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Fluorescence detection kit based on aptamer

A fluorescence detection, nucleic acid aptamer technology, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of difficult design, limited scope of use, complexity, etc. Effect

Active Publication Date: 2018-04-03
GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
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  • Abstract
  • Description
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Problems solved by technology

[0003] Existing nucleic acid aptamer-based detection methods are generally more complicated, and it is difficult to detect different substances by changing the nucleic acid aptamer sequence, which makes its design more difficult and limits its scope of application

Method used

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  • Fluorescence detection kit based on aptamer
  • Fluorescence detection kit based on aptamer
  • Fluorescence detection kit based on aptamer

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Embodiment 1

[0055] A detection kit for malathion residues, comprising:

[0056] (1) Nucleic acid DNA1, DNA2, stem-loop nucleic acid H1, the sequences of which are as follows:

[0057] Malathion aptamer DNA1: 5'-ATCCGTCACACCTGCTCTTATACACAATTGTTTTTCTCTAACTTCTTGACTGCTGGTGTTGGCTCCCGTAT-3';

[0058] DNA2: 5'-AAGTTAAGAGAAAAAACAATTG ACACAC -3';

[0059] H1: 5'-CAATTGTTT(a)-GCTTACGGT(b)-ATACCTACGTCGATCGCA(c)-ACCGTAAGC(b*)-TTCTCTTAACTT(d)-3';

[0060] (2) Exonuclease III

[0061] (3) Malathion standard solution

[0062] (4) Reaction buffer, containing 15mM Tris-HCl, 50mM NaCl, 10mM MgCl 2 , pH 7.0.

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Abstract

The invention discloses a fluorescence detection kit based on an aptamer. The fluorescence detection kit comprises exonuclease III, a reaction buffer solution, DNA1, DNA2 and stem-loop structure nucleic acid H1. The aptamer is used for specific recognition of a molecular recognition element, a 3' double-chain nucleic acid structure with a flush end is obtained, and circular signal amplification isperformed after exonuclease III is added, so that a large amount of double-chain DNA become single-chain DNA; after fluorescent dye is added, the content of to-be-detected molecules is detected by distinguishing fluorescence intensity change. The linear range for malathion detection is 10 pM to 100 nM, and the limit of detection is 5 pM; meanwhile, the detection system has good specificity and the detection results cannot be interfered by other common analogues. No separation and purification processes are required in the detection process, the kit can be used for detection after simple mixing, has the advantages of being simple to operate, low in cost, rapid in response and the like and can be used for rapid and high-sensitivity detection of pesticide residues in food, water sources, soil and other samples.

Description

technical field [0001] The invention relates to a detection kit, in particular to a nucleic acid aptamer-based fluorescence detection kit. Background technique [0002] Nucleic acid aptamer (aptamer) is an oligonucleotide fragment that can specifically bind to proteins or other small molecular substances screened by the in vitro screening technology SELEX (Systematic Evolution of Ligands by Exponential Enrichment). As a new type of molecular recognition tool, nucleic acid aptamer not only has high affinity and specificity, but also has the advantages of easy synthesis, easy modification, and good stability compared with traditional antibodies. It has a good application in the field of protein and molecular detection. application potential. In recent years, combined with the molecular characteristics of nucleic acid aptamers, the detection of proteins has been converted into the detection of specifically recognized nucleic acid aptamers, which not only avoids the shortcoming...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/533G01N33/53
CPCG01N33/5308G01N33/533G01N33/68G01N2333/47
Inventor 陈俊华周丹华潘家峰王琦
Owner GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
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