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Alpha1-antitrypsin immunochromatographic detection reagent card and preparation method thereof

An immunochromatographic detection and antitrypsin technology, which is applied in the field of biomedicine, can solve the problems of inapplicable diagnosis of emergency patients and clinical patients, cumbersome operation, and long time.

Inactive Publication Date: 2018-12-04
广州华澳生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the main methods for detecting α1-antitrypsin include: serum trypsin inhibition capacity test, one-way immunodiffusion method, rocket immunoelectrophoresis method and ELISA method, but all have the disadvantages of cumbersome operation and time-consuming, and are not suitable for emergency patients timely diagnosis of clinical patients

Method used

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  • Alpha1-antitrypsin immunochromatographic detection reagent card and preparation method thereof
  • Alpha1-antitrypsin immunochromatographic detection reagent card and preparation method thereof
  • Alpha1-antitrypsin immunochromatographic detection reagent card and preparation method thereof

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Experimental program
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Effect test

preparation example Construction

[0058] The steps of the preparation method of the coating film are:

[0059] ① Dilute goat anti-rabbit or goat anti-mouse antibody with 0.02M phosphate buffer solution to 0.5~2mg / mL, add 3% methanol, it is the working solution of the quality control area;

[0060] ② Dilute the anti-α1-antitrypsin antibody with 0.02M phosphate buffer solution to 0.8-1.5 mg / mL, add 3% methanol, it is the working solution of the detection area;

[0061] ③ Coat the solution obtained in step ① and step ② on the nitrocellulose membrane at 0.8-1.2 μL / cm with a gold-spraying device. The distance between the detection area and the quality control area is 0.4-0.7 cm. In the middle position, put the nitrocellulose membrane in a 37°C oven and let it dry.

[0062] The preparation method of the sample pad is to soak the sample pad with a sample pad buffer, and the sample pad buffer contains 0.5-5wt% BSA, 0.5-5wt% polyvinylpyrrolidone, 0.5-5wt% polyethylene glycol, 0.1-5wt% Tween-20, 1-5wt% Triton-100 in 0...

Embodiment 1

[0063] Example 1, preparation method of α1-antitrypsin immunochromatographic detection reagent card (labeled with quantum dots or rare earth complexes):

[0064] 1) The preparation method of the sample pad is:

[0065] Use 0.01M phosphoric acid containing 0.5~5wt% BSA, 0.5~5wt% polyvinylpyrrolidone, 0.5~5wt% polyethylene glycol, 0.1~5wt% Tween-20, 1~5wt% Triton-100 for the sample pad After soaking in salt buffer solution (0.01 M disodium hydrogen phosphate: 0.01M sodium dihydrogen phosphate = 81:19 (V: V)) (pH 7.4), dry at 37°C or in an environment with a relative humidity of ≤35% , cropped for use.

[0066] The sample pad buffer is preferably 0.5wt% BSA, 0.5wt% polyvinylpyrrolidone, 0.5wt% polyethylene glycol, 0.25wt% Tween-20 and 1wt% Triton-100 dissolved in 0.01M phosphate buffer.

[0067] 2) The preparation method of anti-α1-antitrypsin antibody quantum dots or rare earth complex labeling pad is:

[0068] ① Dissolve and dilute the water-soluble carboxyl CdSe / ZnS or Eu(T...

Embodiment 2

[0088] Example 2, preparation method of α1-antitrypsin immunochromatographic detection reagent card (labeled with fluorescent nanospheres or colored nanospheres):

[0089] 1) Preparation of anti-α1-antitrypsin antibody polystyrene nanofluorescent microspheres or colored polystyrene microspheres labeling pad:

[0090] ① Use 0.1M MES buffer solution with pH 6.0 (with 0.1M MES aqueous solution, adjust the pH with 1 M NaOH solution) to mix carboxypolystyrene nano fluorescent microspheres (particle size 190nm) or colored polystyrene microspheres (particle size 200nm) ) to a solid content of about 0.2%;

[0091] ② Take 1mL polystyrene nanofluorescent microspheres or colored polystyrene microspheres with a solid content of 0.2%, add 50μL EDC solution (10mg / mL) and 50μL sulfo-NHS solution (50mg / mL), activate for 10min, 16500r / min Centrifuge for 30 minutes, remove the supernatant, add 300 μg anti-α1-antitrypsin antibody (primary antibody), and mix the coupling reaction for 1 hour;

...

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Abstract

The invention discloses an alpha1-antitrypsin immunochromatographic detection reagent card and a preparation method thereof, and aims at providing an alpha1-antitrypsin detection method with the advantages of fastness, simplicity, high sensitivity and reliable detection result. The alpha1-antitrypsin immunochromatographic detection reagent card comprises a box body, the box body is provided with abottom plate and a box lid, the box lid is provided with a sample adding hole and a result detecting window, a detection test paper is arranged in the box body, the bottom plate is arranged under thetest paper, the test paper is provided with a detection area, a sample pad, an anti-alpha1-antitrypsin antibody tracer marked pad, a coated film and an absorbent pad are arranged in the detection area, the coated film is provided with an alpha1-antitrypsin antibody detection area T line and a quality control area C line, and the T line and the C line are arranged in parallel. The detection reagent card and the detection method have the advantages of high sensitivity, high specificity, rapidness, simplicity, and realization of quantitative or qualitative detection of an analyte.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to a detection reagent card for measuring the content of α1-antitrypsin in blood (serum, plasma, whole blood) and a preparation method thereof. Background technique [0002] α1-antitrypsin (AAT), also known as α1 trypsin inhibitor, is a member of the serine protease inhibitor family. It is a single-chain glycoprotein consisting of a polypeptide chain connected with 3 oligosaccharide chains, with a molecular weight of about 52kDa, PI The value is 4.8. As the most important protease inhibitor in the human body, α1-antitrypsin is mainly synthesized by liver cells, and can inhibit more than 90% of the protease activity in normal plasma. It can mainly inhibit neutrophil elastase, in all serine Among protease inhibitors, α1-antitrypsin has strong vascular permeability, so α1-antitrypsin often has a high concentration in the local inflammation, and it is more specific to elastase, ma...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/58G01N33/558G01N33/533G01N33/573
CPCG01N33/533G01N33/558G01N33/573G01N33/582G01N33/587G01N33/588G01N33/68G01N2800/7095
Inventor 于龙波于永涛黎权刘日俊
Owner 广州华澳生物科技有限公司
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