PSA (Prostate Specific Antigen) high-sensitivity detection method based on nanotechnology

A detection method and nanotechnology technology are applied in the detection field of immunoenzyme-linked adsorption technology to achieve the effects of stable performance, low cost and easy analysis

Inactive Publication Date: 2012-06-27
CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0012] The technical problem to be solved in the present invention is to provide a technology that greatly improves the sensitivity of ELISA detection for the problem that the sensitivity of the current traditional ELISA detection method needs to be further improved.

Method used

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  • PSA (Prostate Specific Antigen) high-sensitivity detection method based on nanotechnology
  • PSA (Prostate Specific Antigen) high-sensitivity detection method based on nanotechnology
  • PSA (Prostate Specific Antigen) high-sensitivity detection method based on nanotechnology

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Experimental program
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Embodiment 1

[0031] The process of detection of PSA antigen by immunoenzyme-linked adsorption gold nanoparticle double-labeled probe-PCR amplification detection method

[0032] 1. Self-made 13nm colloidal gold solution.

[0033] 2. Preparation of magnetic bead probe: take 2×10 8 Each magnetic bead was operated according to the buffer solution mentioned in the manual. After washing with PBS, 6 μg of PSA polyclonal antibody was added to it, and incubated at 37°C for 4 hours, and ensured to be well mixed. Finally, it is sealed with blocking solution. Magnetic bead probes were stored at 4°C for later use.

[0034] 3. Magnetic bead probe captures PSA antigen: Add different concentrations of PSA solutions to the magnetic bead probe, the PSA concentrations are 62.5ag / ml, 1.25fg / ml, 25fg / ml, 0.5pg / ml, 10pg / ml , 200pg / ml, 4°C, protected from light, incubate overnight.

[0035] 4. Preparation of colloidal gold probe: add PSA antibody to colloidal gold, add nucleic acid after incubating at room t...

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Abstract

A PSA (Prostate Specific Antigen) high-sensitivity detection method based on nanotechnology is a novel biological high-sensitivity detection method which combines the magnetic bead probe technology, Au-nanoparticle double-mark probe technology, enzyme-linked immunosorbent assay technology and the nucleic acid amplification technology together. According to the detection method, an antibody corresponding to a protein to be detected (such as PSA) is cross-linked with a magnetic bead, and after the protein to be detected in a sample is captured, an Au-nanoparticle dual-mark probe marked by another antibody and nucleic acid is added to the captured protein. After magnetic separation and purification are carried out, nucleic acid undergoes release amplification, a visual result is obtained by means of agarose gel electrophoresis, and the grey level value scanning is subsequently performed. Compared with the conventional ELISA (Enzyme-Linked Immunosorbent Assay), the detection sensitivity of the detection method disclosed by the invention is increased by six orders of magnitude, and the PSA high-sensitivity detection method based on nanotechnology is fairly significant for the early diagnosis of caners and the detection of trace important substances, nucleic acids and proteins.

Description

technical field [0001] The present invention relates to a detection method of immunoenzyme-linked adsorption technology, in particular to a gold nanoparticle double-labeled probe-PCR (Polymerase Chain Reaction, polymerase chain reaction) that greatly improves the sensitivity of the detection method of immunoenzyme-linked adsorption technology. formula reaction) amplification detection method. Background technique [0002] Immune response is an immunological phenomenon in which members of the immune system (antigens, immune molecules, immune cells, and immune tissues) depend on each other, influence each other and interact with each other. The immunoassay method established based on the specific reaction of antigen and antibody is a method that utilizes the highly specific interaction between antigen and antibody (ie, immune response) to realize the detection of antigens, antibodies, hormones, receptors and receptors in blood or urine samples. An analytical method for the id...

Claims

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Application Information

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IPC IPC(8): G01N33/68C12Q1/68
Inventor 张玉祥孙莉
Owner CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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