Detection kit for visibly detecting virus and virus detection method

A virus and biotinylation technology, applied in the field of analysis and detection, can solve the problems of complex instruments, long time-consuming, cumbersome detection steps, etc., and achieve the effects of high sensitivity, high sensitivity and strong specificity

Inactive Publication Date: 2014-04-09
WUHAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a simple, fast and highly sensitive method for visual detection of viruses, so as to overcome the problems of cumbersome detection steps, long time-consuming and complex instruments in the prior art

Method used

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  • Detection kit for visibly detecting virus and virus detection method
  • Detection kit for visibly detecting virus and virus detection method
  • Detection kit for visibly detecting virus and virus detection method

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Embodiment 1

[0030] The detection of embodiment 1 H9N2 avian influenza virus

[0031] The method of the present invention will be described in detail below by taking the H9N2 avian influenza virus as an example.

[0032] 1. Method

[0033] 1. Preparation of immunomagnetic spheres

[0034] Take 8 μL of superparamagnetic balls (Master Beads Carboxylic Acid 0215, 500 nm diameter) with carboxyl functional groups on the surface, and wash twice with 0.1 mol / L PBS at pH 6.0. Then the magnetic balls were dispersed in PBS (0.1 mol / L pH 6.0) solution containing 0.1 mol / L EDC and 0.1 mol / L NHS, and activated for 30 min. After washing three times with 0.1 mol / L pH 7.2 PBS, it was dispersed in 200 μL 0.1 mol / L pH 7.2 PBS solution, and then 10 μL 0.59 mg / mL monoclonal antibody solution against H9N2 surface HA was added and shaken. Incubate the reaction on bed for 4 h. After the reaction, wash three times with 0.1 mol / L PBS pH 7.2 to remove excess antibody, then disperse it in 200 μL 1% BSA, and ...

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Abstract

The invention relates to a detection kit for detecting virus and a virus detection method. An immune magnetic ball is used as a solid carrier, a biotinylation antibody and alkaline phosphatase (SA-ALP) labeled by a streptavidin marker are used as signal markers, a magnetic immune sandwich compound is formed on the surface of the immune magnetic ball, a substrate is catalyzed to be hydrolyzed by alkaline phosphatase (ALP) on the surface of the magnetic immune sandwich compound to produce a substance with strong reduction property, so that Ag+ is reduced to be AgO, and the AgO is deposited on the surface of nano gold to form Au/Ag nano particles, so that the target pathogene can be qualitatively and quantitatively detected by utilizing the color variation of colloid gold solution and the variation of an ultraviolet visible absorption spectroscopy. The invention also provides a detection kit for the method. According to the method, the magnetic immune analysis and the enzymatic metallization colorimetry are combined and the virus can be sensitively, rapidly and visibly detected.

Description

technical field [0001] The invention relates to a virus detection method, and also relates to a detection kit applied to the method, which belongs to the field of analysis and detection. Background technique [0002] The rapid detection of viruses is crucial to food safety, disease prevention and control, effective diagnosis and treatment, and control of the spread and infection of viruses. However, the current virus detection methods: such as virus isolation, serological detection methods, and ELISA are cumbersome, time-consuming, and low-sensitivity; PCR requires expensive instruments and sample pretreatment, so it is not suitable for rapid virus detection and identification. Immunomagnetic separation technology is a new type of immunoseparation technology developed with the progress of nanomaterials in recent years. Because magnetic beads have a large specific surface area, fast reaction kinetics, good separation and enrichment capabilities, and are easy to manipulate , ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/54326G01N33/56983G01N33/581G01N2333/11
Inventor 张志凌周川华赵静雅庞代文
Owner WUHAN UNIV
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