A method for qualitative and semi-quantitative detection of antigen based on paper chip immunoreaction distance
A semi-quantitative detection and immune reaction technology, applied in the direction of measuring devices, biological testing, material inspection products, etc., can solve the problems of low sensitivity, unfavorable low-content sample determination, difficulty in meeting qualitative and quantitative requirements, etc., and achieve low cost , the effect of overcoming the limitations of color recognition
Inactive Publication Date: 2018-03-06
SHAANXI NORMAL UNIV
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Problems solved by technology
Among these analysis methods, colorimetry is the most common method. The content of the sample is determined by the change of the color on the paper chip, but this method requires the eyes to make sensitive discrimination of the color, and the resolution of the human eye does not exceed 4K, the detection sensitivity is low, it is not conducive to the determination of low-content samples, it is difficult to meet the qualitative and quantitative requirements, and specific image processing software is required to achieve the purpose of sample detection
Fluorometric assays are highly sensitive, but high background signal is difficult to overcome in paper chips
Electrodes are required in the electrochemical detection method, which is costly and complicated to operate
Method used
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Embodiment 1
[0027] 1. Use CorelDraw X6 drawing software to design the paper chip model, such as figure 1 As shown, the paper chip model has 8 circular detection areas with a diameter of 6 mm, a sample injection area with a diameter of 6 mm, and a waste liquid area. Each area is connected by a flow channel with a width of 2 mm and a length of 3mm; then use the cutting knife of Ritu CE5000-40-CRP cutting plotter to cut out paper chips on Whatman No. 1 filter paper according to the above paper chip model.
[0028] 2. Put the paper chips in a PDC-32G oxygen plasma cleaner for 4 minutes. The frequency of the oxygen plasma is 13.56MHz and the power is 100W. After the treatment, aldehyde groups for immobilizing antibodies are formed on the surface of the paper chips.
[0029] 3. Add 2.5 μL 20 μg / mL carcinoembryonic antigen capture antibody solution (prepared by adding capture antibody to PBS buffer with pH = 7.4) dropwise to the detection area of the paper chip, incubate at room temperature fo...
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Abstract
The invention discloses a method for detecting antigens in a qualitative and semi-quantitative mode based on the paper chip immunoreaction distance. The method comprises the steps that after a competitive immunoreaction is completed, residual antigens marked with horse radish peroxidase react with TMB on Whatman filter paper, different migration distances are caused due to different capillary forces, and a method for detecting antigens in the qualitative and semi-quantitative mode based on the paper chip immunoreaction distance is built. The distance generated by analyzed substances of different concentrations is measured just through a ruler, the characteristics of the substances can be judged, and the method is easy to implement, low in cost and environmentally friendly and does not need expensive detection instruments or complex steps or professional operators, and rapid, simple and convenient analysis of carcino-embryonic antigens and the like is achieved.
Description
technical field [0001] The invention belongs to the technical field of cancer marker detection, and specifically relates to a method of reacting horseradish peroxidase (HRP) with precipitated 3,3',5,5'-tetramethylbenzidine (TMB) on a paper chip The migration distance is different, and the antigen is detected qualitatively and semiquantitatively by immunoassay. Background technique [0002] Paper microfluidic chips (μPADs) were first proposed by Whitesides. Due to their low cost, simple operation, and good chemical compatibility, they provide a broad platform for the development of paper chips. Paper chip is a micro-analysis platform that integrates immobilization, reaction, separation, and detection. It is convenient and fast. It has been widely used in the detection of disease markers, such as prostatic protein, immunoglobulin, carcinoembryonic antigen, and alpha-fetoprotein. For the paper chip itself, it is a common fiber material, the surface contains a large number of h...
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IPC IPC(8): G01N33/68G01N33/574G01N33/558G01N33/535
CPCG01N33/535G01N33/558G01N33/57473G01N33/57484G01N33/68G01N2333/47
Inventor 刘伟陈莹褚伟茹郭小艳
Owner SHAANXI NORMAL UNIV