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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 fast and sensitive detection, inconvenient operation, etc., and achieve easy interpretation of results, simple operation, and convenient production. easy effect

Active Publication Date: 2013-04-17
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
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Effect test

Embodiment 1

[0034] see Figure 1 to Figure 3 , a colloidal gold immunochromatography detection kit based on a nucleic acid aptamer, comprising a detection test paper, the detection test paper comprising a base plate 1, a sample pad 2 bonded to the base plate 1 and overlapped sequentially, a binding pad 3, Nitrocellulose membrane 4 and water-absorbing pad 7; On the described nitrocellulose membrane 4, a detection line 5 is provided near the side of the binding pad 3, and on the nitrocellulose membrane 4, a side near the water-absorbing pad 7 is provided with a quality control line 6; A nucleic acid aptamer labeled with nanoparticles is coated on the binding pad 3; a complex of nucleic acid aptamer B and streptavidin is coated on the detection line 5; streptavidin is coated on the quality control line 6 Hedin; the nucleic acid aptamer A and the nucleic acid aptamer B are biotin-labeled nucleic acid aptamers that can specifically recognize the same detection object; wherein, the nanoparticle...

preparation Embodiment 1

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

[0038] (1) Aptamer 1 labeled colloidal gold particles: Centrifuge the synthesized 13nm colloidal gold solution at 10,000rpm for 15 minutes, suck out the supernatant, add the original volume of sterile water, repeat the operation twice, and add 0.5 The ratio of the OD nucleic acid aptamer, the DNA sequence of the 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, and slowly add it to the colloidal gold solution under magnetic stirring. After standing at room temperature for 24 hours, add 0.1 M PB solution to adjust the final concentration of PB to 10mM; add PBS buffer solution to adjust the solution. The concentration of NaCl was 0.1M, aged at room temperature for 24 hours; finally, PBS buffer was added to adjust the final concentration of NaCl in the solution to 0.3M, after aging at room temperature...

Embodiment 3

[0045] see Figure 1 to Figure 3 , a colloidal gold immunochromatography detection kit based on a nucleic acid aptamer, comprising a detection test paper, the detection test paper comprising a base plate 1, a sample pad 2 bonded to the base plate 1 and overlapped sequentially, a binding pad 3, Nitrocellulose membrane 4 and water-absorbing pad 7; On the described nitrocellulose membrane 4, a detection line 5 is provided near the side of the binding pad 3, and on the nitrocellulose membrane 4, a side near the water-absorbing pad 7 is provided with a quality control line 6; A nucleic acid aptamer labeled with nanoparticles is coated on the binding pad 3; a complex of nucleic acid aptamer B and streptavidin is coated on the detection line 5; streptavidin is coated on the quality control line 6 Hedin; the nucleic acid aptamer A and the nucleic acid aptamer B are biotin-labeled nucleic acid aptamers that can specifically recognize the same detection object; wherein, the nanoparticle...

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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 present invention relates to a medical detection product and a production method thereof, in particular to a nucleic acid aptamer-based chromatography detection kit and a preparation method and detection method thereof. Background technique [0002] The common immunoassay method is to immobilize a specific labeled antibody on a solid support first, specifically bind 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 realize the specific immunodiagnosis. [0003] The biological macromolecules used in the commonly used immunochromatography method to specifically recognize the detection object are antibodies. Through the specific recognition reaction of antigen and antibody, the immune colloidal particles marked on the antigen or antibody make the detection line and quality control line display a certain color, thereby Realize specific im...

Claims

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Application Information

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IPC IPC(8): G01N33/566G01N33/531G01N33/558
Inventor 谭蔚泓常荣山蒋健晖
Owner 谭蔚泓
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