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Detection assembly for detecting bacteria

A detection line, a single technology, applied in the field of detection, can solve the problems of human health and safety impact, obvious matrix effect, large background interference, etc., and achieve the effects of good specificity, simple process and high sensitivity

Inactive Publication Date: 2015-01-21
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are following deficiencies in the immunocolloidal gold chromatography test strip widely used at present: 1, colloidal gold is a kind of nanoparticle, and it is difficult to prepare colloidal gold nanoparticle, and particle diameter is difficult to control evenly
2. A strong reducing agent is used in the process of colloidal gold labeling, and at the same time, it must be exposed to heavy metal ions, which will have a certain impact on human health and safety
3. The colloidal gold labeling process is a physical adsorption process, so the stability in the liquid phase is poor
4. Only when the gold particles gather to a certain amount, the human naked eye can observe the purple-red band, and the color band has little contrast with the background, which limits the detection sensitivity
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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  • Detection assembly for detecting bacteria
  • Detection assembly for detecting bacteria
  • Detection assembly for detecting bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 The structure description and preparation method of single antibody multiple test strips of the present invention

[0048] 1. The detection combination for detecting bacteria in the present invention:

[0049] Including single antibody multiple test strips, fluorescent dye box; said single antibody multiple test strips, including sample pads, nitrocellulose membranes, absorbent pads and PVP bottom plate, its specific structural features are: nitrocellulose membranes are pasted on the PVP bottom plate One end of the bottom plate is a sample pad, and the other end is a water-absorbing pad; after being cut into strips with a width of 3-4 mm, it is a single antibody multiple test strip; the nitrocellulose membrane is sprayed with Escherichia coli O157 : H7 antibody, Listeria monocytogenes antibody, and Salmonella typhimurium antibody are used as three detection lines T1, T2, and T3 respectively, and anti-fluorescent dye antibody spray quality control line C;

[0...

Embodiment 2

[0062] Embodiment 2: the detection of the test strip after other dyes stain the thalli

[0063] Common dyes saffron, sand yellow, crystal violet and common fluorescent dye DiOC 18 (3) Three kinds of bacteria (Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium) were stained with fluorescein isothiocyanate (FITC) for 10 minutes respectively, and then loaded onto the test paper of the present invention on the strip. Spray the corresponding anti-dye antibody on the C line;

[0064] The result is as image 3 , Figure 4 , Figure 5 , Figure 6 , Figure 7 As shown, it was found that none of them could develop color on the test strip. The reason may be that these dyes are not suitable for use in test strips. When they combine with 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 dy...

Embodiment 3

[0065] Example 3: The detection combination of the present invention detects bacteria to detect Escherichia coli O157:H7

[0066] Escherichia coli O157:H7 antibody, Listeria monocytogenes antibody, and Salmonella typhimurium antibody were sprayed on the nitrocellulose membrane as three detection lines (T 1 , T 2 , T 3 Line) Fluorescent dye antibody spray quality control line (C line) spray point instrument (XYZ3000 film spotting instrument, American BIO-DOT company) (XYZ3000 film spotting instrument, American BIO-DOT company) The Y value is set as follows: 14.5, 10.5, 7.5 , 4.5cm, vacuum-dried at 37°C; paste the nitrocellulose membrane on the PVP bottom plate; one end of the bottom plate is a sample pad, and the other end is a water-absorbing pad; after cutting into strips with a width of 3-4mm, it is a single antibody multiplex test strips;

[0067] The numbers of Escherichia coli O157:H7 cells were: 1. Blank control, 2.10 4 , 3.10 5 , 4.10 6

[0068] (CFU / mL) The susp...

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Abstract

The invention relates to a detection assembly for detecting bacteria based on a bacteria fluorescent staining technique. A single antibody is utilized for detecting thalli which are stained by fluorescence; an escherichia coli O157:H7 antibody, a listeria monocytogenes antibody and a salmonella typhimurium antibody are sprayed on a test strip, and the test strip acts an immunochromatography test strip with three detection lines (T1, T2 and T3 lines). The detection assembly for detecting bacteria has the advantages that fluorochrome is used for pre-staining thalli, and the respective specific antibody is utilized again and is sprayed onto the T lines of the test strip so as to capture target thalli in a mixed sample, so that the thalli in the sample are detected rapidly and specifically; the detection assembly for detecting bacteria can be used for simultaneously detecting the food-borne pathogenic bacteria including the escherichia coli O157:H7 antibody, the listeria monocytogenes antibody and the salmonella typhimurium.

Description

technical field [0001] The invention relates to the technical field of detection. Background technique [0002] Immunochromatography technology is a rapid detection technology established on the basis of immunofiltration in the late 1980s and early 1990s. Its technical characteristics are: it will be able to react with the antigen (or antibody) in the sample to be tested. The antibody (or antigen) is pre-coated on the specific position of the immunochromatographic test strip in the form of a strip, and the antigen (or antibody) in the sample to be tested is pre-labeled by the pigment in the process of developing with the developing solution. When the antigen (or antibody) in the sample to be tested 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 tested marked by the pigment passes through the antigen-antibody The reaction is captured, so that a colored line develop...

Claims

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Application Information

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IPC IPC(8): G01N33/569
CPCG01N33/56916
Inventor 徐锋魏华武晓丽杨栋陈星星甘敏王登远陈飞刘琚珥许恒毅
Owner NANCHANG UNIV
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