D-dimer quantitative fluorescence immunoassay test strip and preparation method thereof

A fluorescence immunoassay and quantitative assay technology, applied in the field of medical immunology in vitro diagnosis, can solve the problems of not providing data, not knowing whether the kit is real and feasible, and inaccurate assay results, and achieving the effects of simple operation, low cost and high sensitivity

Inactive Publication Date: 2012-09-26
GETEIN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, measurement results are easily affected by manipulation and are inaccurate
In addition, in this invention, only the results are provided with r

Method used

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  • D-dimer quantitative fluorescence immunoassay test strip and preparation method thereof
  • D-dimer quantitative fluorescence immunoassay test strip and preparation method thereof
  • D-dimer quantitative fluorescence immunoassay test strip and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The preparation method of the D-dimer fluorescent immunoassay test strip is as follows:

[0041] 1) Preparation of fluorescent latex microspheres

[0042] Preparation of fluorescent latex microspheres: Dilute latex microspheres with a particle size of 100 nm to a final concentration of 20 mg / ml with an adsorption buffer (50 mM, citrate buffer at pH 5.8) to obtain a latex microsphere suspension ; Add appropriate amount of fluorescein Cy5-labeled streptavidin (purchased from Shanghai Yanjing Biotechnology Co., Ltd.) in the adsorption buffer, the final volume is 5ml; In the adsorption buffer of mycoavidin, the mixed solution was prepared; the resulting mixed solution was incubated at room temperature for 1-2 hours, and stirred continuously, and then centrifuged to collect the precipitate, and the precipitate was stored in buffer solution (containing 0.05% BSA for adsorption Buffer) was dissolved and stored at 4°C for later use.

[0043] 2) Preparation of biotinylated ant...

Embodiment 2

[0058] Another D-dimer fluorescent immunoassay test strip preparation method is as follows:

[0059] 1) Preparation of fluorescent latex microspheres

[0060] Preparation of fluorescent latex microspheres: Dilute latex microspheres with a particle size of 400 nm to a final concentration of 30 mg / ml with an adsorption buffer (50 mM, citrate buffer at pH 5.8) to obtain a latex microsphere suspension ; Add an appropriate amount of red fluorescein rhodamine-labeled streptavidin (purchased from Shanghai Enzyme Biotechnology Co., Ltd.) in the adsorption buffer, the final volume is 6ml; add the above-mentioned latex microsphere suspension to the above-mentioned In the adsorption buffer of sulhodamine-labeled streptavidin, a mixed solution was prepared; the resulting mixed solution was incubated at room temperature for 1-2 hours, and kept stirring, then centrifuged, and the precipitate was collected, and the precipitate was stored with a buffer solution (containing 0.06% BSA in adsor...

Embodiment 3

[0075] Example 3 D-dimer fluorescence immunoquantitative assay test strip quantitative detection

[0076] 3.1 Draw a standard curve

[0077] Add different concentrations of D-dimer antigen standard substances (take seven different concentrations, respectively 0, 0.1, 0.5, 1, 2, 5, 10mg / L, each concentration set 3 repetitions), after 10 minutes, read the signal of detection line 7 and quality control line 8 through the fluorescent immunoquantitative analyzer Getein1100 of Nanjing Jidan Biotechnology Co., Ltd., the experimental results And analysis see Table 1:

[0078] Table 1 D-dimer standard detection results

[0079]

[0080] Draw a standard curve with the D-dimer antigen standard concentration and the measured signal average value, such as figure 2 shown.

[0081] 3.2 Detection of linear range

[0082] The test strip prepared in Example 1 of the present invention was used to perform a detection linear range experiment. Take the D-dimer standard product, dilute it...

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Abstract

The invention relates to a D-dimer quantitative fluorescence immunoassay test strip. The test strip comprises a sample pad, a bind pad, a nitrocellulose film and absorbent paper, wherein the sample pad is a two-layer sample pad; the bind pad is coated with a fluorescence latex microsphere marked anti D-dimer antibody A; the nitrocellulose film is coated with an anti D-dimer antibody B serving as an assay line and a rabbit antimouse IgG antibody serving as a control line; and the fluorescence late microsphere is prepared from latex microsphere adsorptive fluorescence marked streptavidin. The test strip can be used for accurately and quantitatively assaying the content of D-dimer in human blood plasma, and has the characteristics of convenience in operation, high accuracy, high sensitivity, low cost and the like.

Description

technical field [0001] The invention belongs to the field of in vitro medical immunodiagnosis, and in particular relates to a D-dimer fluorescent immunoassay quantitative test strip and a preparation method thereof. Background technique [0002] D-dimer is a polymer formed by coagulation of fibrinogen in blood by the action of thrombin, etc., and is a specific degradation product of cross-linked fibrin. Under physiological conditions, the normal D-dimer level of the human body is generally below 200 μg / L, and the body maintains a dynamic balance between coagulation and fibrinolysis to ensure the timely formation and removal of fibrin. If this balance is disrupted, intravascular coagulation tendency will be enhanced, fibrin aggregation, fibrin degradation products will increase, and D-dimer content will increase. Therefore, the increase of D-dimer level reflects the dual activation of coagulation and fibrinolysis system in vivo, which can be used as one of the molecular mark...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558
Inventor 苏恩本黄力王勇陈伟涂策周超
Owner GETEIN BIOTECH
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