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Microfluidic paper based sensor based on fluoroimmunoassay and utilizing quantum dot as marker to detect norfloxacin in milk

A fluorescence immunoassay and norfloxacin technology, applied in the field of biological nanometers, can solve the problems of complex sample processing, difficult to popularize and high cost, and achieve the effects of wide application range, high sensitivity and low cost

Active Publication Date: 2017-12-22
NANJING UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem solved by the present invention is: aiming at the deficiencies in the prior art, a microfluidic paper-based sensor based on fluorescent immunoassay using quantum dots as a marker to detect norfloxacin in milk is proposed, which solves the problem of complicated sample processing and Disadvantages such as high cost and difficult promotion

Method used

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  • Microfluidic paper based sensor based on fluoroimmunoassay and utilizing quantum dot as marker to detect norfloxacin in milk
  • Microfluidic paper based sensor based on fluoroimmunoassay and utilizing quantum dot as marker to detect norfloxacin in milk
  • Microfluidic paper based sensor based on fluoroimmunoassay and utilizing quantum dot as marker to detect norfloxacin in milk

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Embodiment 1

[0031] A microfluidic paper-based sensor for detecting norfloxacin in milk based on fluorescent immunoassay using quantum dots as a marker, comprising the following steps:

[0032] a. Formation of plasma treated paper

[0033] 1) Design a paper-based model of a certain size, and form a hydrophilic area (such as figure 1 Shown in white area) and hydrophobic area (such as figure 1 shown in black area);

[0034] 2) Bake at 105°C for 5 minutes;

[0035] 3) Treat the paper with a plasma cleaner for 4 minutes to obtain the required paper;

[0036] 4) The plasma-treated paper needs to be sealed before adding samples to prevent oxidation.

[0037] b. Preparation of NOR-BSA coated antigen

[0038] 1) Add 11.7mgNOR to 3mL of anhydrous dimethylformamide;

[0039] 2) Add 70.5mgEDC.HCl and 21.2mgNHS;

[0040] 3) Centrifuge after stirring overnight at room temperature;

[0041] 4) The supernatant was added to 33.0 mg of BSA dissolved in 5 mL of PBS (0.01 M, pH=7.4);

[0042] 5) Aft...

Embodiment 2

[0060] A microfluidic paper-based sensor for detecting norfloxacin in milk based on fluorescent immunoassay using quantum dots as a marker, comprising the following steps:

[0061] a. Formation of plasma treated paper

[0062] 1) Design a paper-based model of a certain size, and form a hydrophilic area (such as figure 1 Shown in white area) and hydrophobic area (such as figure 1 shown in black area);

[0063] 2) Bake at 105°C for 5 minutes;

[0064] 3) Treat the paper with a plasma cleaner for 4 minutes to obtain the required paper;

[0065] 4) The plasma-treated paper needs to be sealed before adding samples to prevent oxidation.

[0066] b. Preparation of NOR-BSA coated antigen

[0067] 1) Add 11.7mgNOR to 3mL of anhydrous dimethylformamide;

[0068] 6) Add 70.5mgEDC.HCl and 21.2mgNHS;

[0069] 7) Centrifuge after stirring overnight at room temperature;

[0070] 8) The supernatant was added to 33.0 mg of BSA dissolved in 5 mL of PBS (0.01 M, pH=7.4);

[0071] 9) Aft...

Embodiment 3

[0083] A microfluidic paper-based sensor for detecting norfloxacin in milk based on fluorescent immunoassay using quantum dots as a marker, comprising the following steps:

[0084] a. Formation of plasma treated paper

[0085] 1) Design a paper-based model of a certain size, and form a hydrophilic area (such as figure 1 Shown in white area) and hydrophobic area (such as figure 1 shown in black area);

[0086] 2) Bake at 105°C for 5 minutes;

[0087] 3) Treat the paper with a plasma cleaner for 4 minutes to obtain the required paper;

[0088] 4) The plasma-treated paper needs to be sealed before adding samples to prevent oxidation.

[0089] b. Preparation of NOR-BSA coated antigen

[0090] 1) Add 11.7mgNOR to 3mL of anhydrous dimethylformamide;

[0091] 10) Add 70.5mgEDC.HCl and 21.2mgNHS;

[0092] 11) Centrifuge after stirring overnight at room temperature;

[0093] 12) The supernatant was added to 33.0 mg of BSA dissolved in 5 mL of PBS (0.01 M, pH=7.4);

[0094] 13)...

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Abstract

The invention discloses a microfluidic paper based sensor based on fluoroimmunoassay and utilizing a quantum dot as a marker to detect norfloxacin (NOR) in milk and belongs to the technical field of biological nanometer. A preparation method of the sensor comprises the steps: firstly, forming a hydrophilic area and a hydrophobic area on chromatographic paper through a wax spray printing technology, roasting for a certain time and performing plasma treatment on the paper; then, preparing NOR-bovine serum albumin (BSA) coating antigen and a quantum dot marked NOR antibody; then, respectively adding the coating antigen and a goat anti-mouse secondary antibody into two white paper areas in parallel up and down and adding an NOR and quantum dot marked antibody mixture into a coating layer, folding the paper and permeating the solution into a secondary antibody area; finally, utilizing immunoassay determination to finally quantitatively detect the norfloxacin in the milk through fluorescent signals. The microfluidic paper based sensor disclosed by the invention has the advantages of low cost, simpleness in operation, high flexibility, low detection limit and quickness in detecting NOR.

Description

technical field [0001] The invention relates to biological materials and nanomaterials, and belongs to the field of biological nanotechnology, in particular to a microfluidic paper-based sensor for detecting norfloxacin in milk based on fluorescent immunoassays using quantum dots as markers. Background technique [0002] Norfloxacin is an antimicrobial drug widely used in human and veterinary therapy, which has the characteristics of broad antibacterial spectrum and strong antibacterial activity. However, residues in waste water, surface water, food, etc. or long-term abuse of norfloxacin will cause serious harm to humans and animals. Therefore the detection of norfloxacin has become an increasingly important issue. At present, many methods for detecting norfloxacin have been reported in the literature, mainly including high performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), fluorescence method, ultraviolet photometry, etc. However, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/533
CPCG01N21/6428G01N33/533
Inventor 于海东宗丽君李林张承武刘志鹏刘金华黄维欧阳启然赖琼宇朱成显焦钰翠高磊
Owner NANJING UNIV OF TECH
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