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Antigen detection method and detection device made up by using said method

An antigen detection and antigen technology, applied in the field of novel antigen detection, can solve the problems of inability to perform multi-molecular detection at the same time, the operation is difficult, time-consuming, etc., and achieves considerable economic and social benefits, and improves sensitivity and high sensitivity.

Inactive Publication Date: 2003-07-09
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA +1
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Problems solved by technology

However, immuno-PCR technology has the following problems: due to the presence of connexin, a strong background signal is generated during detection (because the protein A group can bind to the Fc fragment of any antibody IgG in the sample); connexin is not commercialized; Cannot perform multiple molecular assays simultaneously
Both of the above two improved immuno-PCR techniques must use cross-linking molecules, and the Fc fragment of the antibody and the oligonucleotide binding site need to be activated before cross-linking, so the operation is difficult during the detection process and the activity of the antibody is easily lost; operation It can only be carried out under the condition of high-purity antibodies, and each antibody must be connected to DNA molecules, which is time-consuming, expensive, and difficult to popularize and apply.

Method used

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  • Antigen detection method and detection device made up by using said method
  • Antigen detection method and detection device made up by using said method

Examples

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Embodiment 1

[0027] Example 1. Immunological detection of a single antigen

[0028]Mouse IgG Fc fragment-specific oligonucleotide ligands are used to prepare mouse IgG monoclonal antibody nucleic acid labeling reagents, which are used as sensors to detect antigens recognized by the antibodies. The specific method is as follows:

[0029] Fix the antigen on the nitrocellulose membrane, and after blocking with the blocking solution, incubate the membrane with the mouse IgG monoclonal antibody and the mouse IgG Fc fragment-specific oligonucleotide ligand at 37 μg for 30 minutes, and wash with the washing solution After filtering the membrane several times, put the filter membrane back into the thin-walled PCR tube containing the PCR reaction solution (including primers and standard PCR reaction system) for PCR amplification. Fluorescent signals are detected by using fluorescent reagents labeled on the primers, and corresponding controls without PCR amplification (all corresponding reagents ...

Embodiment 2

[0030] Example 2. Preparation and application of a novel immune microarray for early diagnosis of primary liver cancer

[0031] Specific steps are as follows:

[0032] 1. Preparation of specific nucleic acid-labeled antibodies for AFP (alpha-fetoprotein), Ft (ferritin), GGT (γ-glutamyl transpeptidase) and GGT isozymes: since the above four antibodies currently used in clinical practice are all mouse IgG monoclonal antibody, so the nucleic acid sequence of the universal oligonucleotide ligand of the Fc fragment of the mouse IgG antibody is artificially modified, such as artificially adding four nucleic acid sequences A, B, C, D, the length is 40-80bp, namely become 4 kinds of specific marker nucleic acids. The four different labeled nucleic acids are combined with the above four antibodies in one-to-one correspondence, and fixed by ultraviolet cross-linking to become specific nucleic acid-labeled antibodies against the four different antigens.

[0033] 2. Prepare microarray...

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Abstract

The present invention relates to a new type antigen detection method. Said method utilizes the direct combination of antibody Fc fragment specific oligonucleotide ligand and antibody, and converts the antibody signal into nucleic acid signal, then utilizes PCR amplification technique to detect antigen. The utilization of said method can form the detection kit or develop practical protein ship or other biological chip for detecting other various antigens.

Description

technical field [0001] The invention relates to a novel antigen detection method, in particular to an antigen detection method established by using antibody Fc fragment-specific oligonucleotide ligands. The invention also relates to a detection device made by the method. Background technique [0002] Antigen detection is achieved through the effective recognition of specific determinants on antigen molecules by antibodies. Antigen detection, especially high-sensitivity detection of proteins, plays a very important role in medical research, completion of post-genome projects, and clinical testing. At present, the commonly used technology for specific antigen recognition is ELISA (enzyme-linked immunosorbent assay), which recognizes the epitope through specific antibodies, and completes the recognition and amplification of antigen information through enzymes, fluorescent substances, and radioactive isotopes connected to the antibodies. , so as to achieve the detection purpos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53
Inventor 邵宁生廖世奇柳川薛沿宁沈倍奋杨光何晓东刘元强仇杰董信春
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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