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Brucella abortus virus fluorescent microsphere immunochromatography test paper and preparation method and detection method thereof

A technology of immunochromatographic test paper and fluorescent microspheres, which can be used in measurement devices, analytical materials, instruments, etc., can solve the problems of low sensitivity, cumbersome and time-consuming steps, single color, etc., and achieve the effect of high detection sensitivity and simple operation requirements.

Inactive Publication Date: 2019-04-19
GUANGZHOU VETERINARY BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the above methods all require advanced detection instruments, the detection costs are expensive, the steps are cumbersome and time-consuming, and the requirements for the professionalism of the operators are high, they are not suitable for a large number of rapid screening tests in grassroots enterprises and institutions.
[0004] Colloidal gold immunochromatography has the characteristics of fast detection speed, low price, and simple operation, but there are still some defects, such as poor stability, low sensitivity, quantitative detection cannot be realized, matrix effect is obvious, background interference is large, and the color is single, making it difficult to achieve multiple inspection or joint inspection

Method used

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  • Brucella abortus virus fluorescent microsphere immunochromatography test paper and preparation method and detection method thereof
  • Brucella abortus virus fluorescent microsphere immunochromatography test paper and preparation method and detection method thereof
  • Brucella abortus virus fluorescent microsphere immunochromatography test paper and preparation method and detection method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A kind of bovine brucellosis virus fluorescent microsphere immunochromatographic test paper provided by the present invention, refer to figure 1 , including a bottom plate 5, the bottom plate 5 is connected with a sample pad 1, a binding pad 2, a chromatography film 3 and a water absorption pad 4 in turn, the chromatography film is provided with a C line and a T line, the combination Pad 2 is coated with fluorescent microspheres to be labeled with BrG11 antibody; the T line is coated with BrF11 antibody; the C line is coated with goat anti-mouse IgG antibody.

Embodiment 2

[0036] The preparation method of the bovine brucellosis virus fluorescent microsphere immunochromatographic test paper in the implementation 1 is:

[0037] Materials and Methods

[0038] 1. Antibodies, detection antigens, fluorescent microsphere reagents and instruments

[0039] BrG11 and BrF11 were purchased from Guangzhou Jianyang Biotechnology Co., Ltd.;

[0040] The brucellosis vaccine was purchased from Harbin Pharmaceutical Group Biological Co., Ltd. for positive test samples;

[0041] 210nm fluorescent microspheres (Item No. MD021) were purchased from Nanjing Microtest Biotechnology Company;

[0042] Fluorescence immunoassay analyzer was purchased from Nanjing Microtest Biological Company;

[0043] The XYZ axis spray gold filming instrument was purchased from Jinbiao Biology;

[0044] 2. Antibody-conjugated labeled fluorescent microspheres

[0045] According to the principle of carboxyl-activated coupling, the antibody is coupled to labeled fluorescent microspheres...

Embodiment 3

[0049] Detection of positive samples with bovine brucellosis fluorescent microsphere immunochromatographic test paper

[0050]Dilute the bovine brucellosis antigen into solutions with concentrations of 1×106CFU / ml, 2×106CFU / ml, 4×106CFU / ml~4096×106CFU / ml, 8192×106CFU / ml, 16384×106CFU / ml, Each get 100 μ l drop-wise respectively on the sample pad of the bovine brucellosis fluorescent microsphere immunochromatography test paper prepared in embodiment 2, after reacting for 15 minutes, detect the fluorescent signal with a fluorescence detector (Shanghai Weicei Biotechnology Co., Ltd.) that emits 365nm laser , and record the T value (fluorescence signal intensity of T line), C value (fluorescence signal intensity of C line), and T / C value (ratio of fluorescence signal intensity of T line to C line) of each group.

[0051] Table 1 Result of detecting positive samples

[0052]

[0053]

[0054] 5. Generate a fitting curve of the relationship between T / C value and concentration ...

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Abstract

The invention discloses brucella abortus virus fluorescent microsphere immunochromatography test paper and a preparation method and detection method thereof. The invention aims to provide the brucellaabortus virus fluorescent microsphere immunochromatography test paper which has strong specificity, high sensitivity and good repeatability and can be used for quantitative and qualitative analysis.According to the technical scheme, the brucella abortus virus fluorescent microsphere immunochromatography test paper comprises a bottom plate, the bottom plate is sequentially connected with a samplepad, a combination pad, a chromatographic membrane and a water absorption pad, a line C and a line T are arranged on the chromatographic membrane, and the combination pad is sprayed with fluorescentmicrospheres labeled with a BrG11 antibody; a BrF11 antibody is sprayed on the line T; and the line C is painted with A goat anti-mouse IgG antibody. The invention belongs to the field of pathogen detection of experimental animals.

Description

technical field [0001] The invention belongs to the field of pathogen detection of experimental animals, in particular to a fluorescent microsphere immunochromatographic test paper of bovine brucellosis virus, and also relates to a preparation method of the detection test paper. Background technique [0002] Brucellosis is currently one of the most widespread and harmful zoonotic diseases in the world. Human and animal brucellosis exists and is prevalent in more than 170 countries and regions around the world. Human and animal brucellosis exists and is prevalent in 25 provinces (cities and districts) in my country. With the rapid development of animal husbandry, brucellosis is clearly on the rise in most endemic areas. In 2005, there were 19,664 new cases nationwide, with an incidence rate of 1.504 / 100,000, exceeding the historical record. Brucella is an intracellular gram-negative bacterium, which mainly causes wavelike fever and chronic infection in humans, as well as or...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/558G01N33/533G01N33/543
CPCG01N33/533G01N33/54313G01N33/558G01N33/56983
Inventor 关剑池伍绮文罗梦珍潘玉
Owner GUANGZHOU VETERINARY BIOTECH CO LTD
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