Rapid detection method for virus activity

A detection method and virus technology, applied in the biological field, can solve the problem of few reports on mammalian cells, and achieve the effect of improving the imprinting effect and ensuring the structural stability.

Active Publication Date: 2020-02-07
武汉南嘉木实业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to develop a rapid detection technology for virus activity without culturing, the present invention uses molecularly imprinted polymers (molecularly imprinted polymers, MIPs). At present, there have been a small amount of research on the preparation of bacterial MIPs, and there are few reports on the MIPs of mammalian cells.

Method used

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  • Rapid detection method for virus activity
  • Rapid detection method for virus activity
  • Rapid detection method for virus activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Preparation of fluorescent molecularly imprinted polymer I for Escherichia coli

[0059] The method for preparing the fluorescent molecularly imprinted polymer I of Escherichia coli comprises the steps of:

[0060] 1) Preparation of reagents:

[0061] Dopamine solution Ⅰ: ammonium persulfate and dopamine are dissolved in 10mmol / L Tris-HCl buffer solution (pH=8 of Tris-HCl buffer solution) at a molar ratio of 2:1;

[0062] Dopamine solution II: dopamine is dissolved in 10mmol / L Tris-HCl buffer solution (Tris-HCl buffer solution pH=8);

[0063] Dopamine / dansyl dopamine mixed solution: dopamine: dansyl dopamine: ammonium persulfate molar ratio=4:1:1 molar ratio dissolved in 10mmol / L Tris-HCl buffer solution (Tris-HCl buffer solution pH=8) ;

[0064] Carrier material: 96-well microtiter plate is used as the carrier material;

[0065] Host target cells: 10 4 ~10 6 CFU / mL of Escherichia coli liquid, Escherichia coli using E. Coli 285;

[0066] Eluent: deioniz...

Embodiment 2

[0073] Example 2 Preparation of fluorescent molecularly imprinted polymer II for Escherichia coli

[0074] Compared with Example 2, the preparation method of Example 2 differs in that Example 2 does not carry out step 2) in Example 1, that is, Example 2 does not use the polydopamine-modified dopamine-carrier complex as a carrier, and It is to directly polymerize dopamine, dansyl dopamine, and Escherichia coli on the carrier material. The specific process is as follows:

[0075] 1) Preparation of Escherichia coli-dansyl dopamine / dopamine-carrier complex: add dopamine / dansyl dopamine mixed solution and host target cells (E. at 37°C with a rotational speed of 150rpm) and slowly shake for 72 hours, then pour out the liquid in the hole to obtain the Escherichia coli-dansyl dopamine / dopamine-carrier complex;

[0076] 2) Elution of host target cells: wash the Escherichia coli-dansyl dopamine / dopamine-carrier complex repeatedly with an eluent, and elute the Escherichia coli in the E...

Embodiment 3

[0077] Example 3 Preparation of fluorescent molecularly imprinted polymer III for Escherichia coli

[0078] The preparation method of the fluorescent molecularly imprinted polymer III of Escherichia coli is the same as that of Example 1, except that in this example, the dopamine solution I in step 2) of Example 1 is changed to dopamine solution II, and the dopamine solution I and dopamine solution The difference of II is whether ammonium persulfate is added or not. The method for preparing the fluorescent molecularly imprinted polymer III of Escherichia coli in this example will not be repeated here.

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Abstract

The invention relates to the biotechnical field, and especially relates to a method for rapidly detecting the activities of a virus. The method for rapidly detecting the activities of the virus comprises the following steps: 1) preparing a dopamine / dansyl dopamine mixed solution, uniformly mixing the dopamine / dansyl dopamine mixed solution, a host target cell and a carrier, carrying out a polymerization reaction on the surface of the carrier to obtain a host target cell- dansyl dopamine / dopamine-carrier complex, and finally eluting the host target cell from the complex by using an eluent to obtain a fluorescent molecularly-imprinted polymer; and 2) adding the host target cell and a virus sample to the fluorescent molecularly-imprinted polymer, instantly measuring the fluorescence value I0, measuring the fluorescence value I after the virus in the virus sample completely invades the host target cell, allowing the value of (I / I0 - 1) to indicate the fluorescence change level, and determining whether the virus sample contains the target virus or not and the content of the target virus according to the fluorescence change level. The detection method is fast, safe and accurate.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a rapid detection method for virus activity. Background technique [0002] Many human diseases are caused by viruses. At present, there are few effective antiviral drugs. The prevention and treatment of viral diseases mainly rely on vaccines and other preventions. Therefore, to control the spread of the virus and minimize the mortality caused by the virus, rapid and accurate detection of virus activity is essential. [0003] The virus needs to proliferate in the host cell. The live virus first forms non-specific adsorption with the host cell through random collision and electrostatic attraction; then the virus binds to the specific adhesion receptor on the surface of the host cell through its own adhesin, causing the cell membrane Altered, viral nucleic acid or virus enters target cells for biosynthesis, assembly and release. The traditional virus detection method is to determine t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 吕斌刘燕婕李宁叶磊吴中乔邓耘项阳
Owner 武汉南嘉木实业有限公司
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