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

An immunochromatographic test strip and Mycoplasma pneumoniae technology, applied in the field of medical immunodetection, can solve the problems of low sensitivity and low accuracy, and achieve the effects of good luminescence stability, narrow emission peak and symmetrical peak shape

Active Publication Date: 2014-01-22
北京华卫天和生物科技有限公司
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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 pneumonic mycoplasma, quantum dot-labeled immunochromatographic test paper and preparation method thereof
  • Method for detecting pneumonic mycoplasma, quantum dot-labeled immunochromatographic test paper and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: A kind of quantum dot-labeled immunochromatographic test paper is provided with plastic plate, nitrocellulose membrane, glass cellulose membrane A, quantum dot-labeled glass cellulose membrane B of mycoplasma pneumoniae IgM monoclonal antibody, absorbent paper, so The above-mentioned glass cellulose film A is the glass cellulose film purchased on the market without sample;

[0033] Wherein, the plastic plate is pasted with glass cellulose membrane A, quantum dot-labeled glass cellulose membrane B of mycoplasma pneumoniae IgM monoclonal antibody, nitrocellulose membrane, and absorbent paper in sequence;

[0034] Wherein, one end of the nitrocellulose membrane has a Mycoplasma pneumoniae 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 Mycoplasma pneumoniae IgM monoclonal antibody is located at the other end of the glass cellulose membrane B, corr...

Embodiment 2

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

[0042] (1) Coupling of quantum dots and Mycoplasma pneumoniae IgM monoclonal antibody:

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

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

[0045] Add 150-200 uL of Mycoplasma pneumoniae IgM monoclonal antibody;

[0046] Shaker reaction for 1 to 4 hours;

[0047] 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 the Mycoplasma pneumoniae polyclonal antibody and rabbit anti-mouse secondary antibody with 0.05-0.15M PBS buffer, spray 0.5g / L Mycoplasma pneumoniae polyclonal antibody and 1.0g / L rabbit a...

Embodiment 3

[0058]Embodiment 3: detect Mycoplasma pneumoniae IgM monoclonal antibody with described test paper, comprise the following steps: Sample spot is approached on the assembled test paper one end of Mycoplasma pneumoniae IgM monoclonal antibody, after reacting 5min, in the ultraviolet analyzer Observation results. PBS buffer solution and normal human blood were used as blank controls.

[0059] Result judgment: under the premise that the C band shows a red fluorescent band, the intensity of the fluorescent band of the T band is visually compared with the blank. The weaker the fluorescence, the lower the concentration of the tested substance in the test solution.

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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 pneumonic mycoplasma by adopting an immunological method. According to the quantum dot-labeled immunochromatographic test paper, a glass cellulose membrane A, a quantum dot-labeled pneumonic mycoplasma IgM 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 pneumonic mycoplasma 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 pneumonic mycoplasma IgM monoclonal antibody is located at the other end of the glass cellulose membrane B and is corresponding to the inspection strip T and the quality control strip C, and the quantum dot-labeled pneumonic mycoplasma IgM 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 inspection method by about 1000 times.

Description

technical field [0001] The invention relates to a medical immunological detection method, in particular to a method for detecting mycoplasma pneumoniae in an immunological method by using quantum dot-labeled immunochromatographic test paper. Background technique [0002] Mycoplasma pneumoniae (Mycoplasma pneumoniae, MP) is a kind of microorganism between viruses and bacteria, and it is a kind of smallest prokaryotic microorganism that can grow and reproduce on non-living medium. MP infection can occur at any age, especially between 5 and 20 years old. MP spreads through droplets in the form of aerosol particles and causes Mycoplasma pneumoniae pneumonia (MPP) after infection. Regional epidemics occur every 3 to 5 years, accounting for 10% to 20% of all types of pneumonia. 30% to 50% of recruits suffering from pneumonia are caused by Mycoplasma pneumoniae; accounting for more than 1 / 3 of non-bacterial pneumonia. In addition, it can also cause changes in various extrapulmona...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/531
CPCG01N33/56933
Inventor 文德敏申有长于晓永
Owner 北京华卫天和生物科技有限公司
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