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Novel ligand detecting method

A detection method and a ligand technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems such as low sensitivity, poor accuracy, and complicated operation, and achieve high sensitivity, easy operation, and simple operation.

Inactive Publication Date: 2004-08-18
甘肃省医学科学研究院
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is complicated to operate, needs to separate the ligand and the ligand, and has low sensitivity (due to the purity of the ligand and the recombination of the ligand and the ligand after the separation of the ligand and the ligand to block DNA replication) and poor accuracy.

Method used

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Embodiment 1

[0037] refer to figure 1 , using the ligand-specific oligonucleotide ligand modified by the nucleic acid sequence to directly bind the ligand, the ligand signal is converted into a nucleic acid signal, and then amplified by PCR to detect the ligand.

[0038] The principle and technical route of this embodiment are: fix the ligand to be tested on a medium such as a nitrocellulose membrane, seal the nitrocellulose membrane with a blocking solution, and then place the membrane with the ligand-specific oligonucleotide modified by nucleic acid sequence The acid ligand is incubated for a certain period of time, the cellulose membrane is fully washed with washing solution, the unbound specific oligonucleotide ligand is removed, and the ligand-specific oligonucleotide modified by the nucleic acid sequence combined with the ligand is amplified by PCR technology Acid ligand. The primers used in PCR amplification are labeled with fluorescent reagents or isotopes, etc., and the detection...

Embodiment 2

[0041] No need for any linking molecules, but the ligand-specific oligonucleotide ligands modified by chemiluminescent substances, enzymes, fluorescence and isotopes are directly combined with the ligands, and the ligand signals are converted into molecular signals, and then through the molecular signal Recognition, detection of ligands.

[0042] The principle and technical route of this embodiment are: fix the ligand to be tested on a medium such as a nitrocellulose membrane, seal the nitrocellulose membrane with a blocking solution, and then combine the membrane with a ligand-specific oligonucleotide modified by a signal molecule The acid ligand is incubated for a certain period of time, and the cellulose membrane is fully washed with washing solution to remove the unbound specific oligonucleotide ligand, and the detection of the ligand signal is realized by detecting the intensity of fluorescence or isotope. The present invention emphasizes that in the ligand detection meth...

Embodiment 3

[0044] After the oligonucleotide ligand of a certain type of ligand is artificially modified by the nucleic acid sequence (such as artificially adding some nucleic acid sequences that do not affect its binding activity), according to the length of the modified sequence, the base sequence is different, through the oligonucleotide The specific combination of the nucleotide ligand and the ligand makes the modified sequence represent different ligands, and then amplified by PCR (or rolling circle replication) to detect the modified sequence, thereby identifying the ligand. This technology can be used to build microarrays or biochips.

[0045]The principle and technical route of this embodiment are: fix the test samples containing various ligands to be tested on media such as nitrocellulose membranes, seal the nitrocellulose membranes with blocking solution, and then combine the membranes with those modified with different nucleic acid sequences. After incubating the specific oligo...

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Abstract

The present invention relates to one new kind o detecting ligand of nucleic acid aglucone. The method utilizes nucleic acid sequence modified ligand to specify oligonucleotide aglucone for direct binding with ligand and converting ligand signal into nucleic acid aglucone signal. The ligand is then determined via subsequent PCR proliferation and amplification and detection. The method of detecting ligand has the features of fast speed, high sensitivity, powerful specificity multi-ligand microarray detection.

Description

technical field [0001] The invention relates to a novel ligand detection method utilizing nucleic acid ligands, in particular to a ligand detection method established by a ligand-specific oligonucleotide ligand modified by a nucleic acid sequence. 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 purpose. In recent years, alth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 廖世奇邵宁生韩庆斌张雪力王黎王晓辉张春梅吴志豪
Owner 甘肃省医学科学研究院
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