96-hole ELISA (enzyme linked immunosorbent assay) plate-immobilized influenza virus imprint fluorescent sensor and application

A fluorescent sensor and influenza virus technology, applied in instruments, scientific instruments, measuring devices, etc., can solve the problems of molecular imprinting technology, which is rarely reported, and achieve the effect of meeting the recovery rate, improving sensitivity, and overcoming limitations

Inactive Publication Date: 2018-06-26
MINJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

When the target molecular size is further enlarged to virus particles, new difficulties and challenges wil

Method used

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  • 96-hole ELISA (enzyme linked immunosorbent assay) plate-immobilized influenza virus imprint fluorescent sensor and application
  • 96-hole ELISA (enzyme linked immunosorbent assay) plate-immobilized influenza virus imprint fluorescent sensor and application
  • 96-hole ELISA (enzyme linked immunosorbent assay) plate-immobilized influenza virus imprint fluorescent sensor and application

Examples

Experimental program
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Embodiment 1

[0031] A 96-well microtiter plate immobilized influenza virus imprinted fluorescent sensor, the specific steps are as follows:

[0032] 1) Immobilization of template molecules: Add 30 μL of 50 μg / mL influenza virus H1N1 to each well of a 96-well ELISA plate, incubate at room temperature for 1 hour, and wash away unbound virus particles with ultrapure water;

[0033] 2) Preparation of polymer film: the molar ratio of aniline and APS is 3:1, pre-polymerized in the reagent tube, add 50 μL to each well, incubate for 30 min, wash away unpolymerized reactants with ultrapure water, 200 μL 1 mol / Incubate with L NaCl for 90 min, wash away template virus, and wash with ultrapure water to remove residual eluent;

[0034] 3) Recognition of the target virus on the imprinted membrane: take 30 μL of 0.01-1.0 μg / mL influenza virus H1N1, incubate for 60 min, and wash away the unadsorbed target virus with ultrapure water;

[0035] 4) HRP is labeled on the surface of influenza virus immobilize...

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Abstract

The invention relates to a 96-hole ELISA (enzyme linked immunosorbent assay) plate-immobilized influenza virus imprint fluorescent sensor and application, and belongs to the technical field of biological molecular detection. The 96-hole ELISA plate-immobilized influenza virus imprint fluorescent sensor is specifically characterized in that an influenza virus particle is used as a template moleculeand is immobilized onto a 96-hole ELISA plate, a thin layer of polyaniline is polymerized to imprint the template virus, the template molecule is eluted, and the left two-dimensional imprinting cavity can be used for specifically identifying the target virus; HRP (horse raddish peroxidase) is used as a biological marker and is directly marked onto the surface of the influenza virus immobilized onto an imprinting film, the HRP can effectively catalyze a H2O2-OPDA (hydrogen peroxide and o-phenylenediamine) system to produce sensitive fluorescent effect, and the produced fluorescent intensity has direct relationship with the concentration of the influenza virus, so as to build a novel influenza virus fluorescent sensor; the prepared imprinting polymer film has the characteristic of an artificial antibody, is used for identifying and detecting the influenza virus, and has been successfully applied to detect a serum sample. The 96-hole ELISA plate-immobilized influenza virus imprint fluorescent sensor provides an influenza virus identifying and detection method with high stability, high specificity and high sensitivity.

Description

technical field [0001] The invention relates to a 96-well microplate plate-immobilized influenza virus imprinted fluorescent sensor and its application, belonging to the technical field of biomolecular detection. Background technique [0002] Influenza virus, referred to as influenza virus, belongs to the Orthomyxoviridae family and is a single-stranded negative-sense RNA virus. It can cause acute upper respiratory tract infection, and spread rapidly through the air through breathing, speech, etc., and spread all over the world. There are often periodic pandemics. The influenza pandemic caused by the new subtype influenza virus threatens human health. With the globalization of human economy and culture, suitable conditions have been created for the global influenza pandemic. In order to realize the detection and prevention of the incubation period of influenza virus transmission, some simple instruments and rapid and low-cost detection methods have received great attention....

Claims

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Application Information

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IPC IPC(8): G01N33/569
Inventor 李艳霞郑黎云陈毅挺黄露娄本勇
Owner MINJIANG UNIV
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