Preparation and detection methods of fluorescent quantitation high-sensitivity visual joint inspection immunochromatography test strip based on magnetic enrichment

An immunochromatographic test strip and fluorescence quantitative technology, which is applied in the field of medical diagnosis, can solve the problems of light color of detection strips, limit detection sensitivity, and cannot be identified with naked eyes, etc., and achieve the effect of eliminating interference and improving accuracy.

Inactive Publication Date: 2018-02-09
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently commercialized immunochromatographic test strips mostly use colloidal gold as a marker. Although the test results are visible to the naked eye, accurate quantitative detection cannot be completed, and the detection strips of samples with an antigen concentration slightly higher than the critical value are often too light to be detected. Identify with naked eyes; Fluorescence immunochromatography test strip detection technology is a new technology emerging after colloidal gold immunochromatography test strip technology to achieve quantitative detection. An important factor in sensitivity, which includes autofluorescence from the nitrocellulose membrane and the sample itself

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  • Preparation and detection methods of fluorescent quantitation high-sensitivity visual joint inspection immunochromatography test strip based on magnetic enrichment
  • Preparation and detection methods of fluorescent quantitation high-sensitivity visual joint inspection immunochromatography test strip based on magnetic enrichment
  • Preparation and detection methods of fluorescent quantitation high-sensitivity visual joint inspection immunochromatography test strip based on magnetic enrichment

Examples

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Embodiment example 1

[0039] Preparation method of fluorescent quantitative high-sensitivity visual joint detection immunochromatographic test strip detection technology based on magnetic enrichment: Spray fluorescein Cy5-labeled breast cancer-related markers CEA and CA153 coated antibodies on the detection line position of the nitrocellulose membrane , the goat anti-mouse antibody solution was sprayed on the quality control line position of the nitrocellulose membrane, the mass fraction ratio of polymer PLGA and magnetic nanoparticles was 15:1, and the mass fraction ratio of EDC and magnetic nanoparticles was 1: 60. The amount of BSA blocking solution was 2%, and 200 μL of the mixed antigen of CA153 with a concentration of 50 U / mL and CEA with a concentration of 10 ng / mL was enriched in the sample pool, and three replicates were performed. The qualitative detection method is as follows: After adding the above-mentioned sample test strips to react for 2 minutes, observe the reading frame with the na...

Embodiment example 2

[0041] Preparation method of fluorescent quantitative high-sensitivity visual joint detection immunochromatographic test strip detection technology based on magnetic enrichment: Spray fluorescein Cy5-labeled breast cancer-related markers CEA and CA153 coated antibodies on the detection line position of the nitrocellulose membrane , the goat anti-mouse antibody solution was sprayed on the quality control line position of the nitrocellulose membrane, the mass fraction ratio of polymer PLGA and magnetic nanoparticles was 10:1, and the mass fraction ratio of EDC and magnetic nanoparticles was 1: 60. The amount of BSA blocking solution was 2%, and 200 μL of the mixed antigen of CA153 with a concentration of 50 U / mL and CEA with a concentration of 10 ng / mL was enriched in the sample pool, and three replicates were performed. The qualitative detection method is as follows: After adding the above-mentioned sample test strips to react for 2 minutes, observe the reading frame with the na...

Embodiment example 3

[0043] Preparation method of fluorescent quantitative high-sensitivity visual joint detection immunochromatographic test strip detection technology based on magnetic enrichment: Spray fluorescein Cy5-labeled breast cancer-related markers CEA and CA153 coated antibodies on the detection line position of the nitrocellulose membrane , the goat anti-mouse antibody solution was sprayed on the quality control line position of the nitrocellulose membrane, the mass fraction ratio of polymer PLGA and magnetic nanoparticles was 20:1, and the mass fraction ratio of EDC and magnetic nanoparticles was 1: 60. The amount of BSA blocking solution was 2%, and 200 μL of the mixed antigen of CA153 with a concentration of 50 U / mL and CEA with a concentration of 10 ng / mL was enriched in the sample pool, and three replicates were performed. The qualitative detection method is as follows: After adding the above-mentioned sample test strips to react for 2 minutes, observe the reading frame with the na...

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Abstract

The invention relates to preparation and detection methods of a fluorescent quantitation high-sensitivity visual joint inspection immunochromatography test strip based on magnetic enrichment. Fe3O4 magnetic nanoparticles are subjected to multiple wrapping through a macromolecule polylactic acid-hydroxyacetic acid copolymer by an ultrasonic emulsification method, so that a target substance can be fast and effectively combined onto a magnetic microsphere; a newly generated compound has the same magnetic responsibility in the magnetic field, so that samples to be inspected are enriched; the detection sensitivity is enhanced; meanwhile, the joint detection is realized; the double-antibody sandwich immunoreaction is performed by an immunochromatography test strip detection platform; the reaction mechanism of fluorescence signals can be quenched through magnetic nanoparticles under the specific conditions, so that rapid qualitation under the naked eye condition and precise and quantitative wide-range bimodal detection in dark field test are realized; meanwhile, the superparamagnetism of the magnetic nanoparticles is used for enriching serum to be detected; the accuracy of breast cancer early diagnosis and the universality of community screening are improved through the joint detection of breast cancer tumor markers CEA (carcinoembryonic antigen) and CA153.

Description

technical field [0001] The invention relates to the technical field of medical diagnosis, and more specifically relates to a preparation and detection method of a fluorescent quantitative high-sensitivity visual joint detection immunochromatography test strip based on magnetic enrichment. Background technique [0002] Breast cancer is the most common and most frequent malignant tumor in modern women. It has now taken the first place in the incidence of malignant tumors in women, and the incidence rate is increasing every year. According to statistics, in most developed countries, malignant tumors are the leading cause of death among residents. First or second place in the middle. Among them, the incidence of breast cancer ranks fifth among the eight major cancers in China. With the rapid economic development, breast cancer patients are also increasing by leaps and bounds. Whether distant metastasis occurs after breast cancer surgery is the main factor determining the surviva...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N33/558G01N33/574
Inventor 常津张博宫晓群高玮辰
Owner TIANJIN UNIV
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