CN113267529B - Temperature type biosensor and method for detecting target aptamer by using temperature type biosensor - Google Patents
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Abstract
The invention discloses a temperature type biosensor and a method for detecting a target object by using the same. The temperature type biosensor comprises an oxygen generating bottle and a heat generating bottle which are communicated through a conduit; the bottom of the oxygen generating bottle is provided with aptamer response type DNA hydrogel, and the top of the oxygen generating bottle is provided with a liquid feeder; the bottom of the heat-generating bottle is provided with an oxidation heating material, and the top of the heat-generating bottle is inserted with a thermometer; the aptamer-responsive DNA hydrogel is formed by wrapping catalase in the aptamer-crosslinked DNA hydrogel. The temperature type biosensor catalyzes H by utilizing specific recognition between an aptamer and a target aptamer and adopting a DNA hydrogel amplification mechanism and utilizing aptamer response type DNA hydrogel to release CAT under the stimulation of the target aptamer 2 O 2 Production of O 2 And O is a 2 And carrying out redox reaction with the oxidation heating material to release a large amount of heat, so as to cause temperature change, thereby constructing a temperature type biosensor to realize high-sensitivity and high-selectivity temperature detection on the target aptamer.
Description
Technical Field
The invention relates to a temperature type biosensor, in particular to a temperature type biosensor constructed based on aptamer response type DNA hydrogel and an oxidation heating material, and also relates to a method for detecting a target aptamer by using the temperature type biosensor, belonging to the technical field of sensing analysis.
Background
Pesticides play a vital role in modern agriculture and aquaculture in order to protect crops from pests and insects. Most agricultural production worldwide relies on the use of Organophosphorus Pesticides (OPs) to increase crop productivity. Compared with other pesticides, malathion is widely applied to agriculture due to the characteristics of high efficiency and low toxicity so as to reduce agricultural production loss. However, excessive use of malathion may cause serious problems in food safety, environment, and human health. In 2013, malathion was reported to be detected in a high out-of-standard concentration in frozen foods of japan-major fisheries company, causing poisoning of consumers. In addition, agricultural products returned in China are billions of dollars per year due to organophosphorus pesticide residues. Malathion belongs to the class 2A carcinogen according to the 2017 carcinogen list published by the world health organization international agency for research on cancer. With the continuous improvement of living standard and the continuous enhancement of environmental protection consciousness of people, the development of a reliable method with high sensitivity and high selectivity for measuring the malathion is imminent.
In recent years, as conventional methods for detection of malathion, there have been mainly electrochemical methods, fluorescence analysis, photoelectrochemical analysis, liquid chromatography and the like ([ 1)]S.Uniyal,R.Sharma.Technological advancement in electrochemical biosensor based detection of organophosphate pesticide chlorpyrifos 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.Carbon dot-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 by butyrylcholinesterase[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 organophosphate pesticide metabolites in large-scale biomonitoring studies[J]Journal of Chromatography a,2016, 1454) however, the practical application of most detection methods is still limited due to the complex instrumentation and the need of professionals. It is crucial to explore a detection strategy that is simple, inexpensive and portable for signal reading devices.
Disclosure of Invention
Aiming at the defects of the detection method of malathion and other target aptamers in the prior art, the invention aims to provide a temperature type biosensor, which is designed based on the specific recognition between an aptamer and the target aptamer, adopts a DNA hydrogel amplification mechanism, and utilizes aptamer response type DNA hydrogel to release CAT under the stimulation of the target aptamer to catalyze H 2 O 2 Production of O 2 And O is a 2 The temperature-type biosensor is constructed by carrying out redox reaction with an oxidation heating material to release a large amount of heat so as to cause temperature change, so that high-sensitivity and high-selectivity temperature detection of a target aptamer is realized.
The second objective of the present invention is to provide a method for detecting a target aptamer by using a temperature biosensor, which can realize high-sensitivity and high-selectivity temperature detection on the target aptamer, and has the advantages of wide detection range and low detection lower limit, wherein the temperature change and the logarithm of the concentration of the target have a good linear relationship.
In order to achieve the above technical objects, the present invention provides a temperature type biosensor comprising an oxygen generating bottle and a heat generating bottle; the bottom of the oxygen generating bottle is provided with aptamer response type DNA hydrogel, and the top of the oxygen generating bottle is provided with a liquid feeder; the bottom of the heat-generating bottle is provided with an oxidation heating material, and the top of the heat-generating bottle is inserted with a thermometer; the middle parts of the oxygen generating bottle and the heat generating bottle are communicated through a conduit; the aptamer response type DNA hydrogel is formed by wrapping catalase in the aptamer crosslinking type DNA hydrogel.
The design principle of the temperature type biosensor provided by the invention is as follows: the DNA sequence is introduced into the hydrophilic polymer chain and can be specifically combined with an aptamer corresponding to a target aptamer, so that the hydrophilic polymer chain can be crosslinked to form a DNA hydrogel with a three-dimensional network structure by utilizing the specific combination between the aptamer and the DNA sequence, meanwhile, CAT is introduced in the crosslinking process, CAT can be embedded or wrapped in the DNA hydrogel in situ, the DNA hydrogel has aptamer response, when the target aptamer exists in the environment where the DNA hydrogel is located, the target aptamer and the aptamer combined with the DNA hydrogel generate a specific reaction, so that the DNA hydrogel is cracked, CAT in the DNA hydrogel is released, the concentration of the target aptamer in the environment is higher, the cracking degree of the DNA hydrogel is higher, the CAT correspondingly released is more, the CAT is in a proportional relation, and the specific catalytic action of the CAT is utilized to realize H 2 O 2 To produce O 2 Generation of O 2 Exothermic reaction with the oxidation exothermic material to cause the ambient temperature to rise, and O is generated 2 The quantity is higher, the exothermic reaction is more violent, the environmental temperature is higher, the quantity and the environmental temperature are in a direct proportion relation, the temperature type biosensor is constructed, and the environmental temperature value is used as a signal, so that the detection of the concentration of the target aptamer in the environment can be realized.
As a preferred embodiment, the aptamer-responsive DNA hydrogel is prepared by the following method: adding catalase into the solution of polyacrylamide containing the grafted DNA chain, uniformly mixing, and adding an aptamer for incubation to obtain the polyacrylamide gel. The polyacrylamide of the grafted DNA strand includes polyacrylamide of the grafted DNA strand-A and polyacrylamide of the grafted DNA strand-B. The DNA strands-A and-B are selected in relation to the corresponding aptamers and can be synthesized according to the design of the selected aptamers, each of the DNA strands-A and-B contains a DNA sequence that can specifically bind to the corresponding aptamer, and the aptamers are determined according to the target aptamers to be assayed (target aptamers such as malathion, proteins, etc.).
As a preferable embodiment, the concentration of catalase in the solution is 200 to 400U, the concentration of polyacrylamide of the graft DNA strand-A in the solution is 0.05 to 0.15mM, and the concentration of polyacrylamide of the graft DNA strand-B in the solution is 0.05 to 0.15mM. The concentration of the aptamer in the solution is 0.05-0.15 mM. The concentrations of the polyacrylamide of the grafted DNA chain-A, the polyacrylamide of the grafted DNA chain-B and the aptamer influence the stability of the DNA hydrogel and the sensitivity of the DNA hydrogel to the response of a target aptamer, if the concentration is too low, the synthesized DNA hydrogel is very sensitive to the response of the target aptamer, but the stability of the DNA hydrogel is extremely low, if the concentration is too high, the synthesized DNA hydrogel is stably improved, but the sensitivity of the synthesized DNA hydrogel to the response of the target aptamer is reduced, and the response is slow, so the concentrations of the polyacrylamide of the grafted DNA chain-A, the polyacrylamide of the grafted DNA chain-B, the aptamer and the like should be controlled to obtain the DNA hydrogel with certain stability and high response sensitivity of the aptamer.
As a preferable scheme, the polyacrylamide of the grafted DNA chain comprises polyacrylamide of grafted DNA chain-A and polyacrylamide of grafted DNA chain-B, wherein the DNA chain-A and the DNA chain-B are DNA chains with different sequences, and the DNA chain-A and the DNA chain-B can be specifically combined with the aptamer. For example, taking as an example the design of a biosensor for detection of malathion, a correspondingly selected malathion aptamer such as 5' -ACTCATCTGTGAATC TCA TCCGTC ACA CCTGCT CTT ATACAAAT TGT TTT TCT CTTAACTTCTTGATCGCT GGT. And the DNA strand-A and the DNA strand-B grafted in polyacrylamide are 5' -acrydite-TTT GAT TCA CAGATGAGT-3' and 5' -acrydite-TTT AGGTGTGACGGATGA-3, respectively. Both DNA strand-A and DNA strand-B are capable of specifically binding to malathion aptamers.
The polyacrylamide for grafting the DNA chain-A and the polyacrylamide for grafting the DNA chain-B are respectively obtained by random copolymerization of the DNA chain-A and the DNA chain-B with acrylamide groups and can be uniformly grafted on the polyacrylamide main chains of the DNA chain-A and the DNA chain-B. Specifically, DNA chain-A and DNA chain-B with acrylamide groups are respectively dispersed in Tris-HCl containing acrylamide, naCl and EDTA, se:Sup>A catalyst is added, and nitrogen bubbling is used for carrying out polymerization reaction, so that polyacrylamide (PS-A) grafted with the DNA chain-A and polyacrylamide (PS-B) grafted with the DNA chain-B are respectively obtained.
As a preferred embodiment, the polyacrylamide of the grafted DNA strand-A and the polyacrylamide of the grafted DNA strand-B are present in an equimolar ratio to the aptamer. The polyacrylamide of the grafted DNA chain-A and the polyacrylamide of the grafted DNA chain-B react with the aptamer according to an equimolar ratio to obtain the more stable three-dimensional DNA hydrogel.
As a preferred embodiment, the incubation conditions are: incubating for 10-30 min at 20-30 ℃. The incubation was carried out under an atmosphere of constantly bubbling nitrogen gas.
As a preferable mode, the oxidation heat-generating material is a composite powder including reduced iron powder, inorganic salt, activated carbon and vermiculite. The oxidation heating material is directly taken from the heating material in the common commercial heating paste in the prior art, the heating material is well known in the prior art, and the components mainly comprise reduced iron powder, inorganic salt, activated carbon and vermiculite. When the oxidation exothermic material is exposed to the air, the powder reacts with O in the air 2 React and release a large amount of heat, wherein the thermochemical equation is 4 Fe(s) +3O 2 (g)→2Fe 2 O 3 (s)；ΔH＝-1648.4kJ·mol -1 Theoretically, under the conditions of constant temperature and pressure, the complete reaction of 0.56g of iron powder with oxygen can release 4.12kJ of thermal energy, and the temperature change caused by the rise of thermal energy can be sensitively detected by using a thermocouple thermometer.
As a preferred option, the thermometer is a thermocouple thermometer, which is chosen for its wide applicability, portability and reliable dosing results.
As a preferred aspect, the catheter is a catheter with a luer two-way valve.
As a preferred embodiment, the liquid feeder is a dropping funnel syringe or the like.
The invention also provides a method for detecting the aptamer by using the temperature type biosensor, which comprises the following steps:
1) Adding the standard aptamer solution into an oxygen generating bottle, incubating the standard aptamer solution and the aptamer response type DNA hydrogel together, and releasing catalase;
2) Adding hydrogen peroxide solution into an oxygen producing bottle, catalyzing hydrogen peroxide to decompose by catalase to generate oxygen, allowing the oxygen to enter the heat producing bottle through a conduit to react with the oxidative heating powder, and recording a temperature value in the heat producing bottle through a thermometer;
3) Repeating the step 1) and the step 2) on a series of standard aptamer solutions with different concentrations to obtain a series of temperature values, and establishing a standard curve of the concentration and the temperature values of the aptamer solutions;
4) And (3) repeating the step 1) and the step 2) on the aptamer solution to be detected to obtain a temperature value, and calculating the concentration of the aptamer solution to be detected according to a standard curve.
As a preferable scheme, after the hydrogen peroxide solution is added into the oxygen generating bottle, the temperature value in the oxygen generating bottle is recorded by a thermometer within 5min.
As a preferable scheme, the oxygen generating bottle and the heat generating bottle of the temperature type biosensor are sealed and vacuumized before detection.
As a preferable scheme, the mass percentage concentration of the hydrogen peroxide is 15-20%. H 2 O 2 The concentration affects the temperature change by affecting the amount and rate of oxygen production, with temperature being a function of H 2 O 2 The concentration increases, but the temperature decreases after the hydrogen peroxide concentration reaches a certain level, which may be attributed to the high concentration of H 2 O 2 Inhibition of CAT catalytic efficiency, detection signal at 20% 2 O 2 The optimum state is reached. Therefore, 20% was regarded as H 2 O 2 The optimum concentration.
As a preferred scheme, the time for incubating the standard aptamer solution and the aptamer-responsive DNA hydrogel is 50-70 min. The reaction time of the aptamer and the aptamer-responsive DNA hydrogel is changed by the temperature change mainly caused by the yield and rate due to the influence of catalase, and the temperature is increased along with the increase of the reaction time and tends to be stable in about 60 min. Therefore, 60min of reaction was selected as the optimum reaction time.
As a preferable scheme, the mass of the heating powder material is 2g 2 O 2 The concentration of (2) is 20%.
The aptamer-responsive DNA hydrogel designed by the invention is aptamer cross-linked DNA waterThe gel is formed by wrapping catalase in situ, when a target aptamer does not exist in the environment, the aptamer cross-linked DNA hydrogel can stably exist, CAT cannot be released into the environment due to the barrier effect of the DNA hydrogel, when the target aptamer exists in the environment, the aptamer in the aptamer cross-linked DNA hydrogel can be specifically combined with the target aptamer, and the aptamer cross-linked DNA hydrogel is cracked due to loss of the aptamer and can release CAT. On the basis, the invention reasonably designs a signal amplification strategy, and further utilizes CAT released from the aptamer crosslinking type DNA hydrogel to amplify H 2 O 2 Conversion to O by catalytic decomposition 2 Generated O 2 Can chemically react with the oxidation exothermic material and release heat, and the temperature in the environment is changed with the temperature of O 2 The content is increased, signals can be read by the portable thermocouple thermometer, the excellent signal amplification effect can be realized, the detection range is widened, the specific combination between the aptamer and the aptamer is utilized, and the selectivity of the sensor is greatly improved.
The construction of the temperature type biosensor comprises the following specific steps:
(1) Preparing an aptamer-responsive DNA hydrogel: 100 μ M of S-A and S-B with an acrylamide group were dispersed in 10mM Tris-HCl buffer (pH 7.4) containing 4% acrylamide, 200mM NaCl,1mM EDTA, respectively, and bubbled with nitrogen gas for 5min to remove air from the solutions; then, 1.4% (v/v) freshly prepared catalyst (0.05gAPS, 25. Mu. LTEMED,0.5mL deionized water) was added to the above mixture to cause polymerization of acrylamide to form se:Sup>A polyacrylamide polymer, followed by nitrogen bubbling for 5min to form polymer chain A (PS-A) and polymer chain B (PS-B); thereafter, CAT (300U) was added thereto, and PS-se:Sup>A and PS-B were mixed with the target aptamer at se:Sup>A 1 molar ratio (both concentrations were 0.1 mM), and nitrogen was bubbled for 5min; finally, the mixed solution was incubated at 25 ℃ for 20min to form an aptamer-responsive DNA hydrogel composed of aptamer-crosslinked DNA hydrogel-encapsulated catalase.
(2) Construction of a temperature sensing platform: the aptamer response type DNA hydrogel is filled into an oxygen generating bottle, the top of the oxygen generating bottle is provided with a liquid feeder (such as an injector), the middle part of the oxygen generating bottle is connected with a heat generating bottle which is provided with an oxidation heating material and is inserted with a thermometer through a conduit with a luer two-way valve, and the oxygen generating bottle is sealed and vacuumized.
(3) Detection of the target aptamer: adding standard target aptamer solutions with different concentrations into an oxygen generating bottle, incubating the standard aptamer solutions with the aptamer response type DNA hydrogel for 1h, adding quantitative hydrogen peroxide, recording temperature change with a portable thermocouple thermometer within 5min, drawing a standard curve according to the temperature value for the concentration of the standard target aptamer solution, performing concentration test by using the target aptamer solution to be tested instead of the standard target aptamer solution, and obtaining a concentration result through the standard curve.
Compared with the prior art, the technical scheme of the invention has the following beneficial technical effects:
1) The invention improves the detection sensitivity of the target aptamer by a signal amplification technology, takes the aptamer crosslinking type DNA hydrogel wrapping CAT as a template, and after the aptamer crosslinking type DNA hydrogel is specifically combined with the target aptamer, the three-dimensional network structure of the aptamer crosslinking type DNA hydrogel is opened, a large amount of CAT is released from the hydrogel, and H is 2 O 2 Production of O under catalysis of enzymes 2 And O is 2 Chemically reacts with the oxidation exothermic material and releases heat, and the temperature in the system is changed with O 2 The increase in the content increases, resulting in an increase in temperature variation.
2) According to the invention, through the specific recognition of the aptamer and the target aptamer, an aptamer cross-linked DNA hydrogel amplification mechanism is adopted, 3D DNA hydrogel is used as a template, and after the 3D DNA hydrogel is specifically combined with the target aptamer, CAT is released to generate O 2 And the temperature change strength is improved, so that the high-sensitivity and high-selectivity temperature detection on the target aptamer is realized.
3) The method for detecting the target aptamer by the temperature type biosensor constructed based on the aptamer response type DNA hydrogel and the simple oxidation thermal reaction has the linear range of 0.1-1000ng mL -1 The detection limit is as low as 0.03ng ml -1 The temperature change and the logarithm of the concentration of the target have good linear relation, and the method has the advantages of wide detection range, low lower limit and the like.
Drawings
FIG. 1 is a schematic diagram of the construction of the temperature-type biosensor and the detection principle of malathion.
FIG. 2 (A) is a graph showing temperature changes at different CAT concentrations; (B) The change in temperature of the DNA hydrogel temperature aptamer sensor in the presence or absence of a target is shown.
FIG. 3 (A) is H 2 O 2 The concentration optimization graph, (B) is a DNA crosslinking concentration optimization graph, and (C) is a time length optimization graph of malathion and hydrogel reaction.
FIG. 4 shows the results of the detection of the standard sample in example 1, A: example 1DNA hydrogel aptamer sensor to different malathions (0.1-1000 ng mL) -1 ) Temperature response of (c); b, corresponding linear relation; c, specific detection of a temperature type biosensor (Mal: malathion, dic: dichlorvos, chl: chlorpyrifos); and D, detecting the stability of the temperature type biosensor.
Detailed Description
The technical solutions of the present invention are further described below by specific implementation examples, but the scope of the present invention should not be limited thereby.
Example 1
(1) 100 μ M S-A and S-B with acrylamide groups were dispersed in 10mM Tris-HCl buffer (pH = 7.4) containing 4% acrylamide, 200mM NaCl,1mM EDTA, respectively, and bubbled with nitrogen gas for 5min to remove air from the solution. Then, 1.4% (v/v) of a freshly prepared catalyst (0.05g APS, 25. Mu. LTEMED,0.5mL deionized water) was added to the above mixture to cause the acrylamide to polymerize and form a polyacrylamide polymer. Then, nitrogen bubbling was performed for 5min to form polymer chain A (PS-A) and polymer chain B (PS-B); thereafter, CAT (300U) was added thereto, and PS-se:Sup>A and PS-B were mixed with malathion aptamer at se:Sup>A 1 molar ratio (both concentrations were 0.1 mM), and nitrogen was bubbled for 5min. Finally, the mixed solution was incubated at 25 ℃ for 20min to form a CAT-coated aptamer-crosslinked 3D DNA hydrogel.
(2) And (2) incubating 10 mu L of target malathion with different concentrations with the 3D DNA hydrogel prepared in the step (1) for 1h to release corresponding CAT, and marking the hydrogel-containing reaction bottle as an oxygen generating bottle. Then, 2g of the "heating powder" in the warm patch was quickly added to the reaction flask with the thermometer inserted(this is labeled as heat generating bottle), then connect the oxygen generating bottle with the heat generating bottle through the conduit with luer two-way valve, seal and vacuumize. Finally, 0.4mL20% H 2 O 2 Fast injection into oxygen generating bottles due to release of CAT, H 2 O 2 Is catalyzed to O 2 。O 2 The "heating powder" is heated and the temperature rises, and the temperature change is recorded by a portable thermocouple thermometer within 5min. Finding that the temperature variation amplitude and the concentration of the malathion have a determined relationship so as to realize sensitive detection of the malathion; and replacing the malathion standard solution with the solution to be detected to perform the detection, and obtaining a concentration result through a standard curve.
Under the same conditions, dichlorvos (Dic) and chlorpyrifos (Chl) are respectively used as target objects, and the selectivity of the method is examined.
The DNA sequences used in example 1 were purchased from [ Biotechnology engineering (Shanghai) Ltd ] as follows:
FIG. 1 is a schematic diagram of the principle of a malathion detection method by a temperature type biosensor constructed based on simple self-heating reaction in target response type DNA hydrogel and a warm patch. FIG. 2 (A) is a graph showing temperature changes at different CAT concentrations; FIG. 2 (B) is a graph showing the temperature change of the DNA hydrogel temperature aptamer sensor in the presence or absence of a target. FIG. 3 (A) is H 2 O 2 The concentration optimization graph, (B) is a DNA crosslinking concentration optimization graph, and (C) is a time length optimization graph of malathion and hydrogel reaction. FIG. 4 shows the results of the detection of the standard samples in example 1, (A) the DNA hydrogel aptamer sensor in example 1 for different malathion (0.1-1000 ng mL) -1 ) The temperature response of (a); (B) the corresponding linear relationship; (C) Specific detection of DNA hydrogel aptamer sensors (Mal: malathion, dic: dichlorvos, chl: chlorpyrifos); and (D) detecting the stability of the DNA hydrogel aptamer sensor. In the range of 0.1-1000ng mL -1 The temperature change has a good linear relationship with the logarithm of the target concentration within the target concentration range of (1) (fig. 4B). Considering the requirement of practicability, IThe method is examined for specificity and selectivity on a specific target object, and Dic and Chl are selected as interference objects to influence corresponding signals of the sensor. The experimental results confirmed that the sensor has substantially no response to different interfering components and a significant response to a specific target, indicating that the sensor has good selectivity and specificity (fig. 4C).
The above description is an example of the present invention, and all equivalent changes and modifications made according to the claims of the present invention should be covered by the present invention.
Claims (7)
1. A method for detecting an aptamer by using a temperature-type biosensor is characterized in that: the method comprises the following steps:
1) Adding the standard aptamer solution into an oxygen generating bottle, incubating the standard aptamer solution and the aptamer response type DNA hydrogel together, and releasing catalase;
2) Adding hydrogen peroxide solution into an oxygen producing bottle, catalyzing hydrogen peroxide to decompose by catalase to generate oxygen, allowing the oxygen to enter the heat producing bottle through a conduit to react with the oxidative heating powder, and recording a temperature value in the heat producing bottle through a thermometer;
3) Repeating the step 1) and the step 2) on a series of standard aptamer solutions with different concentrations to obtain a series of temperature values, and establishing a standard curve of the concentration and the temperature values of the aptamer solutions;
4) Repeating the step 1) and the step 2) on the aptamer solution to be detected to obtain a temperature value, and calculating the concentration of the aptamer solution to be detected according to a standard curve;
the bottom of the oxygen generating bottle is provided with aptamer response type DNA hydrogel, and the top of the oxygen generating bottle is provided with a liquid feeder; the bottom of the heat generating bottle is provided with an oxidation heating material, and the top of the heat generating bottle is inserted with a thermometer; the middle parts of the oxygen generating bottle and the heat generating bottle are communicated through a conduit; the aptamer response type DNA hydrogel is formed by wrapping catalase in the aptamer cross-linked type DNA hydrogel;
the aptamer response type DNA hydrogel is prepared by the following preparation method: adding catalase into a polyacrylamide solution containing a grafted DNA chain, uniformly mixing, and adding an aptamer for incubation to obtain the polyacrylamide gel-like carrier;
the concentration of catalase in the solution is 200-400U,
the concentration of the polyacrylamide of the grafted DNA chain-A in the solution is 0.05-0.15 mM;
the concentration of the polyacrylamide of the grafted DNA chain-B in the solution is 0.05-0.15 mM;
the concentration of the aptamer in the solution is 0.05-0.15 mM.
2. The method for detecting an aptamer according to claim 1, wherein: the polyacrylamide of the grafted DNA chain comprises polyacrylamide of a grafted DNA chain-A and polyacrylamide of a grafted DNA chain-B, wherein the DNA chain-A and the DNA chain-B are DNA chains with different sequences, and both the DNA chain-A and the DNA chain-B can be specifically combined with an aptamer.
3. The method for detecting an aptamer according to claim 1, wherein: the incubation conditions were: incubating for 10-30 min at 20-30 ℃.
4. The method for detecting an aptamer according to claim 1, wherein: the oxidation heating material is composite powder containing reduced iron powder, inorganic salt, activated carbon and vermiculite.
5. The method for detecting an aptamer according to claim 1, wherein: after the hydrogen peroxide solution is added into the oxygen generating bottle, the temperature value in the oxygen generating bottle is recorded by a thermometer within 5min.
6. The method for detecting an aptamer according to claim 5, wherein: the mass percentage concentration of the hydrogen peroxide is 15-20%.
7. The method for detecting an aptamer according to claim 5, wherein: the incubation time of the standard aptamer solution and the aptamer response type DNA hydrogel is 50-70 min.
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