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Method for detecting poliovirus, quantum dot-labeled immunochromatographic test paper and preparation method thereof

An immunochromatographic test paper and polio technology, which is applied in the field of medical immunological detection, can solve the problems of low accuracy and low sensitivity, and achieve the effect of narrow emission peak, good luminescence stability, and symmetrical peak shape

Inactive Publication Date: 2014-01-22
BEIJING HUAWEI BRAVOBIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the commonly used detection method is the colloidal gold method. Although this method is fast, simple and easy to operate, it has low accuracy and low sensitivity.

Method used

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  • Method for detecting poliovirus, quantum dot-labeled immunochromatographic test paper and preparation method thereof
  • Method for detecting poliovirus, quantum dot-labeled immunochromatographic test paper and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: A quantum dot-labeled immunochromatographic test paper, comprising a plastic plate, a nitrocellulose membrane, a glass cellulose membrane A, a quantum dot-labeled glass cellulose membrane B for poliovirus IgG monoclonal antibody, and absorbent paper , the glass cellulose film A is a commercially available glass cellulose film without spotting;

[0033] Wherein, glass cellulose membrane A, glass cellulose membrane B marked with poliovirus IgG monoclonal antibody by quantum dots, nitrocellulose membrane and absorbent paper are pasted on the plastic plate in sequence;

[0034] Wherein, one end of the nitrocellulose membrane has a poliovirus polyclonal antibody and a rabbit anti-mouse secondary antibody to form a detection zone T and a quality control zone C;

[0035] Wherein, the quantum dot-labeled poliovirus IgG monoclonal antibody is located at the other end of the glass cellulose membrane B, corresponding to the detection zone T and the quality control zone ...

Embodiment 2

[0041] Embodiment 2: the preparation method of the test paper as above, as figure 1 shown, including the following steps:

[0042] (1) Conjugation of quantum dots with poliovirus IgG monoclonal antibody:

[0043] Take 100-200uL of 0.01M PBS buffer and 5-20uL of quantum dots with carboxyl groups attached to the surface;

[0044] Select a coupling reagent, and the coupling reagent is selected from hydroxythiosuccinimide, 1-(3-dimethylaminopropyl)-3ethylcarbodiamine hydrochloride;

[0045] Add 150-200uL of poliovirus IgG monoclonal antibody;

[0046] Shaker reaction for 1 to 4 hours;

[0047] Chromatography column filtration, centrifugal purification;

[0048] Block with 1% to 5% bovine serum albumin;

[0049] Store at 4°C;

[0050] (2) Preparation of test paper:

[0051] Dilute poliovirus polyclonal antibody and rabbit anti-mouse secondary antibody with 0.05-0.15M PBS buffer, spray 0.5g / L poliovirus polyclonal antibody and 1.0g / L rabbit anti-mouse secondary antibody on ni...

Embodiment 3

[0058] Example 3: Detecting poliovirus with the test paper, including the following steps: spotting the sample on the assembled test paper close to one end of the poliovirus IgG monoclonal antibody, reacting for 5 minutes, and placing the sample in an ultraviolet analyzer. Observation results. PBS buffer and normal human blood were used as blank controls.

[0059] Judgment of the results: On the premise that the C band shows a red fluorescent band, the intensity of the fluorescence band of the T band is visually observed. Compared with the blank control, the weaker the fluorescence, the lower the concentration of the tested substance in the solution to be tested.

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Abstract

The invention relates to a medimmune inspection method, and particularly relates to quantum dot-labeled immunochromatographic test paper, and a method for detecting poliovirus by adopting an immunological method. According to the quantum dot-labeled immunochromatographic test paper, a glass cellulose membrane A, a quantum dot-labeled poliovirus IgG monoclonal antibody glass cellulose membrane B, a cellulose nitrate membrane and absorbent paper are sequentially are bonded on a plastic board from bottom to top, wherein a poliovirus polyclonal antibody and a rabbit-anti-mouse secondary antibody are on one end of the cellulose nitrate membrane so as to form an inspection strip T and a quality control strip C; a quantum dot-labeled poliovirus IgG monoclonal antibody is located at one end of the glass cellulose membrane B and corresponds to the inspection strip T and the quality control strip C, and the quantum dot-labeled poliovirus IgG monoclonal antibody is located at one end of a sample feeding point. The inspection sensitivity of the method is higher than of the currently used method by about 1000 times.

Description

technical field [0001] The present invention relates to a medical immune detection method, in particular to a method for detecting poliovirus by immunological method using quantum dots to mark immunochromatographic test paper. Background technique [0002] Poliovirus (Poliovirus, or poliovirus) is the causative agent of polio (polio), also known as poliovirus. It is an unenveloped virus consisting of a single-stranded RNA and protein coat, about 25 nanometers in diameter, belonging to the genus Enterovirus of the Picornaviridae family. In addition to humans, monkeys can also be infected with this virus. Poliovirus is contracted through feces and diet and through contact. Hygiene measures have little effect on the route of infection. It first infects the digestive system and, in rare cases, spreads to the rest of the body through the bloodstream. In most cases, myelitis is asymptomatic, but sometimes it causes flu-like symptoms. In a small number of cases (about 1%) it i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/531
CPCG01N33/577G01N2333/105
Inventor 文德敏申有长于晓永
Owner BEIJING HUAWEI BRAVOBIO
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