Method for detecting rubella virus, quantum dot-labeled immunochromatographic test paper and preparation method thereof
An immunochromatographic test strip and rubella virus technology, applied in the field of medical immunodetection, can solve the problems of low accuracy and low sensitivity, and achieve the effects of narrow emission peak, good luminescence stability and symmetrical peak shape.
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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 rubella virus IgG monoclonal antibody glass cellulose membrane B, absorbent paper, so The above-mentioned glass cellulose film A is the glass cellulose film purchased on the market without sample;
[0033] Wherein, glass cellulose membrane A, glass cellulose membrane B of quantum dot-labeled rubella virus IgG monoclonal antibody, nitrocellulose membrane, and absorbent paper are pasted sequentially on the plastic plate;
[0034] Wherein, there are rubella virus polyclonal antibody and rabbit anti-mouse secondary antibody at one end of the nitrocellulose membrane, so as to form detection zone T and quality control zone C;
[0035] Wherein, the rubella virus IgG monoclonal antibody labeled with quantum dots is located at the other end of the glass cellulose membrane B, corresponding to the dete...
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 rubella virus IgG 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 rubella virus IgG monoclonal antibody 150-200uL;
[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 rubella virus polyclonal antibody and rabbit anti-mouse secondary antibody with 0.05-0.15M PBS buffer, spray 0.5g / L rubella virus polyclonal antibody and 1.0g / L rabbit anti-mouse secondary antibody on one end ...
Embodiment 3
[0058]Embodiment 3: detect rubella virus with described test paper, comprise the following steps: spot sample on the assembled test paper close to one end of rubella virus IgG monoclonal antibody, after reacting for 5min, observe the result in the ultraviolet analyzer . 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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