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Chromatographic detection kit based on aptamer, as well as preparation method and detection method thereof

A nucleic acid aptamer and detection kit technology, applied in measurement devices, analytical materials, instruments, etc., can solve the problems of insufficient detection speed and sensitivity, and inconvenient operation, and achieve easy interpretation of results, simple operation, and convenient production. easy effect

Active Publication Date: 2015-06-24
谭蔚泓
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method is not fast and sensitive enough, and antibody detection requires corresponding equipment, which is not easy to operate

Method used

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  • Chromatographic detection kit based on aptamer, as well as preparation method and detection method thereof
  • Chromatographic detection kit based on aptamer, as well as preparation method and detection method thereof
  • Chromatographic detection kit based on aptamer, as well as preparation method and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] See Figure 1 to Figure 3 , A nucleic acid aptamer-based colloidal gold immunochromatographic detection kit, comprising a test paper, the test paper comprising a bottom plate 1, a sample pad 2, a binding pad 3, bonded to the bottom plate 1 and sequentially overlapped The nitrocellulose membrane 4 and the absorbent pad 7; the nitrocellulose membrane 4 is provided with a detection line 5 on the side close to the bonding pad 3, and the nitrocellulose membrane 4 is provided with a quality control line 6 on the side close to the absorbent pad 7; The binding pad 3 is coated with nanoparticle labeled A nucleic acid aptamer; the detection line 5 is coated with a complex of B nucleic acid aptamer and streptavidin; the quality control line 6 is coated with streptavidin Andin; the A nucleic acid aptamer and the B nucleic acid aptamer are both biotin-labeled nucleic acid aptamers that can specifically recognize the same detection substance; wherein the nanoparticles are colloidal go...

preparation Embodiment 1

[0037] The method of preparing the kit described in Example 1:

[0038] (1) Nucleic acid aptamer 1 labeled colloidal gold particles: Centrifuge the synthesized 13 nanometer colloidal gold solution at 10000 rpm for 15 minutes, aspirate the supernatant, add the original volume of sterile water, repeat the operation twice, add 0.5 per ml of colloidal gold The ratio of OD nucleic acid aptamers, the DNA sequence of A nucleic acid aptamer is: 5'-SHC6-ATC AGG GCT AAA GAG TGC AGA GTT ACT TAG TTT TTT TTT T-3'Biotin (SEQ ID NO.1), the The aptamer can specifically recognize lysozyme. It is slowly added to the colloidal gold solution under magnetic stirring. After 24 hours at room temperature, 0.1 M PB solution is added to adjust the final concentration of PB to 10mM; then PBS buffer is added to adjust the solution The concentration of NaCl is 0.1M, aging for 24 hours at room temperature; finally add PBS buffer, adjust the final concentration of NaCl in the solution to 0.3M, and centrifuge a...

Embodiment 3

[0045] See Figure 1 to Figure 3 , A nucleic acid aptamer-based colloidal gold immunochromatographic detection kit, comprising a test paper, the test paper comprising a bottom plate 1, a sample pad 2, a binding pad 3, bonded to the bottom plate 1 and sequentially overlapped The nitrocellulose membrane 4 and the absorbent pad 7; the nitrocellulose membrane 4 is provided with a detection line 5 on the side close to the bonding pad 3, and the nitrocellulose membrane 4 is provided with a quality control line 6 on the side close to the absorbent pad 7; The binding pad 3 is coated with nanoparticle labeled A nucleic acid aptamer; the detection line 5 is coated with a complex of B nucleic acid aptamer and streptavidin; the quality control line 6 is coated with streptavidin Andin; the A nucleic acid aptamer and the B nucleic acid aptamer are both biotin-labeled nucleic acid aptamers that can specifically recognize the same detection substance; wherein the nanoparticles are colloidal go...

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Abstract

The invention discloses a chromatographic detection kit based on an aptamer, as well as a preparation method and detection method of the kit. The test paper of the kit includes a bottom board, and a sample pad, a bonding pad, a cellulose nitrate film and a water absorption pad, which are adhered on the bottom board and overlap one another; a detection line is arranged on the side of the cellulose nitrate film close to the bonding pad, and a quality control line is arranged on the side of the cellulose nitrate film close to the water absorption pad; an A aptamer labeled by colloidal gold is applied on the bonding pad; a composite containing a B aptamer and streptavidin is applied on the detection line; streptavidin is applied on the quality control line; and both of the A aptamer and the B aptamer are aptamers labeled by biotin and capable of specifically identifying a same test object. The chromatographic detection kit can display the result in five minutes only by directly dropping a diluted sample to be tested into a sample hole, is simple, quick, sensitive, easy to read result, and convenient and simple to prepare, and so on, and has the advantages of simplicity in operation and high specificity without using antibodies and instruments.

Description

Technical field [0001] The invention relates to a medical detection product and a manufacturing method thereof, in particular to a chromatographic detection kit based on a nucleic acid aptamer, and a preparation method and a detection method thereof. Background technique [0002] The common immunoassay method is to fix a specific labeled antibody on a solid support first, bind specifically to the analyte, and then bind to another antibody, so that the analyte can be qualitatively detected by the naked eye or a specific detection instrument , So as to achieve specific immunodiagnosis. [0003] Commonly used immunochromatographic methods are used to specifically identify the biological macromolecules of the test substance are antibodies, through the antigen-antibody specific recognition reaction, the immunocolloid particles labeled on the antigen or antibody make the detection line and the quality control line show a certain color, thereby Achieve specific immunodiagnosis. However,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/566G01N33/531G01N33/558
Inventor 谭蔚泓常荣山蒋健晖
Owner 谭蔚泓