Method for detecting treponema pallidum, quantum-dot labeled immunochromatography test paper and preparation method thereof

An immunochromatographic test strip and Treponema pallidum technology, applied in the field of medical immunodetection, can solve the problems of poor sensitivity and specificity, low accuracy, complicated operation and the like, and achieve the effects of symmetrical peak shape, narrow emission peak and good luminescence stability.

Active Publication Date: 2014-01-22
北京华卫骥生物医药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

RPR and TRUST are specific for Treponema pallidum, with poor sensitivity and specificity (up to 30% of false positives and false negatives); although the colloidal gold method is fast, simple and easy to operate, it has low accuracy and low sensitivity ; while TPPA has high sensitivity and specificity, but the source of the natural spirochete antigen used is difficult, expensive, complicated to operate, and the judgment result is subject to subjective factors, so it is difficult to realize automation

Method used

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  • Method for detecting treponema pallidum, quantum-dot labeled immunochromatography test paper and preparation method thereof
  • Method for detecting treponema pallidum, quantum-dot labeled immunochromatography test paper and preparation method thereof

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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 treponema pallidum monoclonal antibody glass cellulose membrane B, absorbent paper, described Glass cellulose membrane A is the glass cellulose membrane purchased on the market without point sample;

[0033] Wherein, glass cellulose membrane A, glass cellulose membrane B of quantum dot-labeled Treponema pallidum monoclonal antibody, nitrocellulose membrane, and absorbent paper are pasted on the plastic plate in sequence;

[0034] Wherein, there are Treponema pallidum 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 quantum dot-labeled Treponema pallidum monoclonal antibody is located at one end of the glass cellulose membrane B, corresponding to the detecti...

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 Treponema pallidum 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 Treponema pallidum 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 Treponema pallidum polyclonal antibody and rabbit anti-mouse secondary antibody with 0.05-0.15M PBS buffer, spray 0.5g / L Treponema pallidum polyclonal antibody and 1.0g / L rabbit anti-mouse secondary antibody...

Embodiment 3

[0058] Embodiment 3: using the test paper to detect Treponema pallidum, comprising the following steps: spotting the sample on the assembled test paper near one end of Treponema pallidum, reacting for 5 minutes, and observing the result in an ultraviolet analyzer. PBS buffer solution and normal human blood were used as blank controls.

[0059] Result judgment: On 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 medical immunodetection method and particularly relates to a method for using quantum-dot labeled immunochromatography test paper to detect treponema pallidum by an immunological method. The quantum-dot labeled immunochromatography test paper is characterized in that a plastic board is stuck with a glass cellulose membrane A, a glass cellulose membrane B of a quantum-dot labeled treponema pallidum IgG monoclonal antibody, a nitrocellulose membrane and water absorbing paper from bottom to top in sequence, wherein one end of the nitrocellulose membrane is provided with treponema pallidum polyclonal antibody and rabbit antimouse second antibody so as to form a detecting band T and a quality control band C; the quantum-dot labeled treponema pallidum IgG monoclonal antibody is positioned at one end of the glass cellulose membrane and corresponds to the detecting band T and the quality control band C, and the quantum-dot labeled treponema pallidum IgG monoclonal antibody is positioned at one end of a sampling point. The method has the advantages that high specificity of immunoreactions and the fluorescence characteristic of quantum dots are combined, so that the detection sensitivity is about 1000 times higher than that of the current commonly-used colloidal gold detection method.

Description

technical field [0001] The invention relates to a medical immunological detection method, in particular to a method for detecting Treponema pallidum with an immunological method by using quantum dot-labeled immunochromatographic test paper. Background technique [0002] Treponema pallidun (Treponema Pallidun) is the pathogen of syphilis, because it is transparent and not easy to color, it is also called treponema pallidun. Syphilis is a widely prevalent venereal disease, and its incidence has risen again in China. Treponema pallidum only infects humans, divided into acquired syphilis and congenital syphilis. Acquired syphilis is mainly transmitted through sexual contact; congenital syphilis is transmitted from Treponema pallidum through the placenta to the fetus from the umbilical cord blood circulation, which can cause systemic infection of the fetus. Spirochetes reproduce in large numbers in the internal organs and tissues of the fetus, which can cause fetal death or mis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/533
CPCG01N33/571
Inventor 文德敏申有长于晓永
Owner 北京华卫骥生物医药有限公司
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