CN105675563A - Rapid quantitative detection method of ***e suitable for field detection - Google Patents
Rapid quantitative detection method of ***e suitable for field detection Download PDFInfo
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- CN105675563A CN105675563A CN201610037309.7A CN201610037309A CN105675563A CN 105675563 A CN105675563 A CN 105675563A CN 201610037309 A CN201610037309 A CN 201610037309A CN 105675563 A CN105675563 A CN 105675563A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6432—Quenching
Abstract
The invention discloses a rapid quantitative detection method of ***e suitable for field detection, comprising the following steps: (1), preparing a paper base detector: using paper as a detection base, outlining a hydrophilic detection area on the surface of the paper by using a hydrophobic barrier, and covalently coupling water-soluble up-conversion fluorescent nano-material and aptamer ACA-1; (2), surface modifying gold nanoparticles with aptamer ACA-2 to obtain solution AuNPs-ACA-2; (3), deeply optimizing the solution AuNPs-ACA-2; (4), preparing a standard solution of ***e; (5), drawing a standard curve for ***e detection; (6) measuring ***e concentration in bodily fluid to be detected.The unsolved problem with rapid quantitative detection in the field of ***e detection is solved, rapid, quantitative and highly sensitive detection of ***e concentration in a bodily fluid sample is implemented, and quantitative detection sensitivity is up to 0.05 MuMu.
Description
Technical field
The invention belongs to ***e Quantitative detection technical field, be specifically related to the paper substrate detection method of a kind of detection ***e quick, quantitative, highly sensitive based on upconversion fluorescence nano material.
Background technology
Drug abuse is one of social problem that the harm mankind are the most serious at present, and ***e is most widely used one in all kinds of drugs. Human body can be produced very big threat by ***e abuse so that human heart rate is accelerated, rapid breathing, even occurs trembling, the phenomenon such as spasm, convulsions, and human security, social stability and world peace are constituted grave danger. Therefore, study novel, sensitive, quantitative ***e detection method, for the prohibition of drug, poisonous substance detection, criminal identification etc., there is very important practice significance and using value.
The traditional ***e detection method reported at present has: Optical Analysis Method, electrochemical methods, high performance capillary electrophoresis, nuclear magnetic resonance spectrometry etc., these traditional detection methods exist that detection sensitivity is inadequate, complicated operation is consuming time, rely on accurate expensive instrument, need the shortcomings such as professional is operated, and usually can not meet the requirement researched and analysed with Clinical detection. Field quick detection drugs method based on monoclonal antibody is most widely used, but such method has some limitations, as antibody self-defect, antibody preparation work amount non-specific binding big, separate sources antibody produce false positive signal etc. In recent years, drugs field fast detection method based on aptamer is necessarily developed, and the detection method specificity based on aptamer is good, and sensitivity is higher relative to antibody test, but order great majority are also limited to the laboratory research stage, are not suitable for for Site Detection.
In sum, this area still lacks the gratifying ***e fast quantitative measurement method for detecting suitable in Site Detection, therefore, in the urgent need to developing the ***e onthe technology of site test with the advantage such as sensitive, quick, accurate, specificity is good.
Summary of the invention
The technical problem to be solved is for overcoming the deficiencies in the prior art, it is provided that plant a kind of ***e fast quantitative measurement method for detecting suitable in Site Detection, it is possible to directly obtain ***e detection by quantitative result, it is not necessary to secondary detection.
The present invention solves that the technical scheme that above-mentioned technical problem provides is:
A kind of ***e fast quantitative measurement method for detecting suitable in Site Detection, comprises the following steps:
(1) paper substrate detecting device is prepared: choose paper as detection substrate, hydrophilic detection region is drawn a circle to approve as hydrophobic barrier at paper surface using water-insoluble solvent, by water-soluble upconversion fluorescence nano material and single-chain nucleic acid aptamers ACA-1 covalent coupling in described hydrophilic detection region, obtain paper substrate detecting device;
(2) choosing gold nano grain (AuNPs) and at its surface markers single-chain nucleic acid aptamers ACA-2, products therefrom is denoted as AuNPs-ACA-2, is scattered in buffer, obtains AuNPs-ACA-2 solution;
(3) concentration optimization of AuNPs-ACA-2 solution: add the ***e solution that volume is V1 fixed concentration and the AuNPs-ACA-2 solution that volume is V2 variable concentrations respectively in the hydrophilic detection region of the paper substrate detecting device described in step (1), remove unreacted AuNPs-ACA-2 after having reacted; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region and read green intensity F in fluoroscopic image with software; When setting the concentration of AuNPs-ACA-2 solution as 0, gained green intensity blank sample intensity is designated as F0, calculate the fluorescent quenching efficiency (F corresponding to variable concentrations AuNPs-ACA-2 solution0-F)/F0, obtain fluorescent quenching efficiency maximum time corresponding AuNPs-ACA-2 solution concentration C;
(4) standard solution of ***e is prepared: prepare the ***e solution of variable concentrations with body fluid, be ***e standard solution;
(5) ***e examination criteria curve is drawn: in the hydrophilic detection region of the paper substrate detecting device described in step (1), add volume be respectively the ***e standard solution of V1 and the AuNPs-ACA-2 solution of volume to be V2 concentration be C, after having reacted, remove unreacted AuNPs-ACA-2; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region and read green intensity F ' in fluoroscopic image with software; When setting the concentration of ***e standard solution as 0, gained green intensity blank sample intensity is designated as F0', with fluorescent quenching efficiency (F0’-F’)/F0' for vertical coordinate, with the logarithm value of ***e concentration of standard solution for abscissa drawing standard curve;
(6) mensuration of ***e concentration in body fluid to be measured: add volume respectively in the hydrophilic detection region of the paper substrate detecting device described in step (1) and be the body fluid to be measured of V1 and the AuNPs-ACA-2 solution of volume to be V2 concentration be C, remove unreacted AuNPs-ACA-2 after having reacted; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region and read green intensity Fx in fluoroscopic image with software; Calculate fluorescent quenching efficiency (F0’-Fx)/F0', substitute into standard curve and obtain the ***e concentration in body fluid to be measured.
By such scheme, the shape circular in described hydrophilic detection region, the preferred 2-6mm of internal diameter.
By such scheme, described water-soluble upconversion fluorescence nano material pattern is spherical, and particle diameter is at 30-100nm, and finishing has amino. Wherein, the substrate of described upconversion fluorescence nano material is upconversion fluorescence nano material NaYF4、NaGdF4Deng, amido modified polymine, aminoethyl phosphonic acid etc. can be adopted as modifying agent, to realize water solublity.
By such scheme, in step (1) during covalent coupling, water-soluble upconversion fluorescence nano material is configured to the solution that concentration is 0.2-0.8mg/ml, single-chain nucleic acid aptamers ACA-1 is configured to the solution that concentration is 0.5-4 μM, both are 1.3-12 μ L at addition, and cumulative volume is the summation of V1 and V2; Or it is configured to both mixed solutions, adjusts addition in right amount.
By such scheme, described gold nano grain pattern is spherical, and particle diameter is at 10-30nm, and finishing has citrate.
By such scheme, described single-chain nucleic acid ACA-1 and single-chain nucleic acid ACA-2, its sequence is respectively as follows: 5 '-NH2-TTTTTCAAGAACTGAGGG-3 ' and 5 '-SH-TTTTTACAGCAGGGTGAAGTAACTTCTTG-3 '.
By such scheme, described paper is required to be hydrophilic paper, including printing paper, ordinary filter paper, cellulosic filter paper, nitrocellulose filter etc., it is preferred to cellulosic filter paper.
By such scheme, the water-insoluble solvent in step (1) is selected from hydrophobic inks, paraffin, photoresist, alkyl ketene dimer etc.
By such scheme, the pH of buffer scope described in step (2) is at 7.0-7.5, it is possible to select the 20mMTris-HCl buffer solution containing 100mMNaCl; In step (3), buffer is identical with step (2).
By such scheme, the preparation steps of the ***e solution in step (3) be after solvent in ***e standard substance is volatilized completely add buffer (this buffer can select: the 20mMTris-HCl containing 100mMNaCl, pH is the buffer of 7.4), ice-bath ultrasonic 30min obtains ***e storing solution, then by same buffer, storing solution is diluted to certain concentration. The certain concentration of this ***e solution can set that to be 10 μMs, and the concentration range of AuNPs-ACA-2 solution is generally 0-0.305 μM.
By such scheme, with buffer solution to remove unreacted AuNPs-ACA-2 in step (3). This buffer is the 20mMTris-HCl buffer containing 100mMNaCl, and the volumn concentration of wherein Polyethylene Glycol (PEG) is 1%-2%, the volumn concentration of polysorbas20 (Tween20) is 0.01%-0.05%, and the pH of this buffer ranges for 7.0-7.5; Described washing concretely comprises the following steps: is first inverted by paper substrate detecting device, makes hydrophilic detection region down, adds 5-10 μ L lavation buffer solution from above, and promotes solution flowing, repeated washing 3-5 time in lower section pad absorbent paper.
By such scheme, in described step (4), the concentration range of ***e standard solution is 0-50 μM.
By square being directly proportional of such scheme, described step (3) and the volume V1 in step (5), (6) and volume V2 and hydrophilic detection regional diameter. When hydrophilic detection region is internal diameter 2-6mm circle, volume V1 and volume V2 is 1.3-12 μ L all preferably.
Being 20-37 DEG C by such scheme, described step (3) and the reaction temperature in step (5), (6), the response time is reaction 20-60min.
By such scheme, described step (3) and mobile phone can be adopted in step (5), (6) to shoot hydrophilic detection region fluoroscopic image, reads green intensity in fluoroscopic image with mobile phone RGB software.
By such scheme, described body fluid specifically includes blood, saliva, tear, urine, perspiration etc.
Principles of the invention is: the specific recognition function of FRET (fluorescence resonance energy transfer) and ***e aptamers combined, it is achieved ***e detection by quantitative. Cocaine aptamers is divided into two independent DNA (ACA-1 and ACA-2) by the present invention, and the two does not interact when being absent from ***e, and generates the secondary structure of aptamers when ***e exists.Upconversion fluorescence nano material and ACA-1 covalency as fluorogenic donor is fixed on note detection region by the present invention, gold nano grain finishing ACA-2 as fluorescent receptor, when in body fluid to be measured containing ***e, ACA-1 and ACA-2 and ***e react formation aptamers secondary structure (Fig. 1), AuNPs is fixed on paper surface, distance between upconversion fluorescence nano material and AuNPS is furthered, in FRET (fluorescence resonance energy transfer) distance range (1-10nm), AuNPs can the fluorescence of effectively quencher upconversion fluorescence nano material, there is dependency in fluorescent quenching efficiency and ***e concentration, the paper substrate detecting device reacted is placed in detection device (Fig. 2) and shoots fluorescence picture with mobile phone, dependency is there is between fluorescence picture color and ***e concentration, the present invention utilizes this principle to realize the detection by quantitative of ***e.
Compared with prior art, the invention has the beneficial effects as follows:
1, the present invention directly can with the naked eye carry out rational judgment according to green intensity, mobile phone can also be utilized as signal collection device simultaneously, directly obtain ***e detection by quantitative result, without secondary detection, and Site Detection sensitivity can be suitable with test in laboratory method, far above existing in-situ check and test method;
2, the aptamer appropriate design of specific recognition ***e is become two sections of sequences by the present invention, it is to avoid the interference of the secondary structure of aptamers own, reduces background signal, improves and analyzes detection sensitivity;
3, up-conversion fluorescence resonance energy is combined by the present invention with aptamers specific recognition capability, by optimizing the relative concentration of upconversion fluorescence nano material and aptamers, it is effectively improved the fluorescent quenching efficiency of system, it is effectively increased detection sensitivity, qualitative detection sensitivity is up to 0.05 μM, and detection by quantitative sensitivity is up to 0.5 μM.
4, the present invention utilize upconversion fluorescence nano material near infrared light to excite, the advantage of VISIBLE LIGHT EMISSION, directly detect in complicated humoral sample, it is not necessary to early stage pre-treatment step, simple to operate, be more applicable for Site Detection.
Accompanying drawing explanation
Fig. 1 is the Cleaning Principle figure of a kind of ***e fast quantitative measurement method for detecting suitable in Site Detection.
Fig. 2 is the detection device model of a kind of ***e fast quantitative measurement method for detecting suitable in Site Detection.
Fig. 3 is AuNPs-ACA-2 concentration and fluorescent quenching relationship between efficiency figure in embodiment 1.
Fig. 4 is ***e concentration and fluorescent quenching relationship between efficiency figure in saliva in embodiment 1.
Detailed description of the invention
In order to make those skilled in the art be more fully understood that technical scheme, below in conjunction with specific embodiment, the present invention is described in further detail.
1, in following embodiment, described water-soluble upconversion fluorescence nano material (NaYF4: Yb, Er, wherein Y, Yb, Er mol ratio is 80:18:2, is denoted as UCPs) concrete preparation method is: select finishing to have the spherical upconversion fluorescence nano material NaYF of oleic acid4: Yb, Er (are denoted as OA-UCPs), are dispersed in chloroform, obtain the OA-UCPs chloroformic solution that concentration is 1 μm of ol/L; 30ml diglycol and 600mg polymine are joined in 100mL there-necked flask; heat under magnetic agitation and argon shield after system evacuation to 110 DEG C; 1h is reacted after being added dropwise over by 1mLOA-UCPs chloroformic solution; then heat to 240 DEG C and continue reaction 6h; it is cooled to flask adds equal-volume ethanol after room temperature until system, centrifugal is precipitated, be scattered in pure water by after precipitation second alcohol and water centrifuge washing 3 times; namely obtain upconversion fluorescence nano material, be denoted as PEI-UCPs.This upconversion fluorescence nano material pattern is spherical, and particle diameter is at 30-100nm, and finishing has amino.
Certainly, as long as water miscible upconversion fluorescence nano material all can be suitably used for the present invention.
2, in following embodiment, the preparation of described gold nano grain and surface markers thereof carry out as follows:
(1) trisodium citrate reduction gold chloride method is adopted to prepare gold nano grain: gold chloride is first made into the aqueous solution that mass fraction is 2%, take 2mL and and add the heating of 50mL high purity water to seething with excitement in round-bottomed flask, add the trisodium citrate aqueous solution of 5mL38.8mM with vigorous stirring, continuing heated and boiled 15min, flaxen aqueous solution of chloraurate becomes claret; Naturally cool to after room temperature after with 0.22 μm of membrane filtration until system, namely obtain the gold nano grain aqueous solution that concentration is 3.7 μm of ol/L, be placed in 4 DEG C of refrigerators standby. This gold nano grain pattern is spherical, and particle diameter is at 10-30nm, and finishing has citrate.
(2) surface markers: after being mixed by 1nmol single-chain nucleic acid aptamers ACA-2 and 5nmol three (2-carboxyethyl) phosphine (TCEP), room temperature reaction 1h is to open the disulfide bond of ACA-2; Having reacted rear solution and 1.5mL3.7 μm of ol/L gold nano grain solution mixes and is incorporated in room temperature reaction 16h, then progressively regulating solution salt concentration with 2mol/LNaCl solution is 0.1mol/L, is placed in room temperature and again reacts 24h; React rear products therefrom high purity water centrifuge washing three times, be then dispersed in Tris-HCl buffer and save backup, be AuNPs-ACA-2 solution. The wherein 20mMTris-HCl buffer consisted of containing 100mMNaCl of buffer, pH ranges for 7.0-7.5.
3, in the present invention, prepare paper substrate detecting device: choose paper as detection substrate, hydrophilic detection region is drawn a circle to approve as hydrophobic barrier at paper surface using water-insoluble solvent, by water-soluble upconversion fluorescence nano material and single-chain nucleic acid aptamers ACA-1 covalent coupling in described hydrophilic detection region, paper substrate detecting device can be obtained.
(1) paper substrate described in following example is prepared as follows: draws detection zone region pattern on computers and prints at paper surface by normal printer; The plastic formwork identical with hydrophilic detection zone region pattern shape is covered paper substrate surface, with oil pen along detection zone region pattern edge draw ink line, draw and be placed on room temperature and stand and make the solvent in the hydrophobic oil line of ink marker volatilize completely, remained in the hydrophobic barrier of formation such as resin paper within, Organic substance.
(2) water-soluble upconversion fluorescence nano material and single-chain nucleic acid ACA-1 covalent coupling are prepared as follows by following example at paper surface: preparation NaIO4Concentration is 26mM, LiCl concentration is the mixed aqueous solution of 47mM, is immersed in 75mL mixed aqueous solution by 0.5g (really in mass) paper substrate and reacts 1h in 55 DEG C; Then the washing of immersion gained paper substrate is placed on for 2 times in baking oven and dries, the mixed solution of the 2.6-24 μ L summation of V1 and V2 (volume here be) PEI-UCPs and ACA-1 is added subsequently (in this mixed solution in each hydrophilic detection region, PEI-UCPs concentration is 0.5mg/ml, ACA-1 concentration is 0.5-4 μM, sodium cyanoborohydride NaCNBH3Concentration is 200mM, and solvent is 100mMHEPES buffering, and pH value of solution is 7.2), and in room temperature reaction 30min; React polysorbas20 (Tween20) the solution soaking washing 5min that rear paper substrate volume fraction is 0.02%, dry after washing twice, it is paper substrate detecting device, saves backup.
Embodiment 1
A kind of ***e fast quantitative measurement method for detecting suitable in Site Detection, comprises the following steps:
(1) paper substrate detecting device is prepared as previously mentioned, wherein cellulosic filter paper selected by paper, circular hydrophilic detection region internal diameter is 3mm, adding the mixed solution of 6 μ LPEI-UCPs and ACA-1 in each hydrophilic detection region, wherein water-soluble upconversion fluorescence nano material PEI-UCPs concentration is 0.5mg/mL and single-chain nucleic acid ACA-1 concentration is 2 μMs;
(2) AuNPs-ACA-2 solution is prepared as previously mentioned;
(3) concentration optimization of AuNPs-ACA-2 solution: add the AuNPs-ACA-2 solution (concentration respectively 0 of the ***e solution of 3 μ L10 μM and 3 μ L variable concentrations respectively in the hydrophilic detection region of the paper substrate detecting device described in step (1), 0.07,0.105,0.14,0.175,0.21,0.28,0.315 μM), under 25 DEG C of conditions, react 40min, remove unreacted AuNPs-ACA-2 5 times with 6 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity F in fluoroscopic image with mobile phone RGB software; When setting the concentration of AuNPs-ACA-2 solution as 0, gained green intensity blank sample intensity is designated as F0, calculate the fluorescent quenching efficiency (F corresponding to variable concentrations AuNPs-ACA-2 solution0-F)/F0, obtaining AuNPs-ACA-2 solution concentration C corresponding during fluorescent quenching efficiency maximum (54.4%) is 0.28 μM;
(4) standard solution of ***e is prepared: with saliva dilution ***e stock solution, the ***e solution of preparation variable concentrations, be ***e standard solution, concentration respectively 0,0.05,0.1,0.5,1,5,10,50 μM;
(5) ***e examination criteria curve is drawn: in the hydrophilic detection region of the paper substrate detecting device described in step (1), add the ***e standard solution of 3 μ L and the AuNPs-ACA-2 solution of 3 μ L concentration 0.28 μM respectively, under 25 DEG C of conditions, react 40min, remove unreacted AuNPs-ACA-2 5 times with 6 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity F ' in fluoroscopic image with mobile phone RGB software; When setting the concentration of ***e standard solution as 0, gained green intensity blank sample intensity is designated as F0', with fluorescent quenching efficiency (F0’-F’)/F0' for vertical coordinate, with the logarithm value of ***e concentration of standard solution for abscissa drawing standard curve, standard curve equation is Y=0.18X+0.60;
(6) mensuration of ***e concentration in saliva to be measured: add the body fluid to be measured of 3 μ L and the AuNPs-ACA-2 solution of 3 μ L concentration 0.28 μM respectively in the hydrophilic detection region of the paper substrate detecting device described in step (1), under 25 DEG C of conditions, react 40min, remove unreacted AuNPs-ACA-2 5 times with 6 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity Fx in fluoroscopic image with mobile phone RGB software; Calculate fluorescent quenching efficiency (F0’-Fx)/F0', it is 0.463 μM that substitution standard curve obtains the ***e concentration in body fluid to be measured.
In known body fluid to be measured, ***e concentration is 0.5 μM, and the testing result accuracy rate of the present invention can reach 92.6%, and the standard deviation of three repeated measure is 11%, and accuracy and the repeatability of testing result are good.
As shown in Figure 3: in the detection method that the present invention sets up, the concentration of AuNPs-ACA-2 affects fluorescent quenching efficiency within the specific limits, and AuNPs-ACA-2 concentration is more big, and fluorescent quenching efficiency is more high.
As shown in Figure 4: the detection method that the present invention sets up can realize ***e detection within the scope of finite concentration, ***e concentration is more big, the green-emitting fluorescent intensity of fluoroscopic image is more weak, fluorescent quenching efficiency and ***e log concentration value present good linear relationship, showing that this detection method can realize the qualitative detection to saliva ***e and detection by quantitative, its sensitivity is respectively up to 0.05 μM and 0.5 μM.
As shown in Table 1: for the saliva sample of ***e of variable concentrations, the accuracy rate of the detection method testing result that the present invention sets up is at 88.8%-103.2%, relative standard deviation between repeated detection result is less than 15%, it was shown that the detection method that the present invention sets up has good Stability and veracity.
Cocaine detection recovery testu in table 1 saliva sample
| Sample number into spectrum | Add concentration (μM) | Test concentrations (μM) | The response rate (%) | Relative standard deviation (%) |
| 1 | 0.05 | 0.0516 | 103.2 | 14.6 |
| 2 | 0.5 | 0.463 | 92.6 | 11.0 |
| 3 | 2 | 1.775 | 88.8 | 8.8 |
Embodiment 2
A kind of ***e fast quantitative measurement method for detecting suitable in Site Detection, comprises the following steps:
(1) paper substrate detecting device is prepared as previously mentioned, wherein cellulosic filter paper selected by paper, circular hydrophilic detection region internal diameter is 2mm, adding the mixed solution of 2.6 μ LPEI-UCPs and ACA-1 in each hydrophilic detection region, wherein water-soluble upconversion fluorescence nano material concentration is 0.5mg/mL and single-chain nucleic acid ACA-1 concentration is 0.5 μM;
(2) AuNPs-ACA-2 solution is prepared as previously mentioned;
(3) concentration optimization of AuNPs-ACA-2 solution: add the AuNPs-ACA-2 solution (concentration respectively 0 of the ***e solution of 1.3 μ L10 μM and 1.3 μ L variable concentrations respectively in the hydrophilic detection region of the paper substrate detecting device described in step (1), 0.07,0.105,0.14,0.175,0.21,0.28,0.315 μM), under 20 DEG C of conditions, react 20min, remove unreacted AuNPs-ACA-2 3 times with 3 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity F in fluoroscopic image with mobile phone RGB software; When setting the concentration of AuNPs-ACA-2 solution as 0, gained green intensity blank sample intensity is designated as F0, calculate the fluorescent quenching efficiency (F corresponding to variable concentrations AuNPs-ACA-2 solution0-F)/F0, obtain fluorescent quenching efficiency maximum time corresponding AuNPs-ACA-2 solution concentration C be 0.14 μM;
(4) standard solution of ***e is prepared: employment serum-dilution ***e stock solution, the ***e solution of preparation variable concentrations, it is ***e standard solution, concentration respectively 0,0.05,0.1,0.5,1,5,10,50 μM;
(5) ***e examination criteria curve is drawn: in the hydrophilic detection region of the paper substrate detecting device described in step (1), add the ***e standard solution of 1.3 μ L and the AuNPs-ACA-2 solution of 1.3 μ L concentration 0.14 μM respectively, under 20 DEG C of conditions, react 20min, remove unreacted AuNPs-ACA-2 3 times with 3 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity F ' in fluoroscopic image with mobile phone RGB software;When setting the concentration of ***e standard solution as 0, gained green intensity blank sample intensity is designated as F0', with fluorescent quenching efficiency (F0’-F’)/F0' for vertical coordinate, with the logarithm value of ***e concentration of standard solution for abscissa drawing standard curve, standard curve equation is Y=0.11X+0.43;
(6) mensuration of ***e concentration in test serum: add the body fluid to be measured of 1.3 μ L and the AuNPs-ACA-2 solution of 1.3 μ L concentration 0.14 μM respectively in the hydrophilic detection region of the paper substrate detecting device described in step (1), under 20 DEG C of conditions, react 20min, remove unreacted AuNPs-ACA-2 3 times with 3 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity Fx in fluoroscopic image with mobile phone RGB software; Calculate fluorescent quenching efficiency (F0’-Fx)/F0', it is 0.185 μM that substitution standard curve obtains the ***e concentration in body fluid to be measured.
In known test serum, ***e concentration is 0.2 μM, and the testing result accuracy rate of the present invention can reach 92.5%, and the standard deviation of three repeated measure is 9.8%, and accuracy and the repeatability of testing result are good.
Embodiment 3
A kind of ***e fast quantitative measurement method for detecting suitable in Site Detection, comprises the following steps:
(1) paper substrate detecting device is prepared as previously mentioned, wherein cellulosic filter paper selected by paper, circular hydrophilic detection region internal diameter is 6mm, adding the mixed solution of 24 μ LPEI-UCPs and ACA-1 in each hydrophilic detection region, wherein water-soluble upconversion fluorescence nano material concentration is 0.5mg/mL and single-chain nucleic acid ACA-1 concentration is 4 μMs;
(2) AuNPs-ACA-2 solution is prepared as previously mentioned;
(3) concentration optimization of AuNPs-ACA-2 solution: add the AuNPs-ACA-2 solution (concentration respectively 0 of the ***e solution of 12 μ L10 μM and 12 μ L variable concentrations respectively in the hydrophilic detection region of the paper substrate detecting device described in step (1), 0.07,0.105,0.14,0.175,0.21,0.28,0.315 μM), under 37 DEG C of conditions, react 40min, remove unreacted AuNPs-ACA-2 3 times with 24 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity F in fluoroscopic image with mobile phone RGB software; When setting the concentration of AuNPs-ACA-2 solution as 0, gained green intensity blank sample intensity is designated as F0, calculate the fluorescent quenching efficiency (F corresponding to variable concentrations AuNPs-ACA-2 solution0-F)/F0, obtain fluorescent quenching efficiency maximum time corresponding AuNPs-ACA-2 solution concentration C be 0.315 μM;
(4) standard solution of ***e is prepared: dilute ***e stock solution, the ***e solution of preparation variable concentrations with tear, be ***e standard solution, concentration respectively 0,0.05,0.1,0.5,1,5,10,50 μM;
(5) ***e examination criteria curve is drawn: in the hydrophilic detection region of the paper substrate detecting device described in step (1), add the ***e standard solution of 12 μ L and the AuNPs-ACA-2 solution of 12 μ L concentration 0.315 μM respectively, under 37 DEG C of conditions, react 40min, remove unreacted AuNPs-ACA-2 3 times with 24 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity F ' in fluoroscopic image with mobile phone RGB software;When setting the concentration of ***e standard solution as 0, gained green intensity blank sample intensity is designated as F0', with fluorescent quenching efficiency (F0’-F’)/F0' for vertical coordinate, with the logarithm value of ***e concentration of standard solution for abscissa drawing standard curve, standard curve equation is Y=0.21X+0.55;
(6) mensuration of ***e concentration in tear to be measured: add the body fluid to be measured of 12 μ L and the AuNPs-ACA-2 solution of 12 μ L concentration 0.315 μM respectively in the hydrophilic detection region of the paper substrate detecting device described in step (1), under 37 DEG C of conditions, react 40min, remove unreacted AuNPs-ACA-2 3 times with 24 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity Fx in fluoroscopic image with mobile phone RGB software; Calculate fluorescent quenching efficiency (F0’-Fx)/F0', it is 11.025 μMs that substitution standard curve obtains the ***e concentration in body fluid to be measured.
In known tear to be measured, ***e concentration is 10 μMs, and the testing result accuracy rate of the present invention can reach 110.25%, and the standard deviation of three repeated measure is 8.3%, and accuracy and the repeatability of testing result are good.
Embodiment 4
A kind of ***e fast quantitative measurement method for detecting suitable in Site Detection, comprises the following steps:
(1) paper substrate detecting device is prepared as previously mentioned, wherein cellulosic filter paper selected by paper, circular hydrophilic detection region internal diameter is 4mm, adding the mixed solution of 10 μ LPEI-UCPs and ACA-1 in each hydrophilic detection region, wherein water-soluble upconversion fluorescence nano material concentration is 0.5mg/mL and single-chain nucleic acid ACA-1 concentration is 1 μM;
(2) AuNPs-ACA-2 solution is prepared as previously mentioned;
(3) concentration optimization of AuNPs-ACA-2 solution: add the AuNPs-ACA-2 solution (concentration respectively 0 of the ***e solution of 5 μ L10 μM and 5 μ L variable concentrations respectively in the hydrophilic detection region of the paper substrate detecting device described in step (1), 0.07,0.105,0.14,0.175,0.21,0.28,0.315 μM), under 30 DEG C of conditions, react 60min, remove unreacted AuNPs-ACA-2 5 times with 10 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity F in fluoroscopic image with mobile phone RGB software; When setting the concentration of AuNPs-ACA-2 solution as 0, gained green intensity blank sample intensity is designated as F0, calculate the fluorescent quenching efficiency (F corresponding to variable concentrations AuNPs-ACA-2 solution0-F)/F0, obtain fluorescent quenching efficiency maximum time corresponding AuNPs-ACA-2 solution concentration C be 0.28 μM;
(4) standard solution of ***e is prepared: dilute ***e stock solution, the ***e solution of preparation variable concentrations by urine, be ***e standard solution, concentration respectively 0,0.05,0.1,0.5,1,5,10,50 μM;
(5) ***e examination criteria curve is drawn: in the hydrophilic detection region of the paper substrate detecting device described in step (1), add the ***e standard solution of 5 μ L and the AuNPs-ACA-2 solution of 5 μ L concentration 0.28 μM respectively, under 30 DEG C of conditions, react 60min, remove unreacted AuNPs-ACA-2 5 times with 10 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity F ' in fluoroscopic image with mobile phone RGB software;When setting the concentration of ***e standard solution as 0, gained green intensity blank sample intensity is designated as F0', with fluorescent quenching efficiency (F0’-F’)/F0' for vertical coordinate, with the logarithm value of ***e concentration of standard solution for abscissa drawing standard curve, standard curve equation is Y=0.15X+0.57;
(6) mensuration of ***e concentration in urine to be measured: add the body fluid to be measured of 5 μ L and the AuNPs-ACA-2 solution of 5 μ L concentration 0.28 μM respectively in the hydrophilic detection region of the paper substrate detecting device described in step (1), under 30 DEG C of conditions, react 60min, remove unreacted AuNPs-ACA-2 5 times with 10 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity Fx in fluoroscopic image with mobile phone RGB software; Calculate fluorescent quenching efficiency (F0’-Fx)/F0', it is 26.783 μMs that substitution standard curve obtains the ***e concentration in body fluid to be measured.
In known urine to be measured, ***e concentration is 25 μMs, and the testing result accuracy rate of the present invention can reach 107.13%, and the standard deviation of three repeated measure is 6.9%, and accuracy and the repeatability of testing result are good.
Embodiment 5
A kind of ***e fast quantitative measurement method for detecting suitable in Site Detection, comprises the following steps:
(1) paper substrate detecting device is prepared as previously mentioned, wherein cellulosic filter paper selected by paper, circular hydrophilic detection region internal diameter is 5mm, adding the mixed solution of 16 μ LPEI-UCPs and ACA-1 in each hydrophilic detection region, wherein water-soluble upconversion fluorescence nano material concentration is 0.5mg/mL and single-chain nucleic acid ACA-1 concentration is 2 μMs;
(2) AuNPs-ACA-2 solution is prepared as previously mentioned;
(3) concentration optimization of AuNPs-ACA-2 solution: add the AuNPs-ACA-2 solution (concentration respectively 0 of the ***e solution of 8 μ L10 μM and 8 μ L variable concentrations respectively in the hydrophilic detection region of the paper substrate detecting device described in step (1), 0.07,0.105,0.14,0.175,0.21,0.28,0.315 μM), under 37 DEG C of conditions, react 40min, remove unreacted AuNPs-ACA-2 5 times with 16 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity F in fluoroscopic image with mobile phone RGB software; When setting the concentration of AuNPs-ACA-2 solution as 0, gained green intensity blank sample intensity is designated as F0, calculate the fluorescent quenching efficiency (F corresponding to variable concentrations AuNPs-ACA-2 solution0-F)/F0, obtain fluorescent quenching efficiency maximum time corresponding AuNPs-ACA-2 solution concentration C be 0.21 μM;
(4) standard solution of ***e is prepared: dilute ***e stock solution, the ***e solution of preparation variable concentrations with perspiration, be ***e standard solution, concentration respectively 0,0.05,0.1,0.5,1,5,10,50 μM;
(5) ***e examination criteria curve is drawn: in the hydrophilic detection region of the paper substrate detecting device described in step (1), add the ***e standard solution of 8 μ L and the AuNPs-ACA-2 solution of 8 μ L concentration 0.21 μM respectively, under 37 DEG C of conditions, react 40min, remove unreacted AuNPs-ACA-2 5 times with 16 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity F ' in fluoroscopic image with mobile phone RGB software;When setting the concentration of ***e standard solution as 0, gained green intensity blank sample intensity is designated as F0', with fluorescent quenching efficiency (F0’-F’)/F0' for vertical coordinate, with the logarithm value of ***e concentration of standard solution for abscissa drawing standard curve, standard curve equation is Y=0.169X+0.63;
(6) mensuration of ***e concentration in perspiration to be measured: add the body fluid to be measured of 8 μ L and the AuNPs-ACA-2 solution of 8 μ L concentration 0.21 μM respectively in the hydrophilic detection region of the paper substrate detecting device described in step (1), under 37 DEG C of conditions, react 40min, remove unreacted AuNPs-ACA-2 5 times with 16 μ L lavation buffer solution washings; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region with mobile phone and read green intensity Fx in fluoroscopic image with mobile phone RGB software; Calculate fluorescent quenching efficiency (F0’-Fx)/F0', it is 0.046 μM that substitution standard curve obtains the ***e concentration in body fluid to be measured.
In known body fluid to be measured, ***e concentration is 0.05 μM, and the testing result accuracy rate of the present invention can reach 92%, and the standard deviation of three repeated measure is 12.8%, and accuracy and the repeatability of testing result are good.
The above is only the preferred embodiment of the present invention, it is noted that for the person of ordinary skill of the art, without departing from the concept of the premise of the invention, it is also possible to making some improvement and conversion, these broadly fall into protection scope of the present invention.
Claims (10)
1. the ***e fast quantitative measurement method for detecting being applicable to Site Detection, it is characterised in that comprise the following steps:
(1) paper substrate detecting device is prepared: choose paper as detection substrate, hydrophilic detection region is drawn a circle to approve as hydrophobic barrier at paper surface using water-insoluble solvent, by water-soluble upconversion fluorescence nano material and single-chain nucleic acid aptamers ACA-1 covalent coupling in described hydrophilic detection region, obtain paper substrate detecting device;
(2) choosing gold nano grain and at its finishing single-chain nucleic acid aptamers ACA-2, products therefrom is denoted as AuNPs-ACA-2, is scattered in buffer solution, obtains AuNPs-ACA-2 solution;
(3) concentration optimization of AuNPs-ACA-2 solution: add the ***e solution that volume is V1 fixed concentration and the AuNPs-ACA-2 solution that volume is V2 variable concentrations respectively in the hydrophilic detection region of the paper substrate detecting device described in step (1), remove unreacted AuNPs-ACA-2 after having reacted; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region and read green intensity F in fluoroscopic image with software; When setting the concentration of AuNPs-ACA-2 solution as 0, gained green intensity blank sample intensity is designated as F0, calculate the fluorescent quenching efficiency (F corresponding to variable concentrations AuNPs-ACA-2 solution0-F)/F0, obtain fluorescent quenching efficiency maximum time corresponding AuNPs-ACA-2 solution concentration C;
(4) standard solution of ***e is prepared: prepare the ***e solution of variable concentrations with body fluid, be ***e standard solution;
(5) ***e examination criteria curve is drawn: in the hydrophilic detection region of the paper substrate detecting device described in step (1), add volume be respectively the ***e standard solution of V1 and the AuNPs-ACA-2 solution of volume to be V2 concentration be C, after having reacted, remove unreacted AuNPs-ACA-2; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region and read green intensity F ' in fluoroscopic image with software;When setting the concentration of ***e standard solution as 0, gained green intensity blank sample intensity is designated as F0', with fluorescent quenching efficiency (F0’-F’)/F0' for vertical coordinate, with the logarithm value of ***e concentration of standard solution for abscissa drawing standard curve;
(6) mensuration of ***e concentration in body fluid to be measured: add volume respectively in the hydrophilic detection region of the paper substrate detecting device described in step (1) and be the body fluid to be measured of V1 and the AuNPs-ACA-2 solution of volume to be V2 concentration be C, remove unreacted AuNPs-ACA-2 after having reacted; Reacted for gained paper substrate detecting device is excited with 980nm LASER Light Source after drying, under dark condition, shoots the fluoroscopic image in hydrophilic detection region and read green intensity Fx in fluoroscopic image with software; Calculate fluorescent quenching efficiency (F0’-Fx)/F0', substitute into standard curve and obtain the ***e concentration in body fluid to be measured.
2. a kind of ***e fast quantitative measurement method for detecting suitable in Site Detection according to claim 1, it is characterised in that described hydrophilic detection region generally circular in shape, internal diameter is 2-6mm.
3. a kind of ***e fast quantitative measurement method for detecting suitable in Site Detection according to claim 1, it is characterised in that described water-soluble upconversion fluorescence nano material pattern is spherical, and particle diameter is at 30-100nm, and finishing has amino.
4. a kind of ***e fast quantitative measurement method for detecting suitable in Site Detection according to claim 1, it is characterised in that described gold nano grain pattern is spherical, and particle diameter is at 10-30nm, and finishing has citrate.
5. a kind of ***e fast quantitative measurement method for detecting suitable in Site Detection according to claim 1, it is characterised in that described single-chain nucleic acid ACA-1 and single-chain nucleic acid ACA-2, its sequence is respectively as follows: 5 '-NH2-TTTTTCAAGAACTGAGGG-3 ' and 5 '-SH-TTTTTACAGCAGGGTGAAGTAACTTCTTG-3 '.
6. a kind of ***e fast quantitative measurement method for detecting suitable in Site Detection according to claim 1, it is characterised in that the buffer solution described in step (2) is Tris-HCl buffer solution, and pH scope is at 7.0-7.5.
7. a kind of ***e fast quantitative measurement method for detecting suitable in Site Detection according to claim 1, it is characterised in that in step (3), the concentration range of AuNPs-ACA-2 solution is 0-0.305 μM.
8. a kind of ***e fast quantitative measurement method for detecting suitable in Site Detection according to claim 1, it is characterized in that in step (3) with buffer solution to remove unreacted AuNPs-ACA-2, the pH of described buffer ranges for 7.0-7.5.
9. a kind of ***e fast quantitative measurement method for detecting suitable in Site Detection according to claim 1, it is characterised in that the concentration of ***e standard solution respectively 0-50 μM in described step (4).
10. a kind of ***e fast quantitative measurement method for detecting suitable in Site Detection according to claim 1, it is characterized in that described step (3) and step (5), (6) adopting mobile phone shoot hydrophilic detection region fluoroscopic image, read green intensity in fluoroscopic image with mobile phone RGB software.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106940372A (en) * | 2017-03-06 | 2017-07-11 | 南京中医药大学 | Detect methylene blue, upper conversion nano grain modification fibrosis paper sensor of Telomerase and preparation method thereof |
| CN109116040A (en) * | 2018-08-21 | 2019-01-01 | 江苏大学 | A method of ***e is detected based on double sulfydryl aptamers |
| CN112763440A (en) * | 2020-12-29 | 2021-05-07 | 西安邮电大学 | Method for detecting thiram based on silver nano triangular plate |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040023216A1 (en) * | 2002-05-10 | 2004-02-05 | Board Of Trustees Of The University Of Illinois | Fluorescence based biosensor |
| CN101788489A (en) * | 2010-02-10 | 2010-07-28 | 中国政法大学 | Sensitive thin-film material used for detecting drugs efficiently and preparation method thereof |
| CN101948907A (en) * | 2009-10-30 | 2011-01-19 | 中国科学院上海微***与信息技术研究所 | Method for improving detection sensitivity of ***e |
| US20110171749A1 (en) * | 2009-03-02 | 2011-07-14 | Board Of Trustees Of Michigan State University | Nanoparticle tracer-based electrochemical dna sensor for detection of pathogens-amplification by a universal nano-tracer (aunt) |
| CN102650612A (en) * | 2011-02-25 | 2012-08-29 | 首都师范大学 | Method for ***e electrochemical detection and three-sectioned adapter probe used therein |
| CN102676508A (en) * | 2012-04-20 | 2012-09-19 | 华森新科（苏州）纳米技术有限公司 | Small molecule probe based on nano-gold and aptamer and preparation method of small molecule probe |
-
2016
- 2016-01-20 CN CN201610037309.7A patent/CN105675563B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040023216A1 (en) * | 2002-05-10 | 2004-02-05 | Board Of Trustees Of The University Of Illinois | Fluorescence based biosensor |
| US20110171749A1 (en) * | 2009-03-02 | 2011-07-14 | Board Of Trustees Of Michigan State University | Nanoparticle tracer-based electrochemical dna sensor for detection of pathogens-amplification by a universal nano-tracer (aunt) |
| CN101948907A (en) * | 2009-10-30 | 2011-01-19 | 中国科学院上海微***与信息技术研究所 | Method for improving detection sensitivity of ***e |
| CN101788489A (en) * | 2010-02-10 | 2010-07-28 | 中国政法大学 | Sensitive thin-film material used for detecting drugs efficiently and preparation method thereof |
| CN102650612A (en) * | 2011-02-25 | 2012-08-29 | 首都师范大学 | Method for ***e electrochemical detection and three-sectioned adapter probe used therein |
| CN102676508A (en) * | 2012-04-20 | 2012-09-19 | 华森新科（苏州）纳米技术有限公司 | Small molecule probe based on nano-gold and aptamer and preparation method of small molecule probe |
Non-Patent Citations (1)
| Title |
|---|
| MILAN N.STOJANOVIS .ET AL: "Aptamer-Based Folding Fluorescent Sensor for Cocaine", 《J.AM.CHEM.SOC》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106940372A (en) * | 2017-03-06 | 2017-07-11 | 南京中医药大学 | Detect methylene blue, upper conversion nano grain modification fibrosis paper sensor of Telomerase and preparation method thereof |
| CN109116040A (en) * | 2018-08-21 | 2019-01-01 | 江苏大学 | A method of ***e is detected based on double sulfydryl aptamers |
| CN112763440A (en) * | 2020-12-29 | 2021-05-07 | 西安邮电大学 | Method for detecting thiram based on silver nano triangular plate |
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|---|---|
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