection of nitrogen until the cadmium oxide was completely dissolved. Lower the temperature to cool the solution to solidify. Take 2.25g of octadecylamine and 0.95g of trioctylphosphine oxide, put them into a three-necked flask, and heat to melt the solid, continue to heat to 280°C, inject 4.2mL of selenium precursor, and raise th...

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Abstract

The invention discloses an immunofiltration assay fluorescent quantitative detection method based on a high-sensitivity quantum dot; the immunofiltration assay fluorescent quantitative detection method comprises the following steps of: constructing a fluorescence immunofiltration array device by using the excellent fluorescent characteristic of the quantum dot in combination of a quantum dot fluorescence labeling technology and an immunofiltration array technology on the basis of optimizing constituent parts for the immunofiltration assay; and after immunofiltration array, detecting the strenght of fluorescent signals of the quantum dot and a quality control dot by using a fluorescence quantometer, correcting the fluorescence strenght of the quantum pot by using the quality control dot, and further realizing the quantitative detection of a tested object according to a standard curve obtained by using the fluorescence quantometer. The method is simple, rapid, accurate, low in cost and high in sensitiveness. Comapred with the conventional collodial gold immunofiltration array method, the immunofiltration assay fluorescent quantitative detection method has the advantages of good labeling stability, low non-specificity, high sensitivity, wide linear range and accurate quantification. The method is suitable for samples such as serums, urine, spittle, excrement and the like and can be applicable to the detection of serious illness, poisons, food safety and the like.

Description

technical field [0001] The invention discloses a quantum dot-based immunofiltration fluorescence quantitative detection method, which can realize the quantitative detection of target analytes such as pathogens, major diseases (such as tumors, cardiovascular diseases, etc.), drug detection, drug and food safety, etc. The invention belongs to the technical field of fluorescence detection. Background technique [0002] Immunofiltration (IFA) is a common method in clinical testing. Its basic principle is: using a nitrocellulose (NC) membrane as a carrier, using the filterability of the microporous membrane, the antigen-antibody reaction and Washing is done quickly by liquid percolating through the membrane on a special diafiltration device. The immunofiltration assay was originally developed and established on the basis of the dot ELISA, and the conjugates used were enzyme-labeled. The diafiltration device is one of the main reagent components in IFA, which consists of three p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/558G01N21/64
Inventor 王东
Owner BEIJING KANGMEI TIANHONG BIOTECH
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