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A kind of new coronavirus antibody detection fluorescent reagent and preparation method

A fluorescent reagent and antibody detection technology, applied in biological testing, measuring devices, immunoassays, etc., can solve the problems of insufficient automation, low sensitivity, and cumbersome steps, and achieve the effect of reducing research and development costs, simplifying test methods, and cumbersome detection steps.

Active Publication Date: 2022-05-17
HUAZHONG UNIV OF SCI & TECH +1
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Problems solved by technology

[0006] In view of the low sensitivity, cumbersome steps, and insufficient automation of various existing methods for detecting antibodies based on immune reactions, the purpose of the present invention is to provide a fluorescent reagent for detecting antibodies against the new coronavirus and a preparation method. The surface of quantum dots is marked, and the characteristics of easy synthesis and regulation of the two and their size matching are used to improve the marking effect, so that the antigen-antibody binding immune reaction on the surface of quantum dots can cause a significant change in the peak fluorescence wavelength or intensity of quantum dots. Using a fluorometer or The spectrometer is used for automatic detection to realize the detection and quantitative analysis of highly sensitive, highly specific, and high-throughput novel coronavirus antibodies

Method used

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  • A kind of new coronavirus antibody detection fluorescent reagent and preparation method
  • A kind of new coronavirus antibody detection fluorescent reagent and preparation method
  • A kind of new coronavirus antibody detection fluorescent reagent and preparation method

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Embodiment 1

[0037] (1) Synthesis of CdSe / ZnS core-shell quantum dots under visible light. Specifically, it is divided into (a) preparation of CdSe quantum dots and (b) coating of ZnS shells. (a) Add 0.165g of CdO and 0.28g of stearic acid into a three-necked flask, stir and heat to 150°C and maintain a vacuum state for 30min; after cooling down to 50°C, add 2.43g of tri-n-octylphosphine oxide (TOPO) and 2.43g of ten Hexamine (HAD), heated to 250°C again and maintained; 0.1g of selenium powder was dissolved in 0.54g of tri-n-octylphosphine (TOP) and 2.1g of dioctylamine and injected into the flask, and kept at 200°C for reaction Until the solution is colorless, it is cooled to room temperature to complete the preparation of CdSe quantum dots. (b) Weigh 0.241g of zinc acetate, 1.06g of TOPO and 1.06g of HDA in a container, stir and heat to 200°C, and keep it under vacuum for 30 minutes; use a syringe to take the mixed solution and add it evenly to a three-necked CdSe quantum dot flask, and...

Embodiment 2

[0043] (1) Synthesis of CdTe quantum dots under visible light. Specifically, 0.05 mL of 100 mmol / L CdCl 2 Add to the Erlenmeyer flask, then add 5.25mL of water, 0.25mL of 40mmol / L mercaptopropionic acid MPA, 0.25mL of 4mmol / L Na 2 TeO 3 , 0.6mg of NaBH 4 and 34.2ml of 85% N 2 h 4 ·H 2 O. Magnetic stirring for 2 h ensures the growth of quantum dots. The quantum dots can be precipitated by adding 2-propanol to the QD solution and centrifuging, and the precipitates are collected after repeated washing several times, and the CdTe quantum dots with orange-red light emission are obtained after vacuum drying.

[0044] (2) On the surface of the quantum dots, a new coronavirus-specific B-cell epitope antigen polypeptide whose sequence is shown in SEQ ID No.1, which can specifically detect the new coronavirus antibody, is introduced to replace or label the surface ligands of the quantum dots. Specifically, choose 1 mL of phosphate buffer, add 2 μL of 10 mg / mL CdTe quantum dots s...

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Abstract

The invention belongs to the technical field of virus antibody detection, and discloses a novel coronavirus antibody detection fluorescent reagent and a preparation method, wherein the preparation method comprises the following steps: (1) synthesizing visible light fluorescent colloidal quantum dots; (2) synthesizing visible light fluorescent colloidal quantum dots The pre-prepared SARS-CoV-2 antigenic peptides are labeled on the surface to obtain quantum dot fluorescent reagents. The present invention is based on the change of peak fluorescence wavelength or intensity of quantum dot fluorescence caused by antigen-antibody binding immune reaction, which can be detected automatically by using a fluorometer or a spectrometer to achieve highly sensitive, highly specific, and high-throughput detection and quantitative analysis of antibodies against the novel coronavirus.

Description

technical field [0001] The invention belongs to the technical field of virus antibody detection, and more specifically relates to a novel coronavirus antibody detection fluorescent reagent and a preparation method. Background technique [0002] Rapid and accurate virus detection methods are of great significance for early detection, early isolation, early diagnosis, and early treatment of virus-infected people. Taking the COVID-19 novel coronavirus as an example, due to its characteristics of rapid reproduction, long incubation period, and high infectivity, it has caused large-scale crowd infection events around the world, causing serious harm to personal safety and social stability. Therefore, the development of rapid detection methods for COVID-19 antibodies and the rapid and accurate diagnosis of virus-carrying patients are very important for disease prevention and treatment. [0003] Colloidal gold kits / test strips, as one of the mainstream methods for rapid virus detec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/569G01N33/58G01N33/533
CPCG01N33/6854G01N33/56983G01N33/582G01N33/588G01N33/533G01N2333/165G01N2469/20Y02A50/30
Inventor 刘欢徐可李华曜翟博慧龙文博李龙胡志响陈浩
Owner HUAZHONG UNIV OF SCI & TECH
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