Method for detecting SARS-Cov-2 by using blood glucose meter

A coronavirus, blood glucose meter technology, applied in microorganism-based methods, biochemical equipment and methods, instruments, etc., can solve the problems of high cost, long time, inability to efficiently process multiple samples, etc. low cost effect

Pending Publication Date: 2020-07-17
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
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Problems solved by technology

The detection methods for viruses include electrophoresis, radioactive analysis, traditional fluorescence spectroscopy, immunoassay, chromatography, etc. In recent years, PCR-based molecular biology detection methods have become very important in virus detection. Methods, in the diagnosis of the new coronavirus, the detection method based on PCR technology is also mainly used. Although the PCR method is more acc

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  • Method for detecting SARS-Cov-2 by using blood glucose meter

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preparation example Construction

[0044] The most preferred detailed process for preparing magnetic beads-nucleic acid probe 1 is:

[0045] First, activate the streptavidin-coated magnetic beads. The activation of streptavidin-magnetic beads in the embodiment of the present invention specifically includes the following steps: place the magnetic bead bottle on a vortex shaker for 20 s, oscillate to resuspend the magnetic beads . Pipette 100L of magnetic beads into a new centrifuge tube. Place the centrifuge tube on the magnetic separator and let it stand for 1 min (this operation will be referred to as magnetic separation later), suck off the supernatant with a pipette, and remove the centrifuge tube from the magnetic separator. Add 1mL Buffer I (Buffer 1 solution is 0.01MPBST solution, the pH of Buffer 1 solution is 7.2, and Buffer 1 solution contains 0.2% Tween-20) into the centrifuge tube, cover the centrifuge tube cap, and vortex fully to resuspend the magnetic beads. Magnetically separate and remove the ...

Embodiment 1

[0064] 1. Connect 60 μL of streptavidin-magnetic beads to nucleic acid probe 1 to obtain streptavidin-magnetic beads connected to nucleic acid probe 1, then perform magnetic separation and wash 3 times with Buffer A solution (magnetic stand Washing by magnetic adsorption, each time for 2 minutes), is magnetic bead-nucleic acid probe 1.

[0065] 2. Take 400μL 2mg / mL sucrase (dissolved in BufferA solution) and add 1mg sulfo-SMCC coupling agent, mix well at room temperature (under the condition of rotating speed of 20-30r / min), pH is 6.5-7.5, shake for 1h , centrifuged to remove insoluble sulfo-SMCC, and then ultrafiltered 8 times with BufferA solution to obtain the coupling agent-sucrase complex.

[0066] 3. Take 30 μL of 100 μM sulfhydryl-modified nucleic acid probe 2 (dissolved in ultrapure water), add 10 μL of 0.2M PB (pH5.5) buffer solution and 2 μL of 50 mM TCEP (dissolved in ultrapure water), mix well, and place at room temperature for 1 hour , was ultrafiltered 8 times w...

Embodiment 2

[0078] A kit for detecting novel coronavirus using a blood glucose meter to detect the nucleic acid sequence of ORF1ab region of novel coronavirus, comprising the following components, reagents for capturing viral target nucleic acid sequences, viral signal conversion reagents and auxiliary reagents; the capture virus The reagent of target nucleic acid sequence comprises the nucleic acid probe 2 that is connected with magnetic bead-nucleic acid probe 1, is connected with sucrase; The signal conversion reagent of described virus comprises sucrose solution; Described auxiliary reagent is used in virus detection process Reagent comprises Buffer A and morpholineethanesulfonic acid-Tween 20 solution; Described Buffer A solution is pH 7.2, contains the PBST solution of 0.5% Tween-20 and 1g / L BSA; Described morpholineethanesulfonic acid - The formula of the Tween 20 solution is: 100 mM morpholinoethanesulfonic acid, 0.05% Tween 20, pH 5.0. The concentration of the sucrose solution is 0...

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Abstract

The invention discloses a method for detecting SARS-Cov-2 by using a blood glucose meter. The method includes the following steps: (1) designing a nucleic acid probe 1 and a nucleic acid probe 2 according to a target nucleic acid sequence of the SARS-Cov-2, and preparing magnetic bead-nucleic acid probe 1 and invertase-nucleic acid probe 2; (2) mixing the magnetic bead-nucleic acid probe 1 and theinvertase-nucleic acid probe 2 with a sample to be tested to obtain a mixture, wherein the mixture is used to jointly capture the target nucleic acid sequence in the sample to be tested; (3) converting the signal of the target nucleic acid sequence captured in the step (2) into a glucose signal to obtain a mixed solution to be tested; and (4) detecting the converted glucose signal in the mixed solution to be tested by using the blood glucose meter to obtain the amount of the SARS-Cov-2 in the sample to be tested. The method provided by the invention detects the amount of glucose through the blood glucose meter to obtain the amount of a biomarker, can realize the purpose of simple and rapid detection, and can obtain quantitative detection results, the results can be digitally output by theblood glucose meter, and the method is short in consumed time and low in cost.

Description

technical field [0001] The invention relates to the field of biomolecular detection of viruses, and relates to a method for detecting novel coronaviruses by utilizing the recognition of glucose by a blood glucose meter and the characteristic of decomposing sucrose by sucrase. Background technique [0002] Novel coronavirus pneumonia (COVID-19) is an acute respiratory infectious disease caused by 2019 novel coronavirus (SARS-Cov-2) infection. The infection rate is high and the epidemic situation is serious. The detection and diagnosis of the new coronavirus is an important part of effective control of the epidemic. Efficient virus detection technology has also become a hot spot in research and development. The detection methods for viruses include electrophoresis, radioactive analysis, traditional fluorescence spectroscopy, immunoassay, chromatography, etc. In recent years, PCR-based molecular biology detection methods have become very important in virus detection. Methods, ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6834C12Q1/54C12Q1/34G01N33/66C12R1/93
CPCC12Q1/701C12Q1/6834C12Q1/54C12Q1/34G01N33/66G01N2333/924C12Q2565/519C12Q2563/143C12Q2563/149
Inventor 刘斐王彬单衍可陆雨楠李嘉豪
Owner NANJING AGRICULTURAL UNIVERSITY
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