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Test strip for detecting tetanus antibody on basis of quantum dot fluorescent microsphere chromatography and preparation method and using method thereof

A technology of fluorescent microspheres and quantum dots, used in biological testing, measuring devices, analytical materials, etc., to achieve the effects of good stability, improved sensitivity and simple operation

Active Publication Date: 2019-01-15
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention proposes a test strip for detecting tetanus antibody based on quantum dot fluorescent microsphere chromatography and its preparation method and use method, which solves the technical problems encountered in fast, low-consumption and portable detection of tetanus antibody content

Method used

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  • Test strip for detecting tetanus antibody on basis of quantum dot fluorescent microsphere chromatography and preparation method and using method thereof
  • Test strip for detecting tetanus antibody on basis of quantum dot fluorescent microsphere chromatography and preparation method and using method thereof
  • Test strip for detecting tetanus antibody on basis of quantum dot fluorescent microsphere chromatography and preparation method and using method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0031] The preparation of quantum dot fluorescent microsphere-labeled protein, the steps are as follows:

[0032] (1) First, 100 μL of carboxylated quantum dot microspheres (QDNB) was centrifuged at 15000 for 30 min, and then redissolved in 100 μL of 50 mM pH 6.0 MES buffer. Add 15 μL of 8 mM N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC), and rotate in the dark room for 30 min to activate the quantum dot microspheres, and then Centrifuge at 15000 for 30 min.

[0033] (2) Use 100 μL of 50 mM pH 7.4 boric acid buffer for ultrasonic reconstitution, then add different volumes of 5 mg / ml Fc, respectively 2 μL, 3 μL, 5 μL, 7 μL, and 10 μL, rotate in the dark room for 60 min, and then Perform centrifugation at 15000 for 30min.

[0034](3) Add 100 μL of 2% BSA blocking solution to redissolve and perform blocking treatment. After 50 min of darkroom rotation reaction, centrifuge at 15000 for 30 min. The final prepared QDNB-Fc is stored in 100 μL of boric acid buffe...

Embodiment 2

[0037] The preparation method of the test strip for detecting tetanus antibody based on quantum dot fluorescent microsphere chromatography, the steps are as follows: the sample pad is first blocked with a mass concentration of 0.5-5% TW20 solution, and then the detection line is drawn on the NC membrane and For the quality control line, vacuum-dry the treated sample pad and NC film at 37°C for 1 hour, and then paste the sample pad, NC film and water-absorbent pad on the PVC bottom plate in sequence, overlapping each other by 2-3mm, and finally cut into 3.9mm Width, that is, the finished test strip.

[0038] Detection of tetanus antibody by fluorescent microsphere immunochromatographic test strip:

[0039] The test strips prepared by the above steps are prepared with a certain concentration gradient of tetanus antibody standard solution (0-0.022IU), and the concentration gradient of tetanus antibody standard solution is mixed with a certain amount of QDNB-Fc respectively, and t...

Embodiment 3

[0041] Interference of different types of antibodies on tetanus antibody detection:

[0042] Prepare the complex of quantum dot microsphere antibody and the test strip coated with antigen through the above steps, then configure the concentration of tetanus antibody standard solution to be 0.01 IU, and the concentration of interfering antibody to be 10 equivalents. For example, the concentration of pertussis antibody is 0.1 IU, 0.1 IU for rabies antibody, 5 μg / ml for cytokeratin antibody, 5 μg / ml for thyroid transcription factor antibody, mixed solution of pertussis antibody with a concentration of 0.1 IU and tetanus antibody with a concentration of 0.01 IU. After the sample was mixed with a certain amount of QDNB-Fc, 90 μL of the mixed solution was dropped on the sample pad of the tetanus antibody test strip. After 10 min, the fluorescence peak intensities corresponding to different antibodies were recorded, and each experiment was repeated 3 times in parallel. Such as Figur...

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Abstract

The invention belongs to the technical field of lateral side stream chromatography and bioanalysis, particularly relates to a test strip for detecting tetanus antibody on basis of quantum dot fluorescent microsphere chromatography and a preparation method and using method thereof, and aims at solving the technical problems in the process of rapidly and portably detecting tetanus antibody content via low consumption. The preparation method of the test strip comprises the following steps of: firstly closing a sample pad by using solution with a TW20 mass concentration of 0.5-5%; marking a detection line and a quality control line on an NC membrane in sequence; drying the processed sample pad and the NC membrane at 37 DEG C for 1 hour; pasting the sample pad, the NC membrane and a water absorption pad on a PVC baseboard in sequence; carrying out overlapping in pairs to 2-3 mm; and finally cutting into 3.9 mm wide to obtain the finished test strip. Compared with traditional colloidal goldimmunochromatography test strips, the test strip is good in quantum dot microsphere marker stability and high in detection sensitivity, and is capable of realizing quantitative detection for low-concentration target objects.

Description

technical field [0001] The invention belongs to the technical fields of lateral flow chromatography and bioanalysis, and in particular refers to a test strip for detecting tetanus antibody based on quantum dot fluorescent microsphere chromatography and a preparation method and application method thereof. Background technique [0002] Tetanus is also known as ankylosis, and the common name is lock mouth. It is a specific infection in which Clostridium tetani invades the human body through skin or mucous membrane wounds, grows and reproduces in an anoxic environment, and when it enters the human body, it will act on the central nervous system to produce strong neurotoxins and cause muscle spasms. After the onset of the disease, most patients show muscle tension contraction and spasm, which makes the face appear "wry smile". It will also block the inhibition of the spinal cord to the sympathetic nerves, resulting in symptoms such as increased blood pressure, increased heart ra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/58G01N33/558G01N33/543G01N33/533
CPCG01N33/533G01N33/54346G01N33/558G01N33/56911G01N33/588
Inventor 李朝辉陈娟孟红敏杨冉屈凌波
Owner ZHENGZHOU UNIV
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