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Temperature type biosensor and method for detecting target aptamer by using temperature type biosensor

A biosensor and aptamer technology, applied in biochemical equipment and methods, microbial measurement/inspection, biomaterial analysis, etc., to achieve the effect of wide detection range, high selectivity temperature detection, and low lower limit

Active Publication Date: 2021-08-17
JIANGXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In recent years, the traditional methods for detecting malathion mainly include electrochemical methods, fluorescence analysis, photoelectrochemical analysis, liquid chromatography, etc. ([1] S. Uniyal, R. Sharma. Technological advancement inelectrochemical biosensor based detection of organophosphate pesticidechlorpyrifos in the environment: a review of status and prospects[J].Biosensors and Bioelectronics,2018,116:37-50.[2]H.Li,X.Yan,G.Lu,et al.Carbondot-based bioplatform for dual colorimetric and fluorometric sensing of organophosphate pesticides[J].Sensors and Actuators B:Chemical,2018,260:563-570.[3]J.Tang,J.Li,P.Xiong,et al.Rolling circle amplification promoted magneto-controlled photoelectrochemical biosensor for organophosphorus pesticides based on dissolution of core-shell MnO 2 nanoflower@CdS mediated bybutyrylcholinesterase[J].Microchimica Acta,2020,187(8):450.[4]E.Cequier,A.Sakhi,L.Haug,et al.Development of an ion-pair liquid chromatography-high resolution mass spectrometry method for determination of organophosphatepesticide metabolites in large-scale biomonitoring studies[J].Journal of Chromatography A,2016,1454:32-41.) However, due to complex instruments and the needs of professionals, the practical application of most detection methods is still restricted

Method used

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  • Temperature type biosensor and method for detecting target aptamer by using temperature type biosensor
  • Temperature type biosensor and method for detecting target aptamer by using temperature type biosensor
  • Temperature type biosensor and method for detecting target aptamer by using temperature type biosensor

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Embodiment 1

[0043] (1) Disperse 100 μM S-A and S-B with acrylamide groups in 10 mM Tris-HCl buffer (pH=7.4) containing 4% acrylamide, 200 mM NaCl, 1 mM EDTA, respectively, and bubble with nitrogen for 5 min to remove the air. Then, 1.4% (v / v) freshly prepared catalyst (0.05 g APS, 25 μL TEMED, 0.5 mL deionized water) was added to the above mixture to promote the polymerization of acrylamide to form polyacrylamide polymer. Then, nitrogen bubbling was carried out for 5 min to form polymer chain A (PS-A) and polymer chain B (PS-B); after that, CAT (300 U) was added thereto, and PS was mixed with 1:1:1 molar ratio - A and PS-B were mixed with malathion aptamer (both at a concentration of 0.1 mM), and nitrogen gas was bubbled for 5 min. Finally, the mixed solution was incubated at 25 °C for 20 min to form a 3D DNA hydrogel cross-linked by aptamers that encapsulated CAT.

[0044] (2) Incubate 10 μL of different concentrations of target malathion with the 3D DNA hydrogel prepared in step (1) f...

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Abstract

The invention discloses a temperature type biosensor and a method for detecting a target aptamer by using the temperature type biosensor. The temperature type biosensor comprises an oxygen production bottle and a heat production bottle which are communicated through a conduit, aptamer response type DNA hydrogel is arranged at the bottom of the oxygen production bottle, and a liquid feeder is arranged at the top of the oxygen production bottle; an oxidized heating material is arranged at the bottom of the heat production bottle, and a thermometer is inserted into the top of the heat production bottle; the aptamer response type DNA hydrogel is formed by wrapping catalase with aptamer cross-linked type DNA hydrogel. The temperature type biosensor utilizes specific recognition between an aptamer and a target aptamer, adopts a DNA hydrogel amplification mechanism and utilizes aptamer response type DNA hydrogel to release CAT under stimulation of the target aptamer so as to catalyze H2O2 to generate O2, O2 and an oxidation heating material are subjected to an oxidation reduction reaction to release a large amount of heat, so that temperature change is caused, therefore, the temperature type biosensor is constructed to achieve high-sensitivity and high-selectivity temperature detection of the target aptamer.

Description

technical field [0001] The invention relates to a temperature-type biosensor, in particular to a temperature-type biosensor based on an aptamer-responsive DNA hydrogel and an oxidative heating material, and also to a method for detecting a target aptamer using a temperature-type biosensor, which belongs to the traditional Sensitive analysis technology field. Background technique [0002] To protect crops from pests and insects, pesticides play a vital role in modern agriculture and aquaculture. Most agricultural production worldwide relies on the use of organophosphorus pesticides (OPs) to increase crop productivity. Compared with other pesticides, malathion is widely used in agriculture because of its high efficiency and low toxicity, so as to reduce the loss of agricultural production. However, if malathion is used excessively, it may cause serious problems to food safety, the environment, and human health. According to reports, in 2013, a high concentration of malathio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N25/22G01N31/10C12Q1/30
CPCG01N25/22G01N31/10C12Q1/30G01N2333/908
Inventor 汤娟程宏丽覃娇刘丽萍高珊
Owner JIANGXI NORMAL UNIV
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