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SARS-CoV-2 neutralizing antibody fluorescence immunochromatography detection test strip and preparation method thereof

A fluorescent immunochromatography and detection test paper technology, applied in immunoassays, biological tests, fluorescence/phosphorescence, etc., can solve cumbersome and time-consuming problems, and achieve the effects of fast detection, no professional operation, and low cost.

Pending Publication Date: 2021-10-19
广州中医药大学顺德医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The virus neutralization test (cVNT) can detect neutralizing antibodies in the patient's blood, but it needs to be tested in a third-level biosafety laboratory, which is cumbersome and time-consuming

Method used

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  • SARS-CoV-2 neutralizing antibody fluorescence immunochromatography detection test strip and preparation method thereof
  • SARS-CoV-2 neutralizing antibody fluorescence immunochromatography detection test strip and preparation method thereof
  • SARS-CoV-2 neutralizing antibody fluorescence immunochromatography detection test strip and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1 A new crown neutralizing antibody fluorescent immunochromatography detection test strip and its preparation method

[0031] 1. Experimental method

[0032] 1. Preparation of sample pad

[0033] Soak the glass fiber with a specification of 25cm×30cm in the buffer solution of the sample pad, take it out after soaking for 3 to 5 minutes, dry it at room temperature for 16 to 18 hours, and cut it into a sample pad with a size of 2cm×30cm.

[0034] The buffer formula of the sample pad is as follows: 0.5% (mass fraction) BSA, 2.5% (mass fraction) sucrose, 1.5% (volume fraction) Tween-20 are dissolved in 0.015M pH9 PB buffer.

[0035] 2. Preparation of fluorescent markers and binding pads

[0036] 1) Preparation of fluorescently labeled ACE2: Add 200 μL of europium-labeled time-resolved fluorescent microspheres into a 2 mL EP tube, then add 800 μL of 0.1M pH6.0 MES, and mix well in the dark; then add 200 μL of 1 mg / mL NHS (N -Hydroxysuccinimide) and 200 μL of 50...

Embodiment 2

[0066] Example 2 Detection of Blood Samples of New Coronary Rehabilitation Patients and Vaccine Injectors

[0067] 1. Experimental method

[0068] Clinical samples were detected using the test strip of Example 1. In this example, 8 recovered patients, 8 vaccinators, and 16 healthy individuals were included as controls. To evaluate the detection effect of the test strip of Example 1 on neutralizing antibodies against the new coronavirus.

[0069] 2. Experimental results

[0070] The test results are shown in Table 1. The inhibition rates of 8 recovered patients and 8 vaccinated patients were all greater than 30%. This test strip can detect neutralizing antibodies in serum.

[0071] Table 1:

[0072]

[0073]

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Abstract

The invention discloses a SARS-CoV-2 neutralizing antibody fluorescence immunochromatography detection test strip. The test strip comprises a sample pad, a combination pad, a reaction film and an absorption pad which are sequentially jointed on a bottom plate, wherein the combination pad is coated with ACE2 coupled with time-resolved fluorescent microspheres and goat anti-chicken IgG; the reaction film is provided with a detection line and a quality control line, the detection line is coated with RBD, and the quality control line is coated with a chicken IgY antibody. The SARS-CoV-2 neutralizing antibody fluorescence immunochromatography detection test strip provided by the invention has the characteristics of low cost, rapid, sensitive and instant detection, no need of professional operation and the like, and can be completed only in a secondary biosafety laboratory. The test strip can effectively monitor the vaccination reaction in clinical research and serve as an evaluation index of the later vaccine effect; the detection rate of SARS-CoV-2 is increased, and missing detection caused by false negative of nucleic acid detection is made up; and detecting the content of neutralizing antibodies in the body of the SARS-CoV-2 rehabilitation patient so as to judge whether rework and reproduction can be performed or not and whether a reinfection risk exists or not.

Description

technical field [0001] The invention relates to the technical field of virus detection, in particular to a novel coronavirus neutralizing antibody fluorescent immunochromatographic detection test strip and a preparation method thereof. Background technique [0002] The new coronavirus (SARS-CoV-2) is a new type of β-coronavirus (β-coronavirus), with a diameter of 60-140nm and a crown-like shape under an electron microscope. The spike protein (receptorbinding domain, RBD) of SARS-CoV-2 can bind human ACE2 protein, so that the virus enters cells and replicates. At the same time, RBD is also the main target of neutralizing antibodies. The new coronavirus neutralizing antibody (NAb) is an antibody produced in patients with new coronavirus pneumonia and new crown vaccine injectors. It can bind to the RBD of the S protein to block the combination of the virus and human ACE2, showing direct neutralizing activity. Therefore, the detection of neutralizing antibodies can determine w...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/558G01N33/58G01N33/543G01N33/52G01N21/64
CPCG01N33/56983G01N33/558G01N33/582G01N33/54346G01N33/52G01N21/6408G01N21/6428G01N2469/20G01N2333/165
Inventor 徐建华张嘉琪张晓丽李宜哲徐涛滕祥云何金勇
Owner 广州中医药大学顺德医院