Test paper box capable of detecting escherichia coli O157:H7

A technology of O157 and Escherichia coli, applied in the field of bacteriology and immunology, can solve the problems of human health and safety impact, obvious material matrix effect, poor stability, etc., and achieve the effects of cost saving, simple process and convenient preparation.

Inactive Publication Date: 2015-01-28
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006]3. The colloidal gold labeling process is a physical adsorption process, so the stability in the liquid phase is poor;
[0007]4. Only when the gold particles gather to a certain amount, the naked eye can observe the purple-red band, and the color band has little contrast with the background, thus limiting Improve the detection sensitivity;
[0008]5. The matrix effect of different materials is obvious, and the background interference is very large
Antigen pairing process takes a long time, increasing R&D costs

Method used

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  • Test paper box capable of detecting escherichia coli O157:H7
  • Test paper box capable of detecting escherichia coli O157:H7
  • Test paper box capable of detecting escherichia coli O157:H7

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 : Single antibody test strip structure description and preparation method of the present invention

[0031] 1. The test strip box of the present invention

[0032] The test paper box for detecting Escherichia coli O157:H7 includes a single antibody test strip and a fluorescent dye box; the single antibody test strip includes a sample pad, a nitrocellulose membrane, an absorbent pad and a PVP base plate, and its specific structural features are : The nitrocellulose membrane is pasted on the PVP base plate; one end of the base plate is a sample pad, and the other end is an absorbent pad; the width is 3-4mm / cut and then a single antibody test strip; the nitrocellulose The membrane is sprayed with Escherichia coli O157:H7 antibody as the detection line T line, and the anti-fluorescent dye antibody spray quality control line C line;

[0033] Fluorescent dye cartridges contain the fluorescent dyes propidium iodide, fluorescein diacetate (FDA), acridine orange, or...

Embodiment 2

[0046] Example 2 : Test strips after bacteria stained with other dyes

[0047] Common dyes saffron, sand yellow, crystal violet and common fluorescent dye DiOC 18 (3) After staining Escherichia coli O157:H7 with fluorescein isothiocyanate (FITC) for 10 minutes, put the sample on the test strip of the present invention. Spray the corresponding dye antibody on the C line;

[0048] The result is as image 3 As shown, other dyes were found to be unable to develop color on the test strip. It is speculated that these dyes are not suitable for use in test strips. When they combine with the bacteria, they also combine with the nitrocellulose membrane or sample pad of the test strip, so that the stained bacteria cannot migrate on the test strip. , and finally cannot develop color. And four kinds of fluorescent dyes of propidium iodide, fluorescein diacetate (FDA), acridine orange or SYBR Green screened by the present invention can be preferably suitable for the use of test strips...

Embodiment 3

[0049] Example 3: Test paper box of the present invention detects Escherichia coli O157: H7

[0050] Get the test paper box that embodiment 1 prepares;

[0051] The number of cells taken was 10 2 、10 3 、10 4 、10 5 、10 6 、10 7 、10 10 Escherichia coli O157:H7 suspension was stained with 0.01% fluorescent dye solution for 5 min. Add 4 drops of the sample (about 100 μL) to the sample hole of the test strip after the stained bacterial liquid. After reacting for 15 minutes, judge the result under the ultraviolet light. There are two bands at the detection line, and the result is positive. There is no bright band in the detection line, and the result is negative;

[0052] Compared with the detection results of Escherichia coli O157:H7 colloidal gold test strips with the same number of bacteria, Figure 4 Escherichia coli O157:H7 colloidal gold test strip test results show that the lowest detection limit is 10 bacteria 5 to 10 4 between. Escherichia coli O157:H7 single ...

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Abstract

The invention relates to an immunochromatography test paper box based on a thallus fluorescence staining technology and used for detecting a stained escherichia coli O157:H7 thallus by using a single antibody. The immunochromatography test paper box comprises a single antibody test strip and a fluorescence dye box. The immunochromatography test paper box has the advantages that the escherichia coli O157:H7 thallus is pre-stained by using a fluorescence dye, and a specific antibody of the escherichia coli O157:H7 is sprayed on a test strip T line to capture the escherichia coli O157:H7 thallus in a mixed sample, so that the thallus in the sample is rapidly and specifically detected.

Description

technical field [0001] The invention relates to the technical fields of bacteriology and immunology. Specifically, the invention relates to an immunochromatographic test strip for detecting bacteria with a single antibody and a preparation method thereof. Background technique [0002] Immunochromatography (immunochromatography) is a rapid detection technology established on the basis of immunofiltration in the late 1980s and early 1990s. The antibody (or antigen) of the antigen-antibody reaction is pre-coated on the specific position of the immunochromatographic test paper in the form of a strip, and the antigen (or antibody) in the sample to be tested is pre-labeled by the pigment during the process of developing with the developing solution. When the antigen (or antibody) in the sample to be tested marked by the dye is expanded to a specific position (that is, the position where the antibody (or antigen) is coated in a strip), the antigen (or antibody) in the sample to be...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 徐锋魏华王登远武晓丽叶若松甘敏陈星星杨栋刘琚珥许恒毅胡海宁
Owner NANCHANG UNIV
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