CN107084951A - A kind of method that aptamers recognition detection zearalenone is marked based on time-resolved fluorescence - Google Patents
A kind of method that aptamers recognition detection zearalenone is marked based on time-resolved fluorescence Download PDFInfo
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- CN107084951A CN107084951A CN201710232945.XA CN201710232945A CN107084951A CN 107084951 A CN107084951 A CN 107084951A CN 201710232945 A CN201710232945 A CN 201710232945A CN 107084951 A CN107084951 A CN 107084951A
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- G—PHYSICS
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- 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
- G01N21/6408—Fluorescence; Phosphorescence with measurement of decay time, time resolved fluorescence
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- G—PHYSICS
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- 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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Abstract
The invention provides a kind of method that aptamers recognition detection zearalenone is marked based on time-resolved fluorescence, for wheat, cereal, the detection of zearalenone content in feed and its product etc..The characteristics of present invention makes full use of time-resolved fluorescence nano material long fluorescence lifetime, it is prevented effectively from the interference of sample biological context fluorescence, improve detection sensitivity, in combination with zearalenone specific nucleic acid aptamers functional magnetic nano material Magneto separate example enrichment effect, large sample can be concentrated quickly and effectively improves thousands of times of concentration of specimens, detection cycle can be greatly shortened.
Description
Technical field
It the present invention relates to the use of magnetic Nano material and the identification pair of time-resolved fluorescence nano material combination aptamer
Zearalenone is detected that feature is the magnetic Nano material using zearalenone aptamers functionalization to corn
Zearalenone separation and concentration in the samples such as wheat so that easy to detect quick, while utilizing time-resolved fluorescence nanometer material
Material further improves the sensitivity of detection as label by detection time resolved fluorometric signal, and this method belongs to nano material
With field of molecular biotechnology.
Background technology
Zearalenone (Zearalenone, ZEN), also known as F-2 toxin are main by sickle-like bacteria such as Fusarium graminearums
A kind of non steroidal estrogen sample effect mycotoxin produced.The chemical name of the toxin is 6- (10- hydroxyl -6- epoxides-ten one
Carbene base)-β-thunder lock acid, molecular formula：C18H22O5, relative molecular mass 318.4 is insoluble in water, be soluble in alkaline aqueous solution and
Alcohols, its methanol solution is in green-blue fluorescence under ultraviolet light.ZEN is widely present in cereal, mainly pollutes corn, barley, small
The crops such as wheat, oat, sorghum and rice, also have to a certain degree in the product such as flour, malt, beer and dairy produce, meat products
Distribution.ZEN causes serious threat to the health of the mankind and animal, and harm is only second to aflatoxin.ZEN to animal have compared with
Strong Reproductive and developmental toxicity and immunotoxicity, can cause the hyperfunction disease of animal estrogen, cause infertile, fetal anomaly and growth
Depauperation, the especially influence to pig, ox and sheep are larger, and huge economic loss is brought to animal husbandry.When human consumption is dirty
After the product such as the crop of dye or meat, egg milk, ZEN shows as Reproductive and developmental toxicity, immunotoxicity, gene into human body main harm
Toxicity, liver renal toxicity, cause tumour (hysteroma, breast cancer, carcinoma of testis) toxicity.
At present, ZEN detection methods mainly have thin-layered chromatography (TLC), high performance liquid chromatography (HPLC), efficient liquid phase matter
Compose combination method (HPLC-MS/MS) and the immunization set up based on Ag-Ab Immune discrimination etc..TLC operating procedure
Cumbersome, sensitivity and specificity are low, the need for can not meeting modern measure.And locate before conventional red, orange, green, blue, yellow (ROGBY), sample
Reason method is complicated, cumbersome, and needs skilled grasp instrumentation technology, and inconvenient basic unit promotes.Though immunization detection ZEN has spirit
The advantages of sensitivity height, high specificity, but small molecule antigens ZEN preparation process is cumbersome time-consuming, costly, between different batches
Having differences property of antibody, storage and use condition are harsh, cause false positive rate height, poor repeatability.So a kind of quick side of exploitation
Just, stability is good, sensitivity is high, high specificity, detection method with low cost are necessary.
Aptamer (Aptamer) is by index concentration part from the random oligonucleotide library synthesized in vitro
Phyletic evolution (Systematic Evolution of Ligands by Exponential Enrichment, SELEX) technology
Screen the obtained cluster small molecule DNA specifically bound with target substance or RNA fragments.Aptamer can recognize right therewith
The target substances such as any kind of albumen and low molecule answered, and there is high affinity with target substance.Compare, adjust suitable with antibody
Part has many advantages, such as that such as external synthesis cycle is short, and preparation cost is low, without zoopery；Stability is good, unwise to temperature
Sense；Be easy to modification etc., and aptamers as identification molecule in clinical diagnosis, clinical treatment, medicament transport, protein
It is used widely in group research and food security.
Time-resolved fluorescence is a kind of special fluorescence phenomenon, is to carry out the time point using the fluorescence lifetime length of material
Distinguish, i.e., using appropriate excitation source and detection architecture, can obtain in fluorescence intensity-time graph of fixed wave length and solid
The fluorescence emission spectrum fixed time, realizes for the component to spectra overlapping in mixture but aging variation and differentiates, and can distinguish
It is measured；By setting rational time delay (Delay time) and gate duration (Gate time, also referred to as detection time)
The interference of impurity autofluorescence and background fluorescence to signal to noise ratio in detection environment is eliminated, so as to improve sensitivity of analytical method.
The key of Time-resolved fluorescence assay technology development is effective exploitation and the combination of lanthanide series fluorescence probe and analytical model
Utilize.With flourishing for nanometer technology, the group of the lanthanides nano material with superior water dispersibility is used as a class novel light-emitting mark
Remember thing, its intrinsic time resolution optical characteristics is gradually developed and used.Lanthanide doped nano material has all in itself
More superior physicochemical property, such as water dispersible are good, fluorescence lifetime length, resistance to photobleaching, Stokes shift are wide, bio-compatibility
Good, cytotoxicity is low, polychrome controllable (doped chemical species and ratio), is conducive to multicomponent in biosystem to detect simultaneously.
In view of above-mentioned advantage, new lanthanide doped fluorescent nano material possesses good Time-resolved fluorescence assay application prospect.
The physical length of magnetic Nano material just at nanometer scale and with preparation method is simple, raw material is cheap, it is unique super suitable
The features such as magnetic, surface are easy to modification, good biocompatibility so that it has broad application prospects in biomedical sector.
Present invention synthesis first has obtained the magnetic Nano material and time-resolved fluorescence nanometer material of surface biological functionalization
Material, then the nucleic acid adaptation that the target nucleic acid aptamers and time-resolved fluorescence nano material combined by magnetic Nano material are combined
Both are assembled composition composite nanostructure by the hybridization between body complementary strand, are excited with ultraviolet light, and induction time is differentiated glimmering
Light is detection signal, by the detection to various concentrations zearalenone standard items, sets up standard curve, reaches to containing corn
The purpose that zeranol sample is quantitatively detected.The invention can be used for the samples such as corn, wheat, cereal, feed and its product
The detection of zearalenone content in product.
The content of the invention
A kind of method of time-resolved fluorescence mark-aptamers recognition detection zearalenone：Ammonia is prepared respectively
The Fe of base3O4Magnetic Nano material and time-resolved fluorescence nano material NaYF4:Ce/Tb, to magnetic Nano material and time
Fluorescence differentiates nano material and carries out surface Avidin (Avidin) modification, then passes through Avidin (Avidin) and biotin
(Biotin) the aptamers DNA and aptamers complementary strand (cDNA) specificity that the effect between is modified with biotin (Biotin) are tied
Conjunction respectively constitutes fluorescence probe and capture probe.By both through incubation after a while, by Complementary hybridization so that both groups
Dress up composite nano materials.Nano material is separated using external magnetic field and excited using ultraviolet light (273nm), is recorded
Now fluorescence signal intensity is maximum, when adding target analytes zearalenone, and the space conformation of aptamers occurs
Change, specifically binds so that aptamers DNA unwinds with complementary single strand (cDNA), so as to cause with zearalenone
Time-resolved fluorescence nano material is separated with magnetic nanoparticle, is detected after being now enriched with again, obtains fluorescence signal intensity reduction.
The content of zearalenone is related to the numerical value that fluorescence signal intensity reduces within the specific limits, based on this to Gibberella zeae alkene
Ketone standard items are detected, set up standard curve, to reach the mesh quantitatively detected to zearalenone in actual sample
's；Step is：
1st, using a step solvent structure NaYF4:Ce/Tb simultaneously does surface modification to it
The NaCl of the phosphoethanolamine (AEP) and 1mmol that weigh 1mmol is dissolved in 30mL ethylene glycol and is stirred continuously, with
0.9mmol Y (NO are added afterwards3)3·6H2O、0.05mmol Ce(NO3)3·6H2O、0.05mmol Tb(NO3)3·6H2O.So
Afterwards, 4mmol NH will be contained4Above-mentioned clear solution is added dropwise in F 10mL ethylene glycol solutions (45 DEG C of stirring in water bath fully dissolve)
In, continue to be transferred in 50mL Teflon autoclaves after stirring 30min at room temperature, 195 DEG C of reaction 4h.After reaction, take
Go out to be cooled to room temperature, with alternately washing three times of ethanol, ultra-pure water, be dissolved in ultra-pure water and being saved backup in 4 DEG C of refrigerators.
2nd, amination Fe is prepared3O4Magnetic nanoparticle
Weigh 2.0g anhydrous sodium acetates and 1.0g Iron(III) chloride hexahydrates add 30mL ethylene glycol, by the 1,6- of 6.5g warm bath
Hexamethylene diamine is added in the former mixed liquor, is heated to 50 DEG C, and is stirred continuously to form the bright homogeneous colloidal solution of claret.Will
The solution is transferred in 100mL reactors, 198 DEG C of pyroreaction 6h.Reactor is taken out after the completion of reaction and naturally cools to room temperature
Beaker (rinsing transfer residual powder in batches) is all poured into, is then separated and collected using magnet, discards upper strata mixing liquid, by magnetic
Separate the black powder ultra-pure water obtained, ultrasonic disperse, then Magneto separate discard solution, collect powder.Using ultra-pure water and
Absolute ethyl alcohol is washed three times in turn, and gained black powder dries 12h at 50 DEG C, is taken out mortar grinder to fine powder, is stored standby
With.
3rd, using glutaraldehyde method by time-resolved fluorescence nano material and magnetic Nano material and Avidin (Avidin)
It is coupled
Because amino group is contained on time-resolved fluorescence nano material and magnetic Nano material surface, therefore penta can be passed through
Avidin is covalently bound to material surface by dialdehyde method.Time fluorescence is differentiated into nano material and magnetic Nano material 10mM phosphorus
Phthalate buffer (PBS) is resuspended, and 1mg/mL solution is made, ultrasonic 15min makes it be well-dispersed in buffer solution.By material with
Lucifuge slightly shakes activation 2h to 5% glutaraldehyde at room temperature, and the method for time-resolved fluorescence nano material centrifugation is removed not
The glutaraldehyde of reaction, magnetic material removes unreacted glutaraldehyde by externally-applied magnetic field, then with PBS three times, Ran Houjia
Enter 37 DEG C of concussions of Avidin (1mg/mL) to stay overnight.Reaction is cleaned for several times after terminating, and abandons supernatant.
4th, Avidin modification magnetic Nano material and time-resolved fluorescence nano material respectively with aptamers and aptamers
Complementary strand is coupled
Using by the specific binding between Avidin (Avidin) and biotin (Biotin) that surface modification is affine
The Gibberella zeae that the magnetic Nano material and time-resolved fluorescence nano material of plain (Avidin) are modified with biotin (Biotin)
Ketenes aptamers DNA is single-stranded and the cDNA connections of aptamers complementary strand.Specific method is：The magnetic Nano that 5mg Avidins are modified
Material and time-resolved fluorescence nano material are scattered in 5mL (10mM) phosphate buffer, are separately added into a certain amount of biology
The zearalenone aptamers and aptamers complementary strand of elementization, are incubated 12h in 37 DEG C of shaking tables, material are collected by centrifugation, use phosphorus
Phthalate buffer is cleaned three times, is scattered in phosphate buffer.
5th, zearalenone aptamers DNA is single-stranded and aptamers complementary strand cDNA hybridizes
Time-resolved fluorescence nano particle and magnetic Nano material are connected into one nano-complex of composition, it is specific real
Applying method：The single-stranded solution of complementary DNA for taking 400 μ L time-resolved fluorescences nano materials to mark, with 100 μ L aptamers functionalization magnetic
Property nano material mixing, in phosphoric acid buffer liquid system 37 DEG C reaction 1h, afterwards in additional magnetic fields 1min, collection is assembled
Nano material compound, and cleaned 3 times with phosphate buffer.
6th, zearalenone standard items are detected, set up standard curve
In the phosphate buffer that the nano material compound assembled is dissolved in, the Gibberella zeae of various concentrations is prepared
Ketenes is added in complex systems, and 40min is incubated under the conditions of 37 DEG C, then abandons supernatant by Magneto separate, cleans three times to ensure
The time-resolved fluorescence nano material that comes off is dissociated not in magnetic Nano material surface attachment, thus ensure detection sensitivity and
The mixture being enriched to, is finally resuspended in 200 μ L phosphate buffer and carries out upper machine testing by accuracy.Using ELIASA
Synergy H1, BioTek set excitation wavelength as 273nm under time-resolved fluorescence pattern, and time delay is 100 μ s, inspection
The survey time is 1000 μ s, has time-resolved fluorescence signal at 544nm, for detecting zearalenone.With Gibberella zeae
The increase of ketenes concentration, time-resolved fluorescence signal progressively reduces.According to fluorescent value and corresponding zearalenone standard items
Concentration sets up standard curve, and experimental result obtains good linear relationship in 0.001~10ng/mL intervals.
7th, zearalenone sample is detected
Simple processing is done to sample, is then added directly into above-mentioned nano composite system, is incubated under the conditions of 37 DEG C
40min, using under ELIASA time-resolved fluorescence pattern, excites the fluorescence signal obtained at 544nm under 273nm, bent from standard
The concentration of corresponding zearalenone is tried to achieve in line.
Advantages of the present invention：
1st, using lanthanide doped nano material fluorescence lifetime it is long the characteristics of, passage time differentiate cause detection background fluorescence
Decay, so as to greatly improve the sensitivity of detection.
2nd, the specific binding using aptamers to detection material, effectively raises the Stability and veracity of detection.
3rd, by the use of magnetic Nano material as capture probe, target detection thing is enriched with, can effectively shorten detection
It is cycle, simple to operate.
Brief description of the drawings
Fig. 1 is the schematic diagram based on time-resolved fluorescence mark-aptamers recognition detection zearalenone
Fig. 2 is time-resolved fluorescence intensity with zearalenone change in concentration stacking chart (a)；Zearalenone is detected
Canonical plotting (b).
Embodiment：
The present invention includes but is not limited to following examples, it is all carried out under the spirit and principles in the present invention any equally replace
Change or refuse to improve, all will be regarded as within protection scope of the present invention.
Embodiment 1：The detection of zearalenone and mark-on reclaims in corn actual sample
Sample pretreatment：Corn pulverizes and sieves at a high speed, weighs 5g in 100mL flasks, adds 5g NaCl, methanol (7:3)
The aqueous solution, is placed in the stirring of homogenizer high speed and extracts 2min, static a moment, filtering takes 10mL filtrates to be placed in after fully mixing
In 50mL flasks, extract 2min in the stirring of homogenizer high speed again, it is static after filtered with ultra-fine fibre glass filter paper, until
Filtrate is clarified, and collects filtrate standby.Respectively choose 0.001,0.01,0.1,0.5,1,5, the Gibberella zeae alkene of 10ng/mL concentration
Ketone standard items are added in determinand, detect the content of wherein zearalenone again also with the inventive method, obtain
Detected value.Rate of recovery %=(detected value-background values)/addition × 100%.From the point of view of the result of table one, the rate of recovery is 80.76
~122.36%, illustrate that the present invention is stable, it is sensitive, accurately, it is suitable for the detection of zearalenone in corn actual sample.
Table one：The detection of zearalenone and recovery of standard addition in corn actual sample
Embodiment 2：The detection of zearalenone and mark-on reclaims in wheat actual sample
Sample pretreatment is with example 1.Choose 0.001 respectively, 0.01,0.1,0.5,1,5, the corn of 10ng/mL concentration it is red
Mould ketenes standard items are added in determinand, detect the content of wherein zearalenone again also with the inventive method,
Obtain detected value.Rate of recovery %=(detected value-background values)/addition × 100%.From the point of view of the result of table two, the rate of recovery exists
90.04~114.75%, illustrate that the present invention is equally suitable for the detection of zearalenone in wheat actual sample.
Table two：The detection of zearalenone and recovery of standard addition in wheat actual sample
Claims (3)
1. a kind of method based on time-resolved fluorescence mark-aptamers recognition detection zearalenone, it is characterised in that warp
Cross assembling and obtain zearalenone aptamers functional magnetic nano material and surface modification aptamers complementary single strands
Time-resolved fluorescence nano material, then both are assembled by composite nanostructure by DNA Complementary hybridizations, obtained with ultraviolet excitation
Time-resolved fluorescence detects signal.When adding target analytes zearalenone, the space conformation of aptamers changes
Specifically bound with zearalenone so that aptamers are unwind with aptamers complementary single strand, so as to cause part-time
Resolved fluorometric nano material is separated with magnetic Nano material, the time-resolved fluorescence nano material of magnetic Nano Nanosurface assembling
Fluorescence signal intensity reduce, zearalenone standard items are detected based on this, standard curve is set up, with reach to containing
The purpose that zearalenone sample is detected.
2. it is as claimed in claim 1 a kind of based on time-resolved fluorescence mark-aptamers recognition detection zearalenone
Method, it is characterised in that first to amidized magnetic Nano material surface modification Avidin (Avidin), pass through Avidin afterwards
(Avidin) effect between biotin (Biotin) is specifically bound with the aptamers DNA that biotin (Biotin) is modified,
As Magneto separate reagent.
3. it is as claimed in claim 1 a kind of based on time-resolved fluorescence mark-aptamers recognition detection zearalenone
Method, it is characterised in that what the time-resolved fluorescence nano material of Avidin (Avidin) modification was modified with biotin (Biotin)
The single-stranded combination of aptamer complementary nucleic acid, constitutes nucleic acid marking probe.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107515295A (en) * | 2017-10-17 | 2017-12-26 | 环境保护部华南环境科学研究所 | A kind of method of mycotoxin in detection environment |
| CN110330974A (en) * | 2019-07-11 | 2019-10-15 | 南京工业大学 | A kind of preparation and application of zearalenone ratio fluorescent probe |
| CN112255214A (en) * | 2020-10-21 | 2021-01-22 | 江南大学 | Method for simultaneously detecting three mycotoxins based on time-resolved fluorescence labeling-aptamer recognition |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102023147A (en) * | 2010-09-29 | 2011-04-20 | 江南大学 | Method for detecting ochratoxin A by magnetic separation of aptamer-functionalized magnetic nano material and marking of up-conversion fluorescent nano material |
| CN102443585A (en) * | 2011-11-25 | 2012-05-09 | 国家纳米技术与工程研究院 | Zearalenone nucleic acid aptamer and application thereof |
| CN103013998A (en) * | 2012-11-22 | 2013-04-03 | 江南大学 | Oligonucleotides aptamer special for distinguishing zearalenone |
| US20130171623A1 (en) * | 2012-01-02 | 2013-07-04 | Aimin He | Binding Assays Utilizing Time-Resolved Up-Converting Luminescence Detection |
| CN103352077A (en) * | 2011-09-08 | 2013-10-16 | 苏州友林生物科技有限公司 | Mammaglobin mRNA detection method and reagent thereof |
-
2017
- 2017-04-11 CN CN201710232945.XA patent/CN107084951B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102023147A (en) * | 2010-09-29 | 2011-04-20 | 江南大学 | Method for detecting ochratoxin A by magnetic separation of aptamer-functionalized magnetic nano material and marking of up-conversion fluorescent nano material |
| CN103352077A (en) * | 2011-09-08 | 2013-10-16 | 苏州友林生物科技有限公司 | Mammaglobin mRNA detection method and reagent thereof |
| CN102443585A (en) * | 2011-11-25 | 2012-05-09 | 国家纳米技术与工程研究院 | Zearalenone nucleic acid aptamer and application thereof |
| US20130171623A1 (en) * | 2012-01-02 | 2013-07-04 | Aimin He | Binding Assays Utilizing Time-Resolved Up-Converting Luminescence Detection |
| CN103013998A (en) * | 2012-11-22 | 2013-04-03 | 江南大学 | Oligonucleotides aptamer special for distinguishing zearalenone |
Non-Patent Citations (3)
| Title |
|---|
| DATAO TU ET AL: "Time-Resolved FRET Biosensor Based on Amine-Functionalized Lanthanide-Doped NaYF4 Nanocrystals", 《ANGEW. CHEM. INT. ED》 * |
| XIAOLE WANG ET AL: "Simultaneous detection of Staphylococcus aureus and Salmonella typhimurium using multicolor time-resolved fluorescence nanoparticles as labels", 《INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY》 * |
| 马智鸿等: "应用生物素-链霉亲和素的时间分辨荧光免疫分析检测玉米赤霉烯酮", 《江苏农业学报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107515295A (en) * | 2017-10-17 | 2017-12-26 | 环境保护部华南环境科学研究所 | A kind of method of mycotoxin in detection environment |
| CN110330974A (en) * | 2019-07-11 | 2019-10-15 | 南京工业大学 | A kind of preparation and application of zearalenone ratio fluorescent probe |
| CN110330974B (en) * | 2019-07-11 | 2022-09-09 | 南京工业大学 | Preparation and application of zearalenone ratiometric fluorescent probe |
| CN112255214A (en) * | 2020-10-21 | 2021-01-22 | 江南大学 | Method for simultaneously detecting three mycotoxins based on time-resolved fluorescence labeling-aptamer recognition |
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