CN103808701A - Homogeneous-phase rapid detection method of ochratoxin A based on nucleic acid chimeric dye fluorescence quenching - Google Patents
Homogeneous-phase rapid detection method of ochratoxin A based on nucleic acid chimeric dye fluorescence quenching Download PDFInfo
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- CN103808701A CN103808701A CN201310429041.8A CN201310429041A CN103808701A CN 103808701 A CN103808701 A CN 103808701A CN 201310429041 A CN201310429041 A CN 201310429041A CN 103808701 A CN103808701 A CN 103808701A
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Abstract
The invention discloses a homogeneous-phase rapid detection method of ochratoxin A based on nucleic acid chimeric dye fluorescence quenching. The homogeneous-phase rapid detection method comprises the following steps: (1) forming an ochratoxin A nucleic acid aptamer and a nucleic acid chimeric dye fluorescence composite; and (2) specifically recognizing ochratoxin A by the ochratoxin A nucleic acid aptamer. According to the invention, the OTA (ochratoxin A) detection method is constructed by taking the ochratoxin A nucleic acid aptamer as a molecule recognition element and taking the nucleic acid chimeric dye as report molecule; the cost for detecting OTA fungaltoxin is reduced greatly; the detection method is good in reproducibility and high in specificity, is simple and convenient to operate and can detect a great quantity of samples in parallel. The homogeneous-phase rapid detection method provided by the invention can detect OTA only for 5-10 minutes after the ochratoxin A nucleic acid aptamer and the nucleic acid chimeric dye fluorescence composite are formed in advance.
Description
Technical field
The present invention relates to utilize the fluorescent quenching of the chimeric dyestuff of nucleic acid to detect ochratoxin A, feature is the application of ochratoxin A aptamer and the chimeric dyestuff of nucleic acid, and the detection speed of the method is fast, detection flux is high and strong operability, belongs to detection technique of fluorescence field.
Background technology
Ochratoxin A (OchratoxinA, OTA) mainly by aspergillus He Celadon mould wait generation, be widespread in nature, wherein with in the majority on the crops such as grain (wheat, corn, barley, oat, rye, rice and broomcorn millet class etc.), peanut, vegetables (beans), there are many Poisonings such as liver renal toxicity and very strong teratogenesis, carcinogenic and mutagenesis, animal and human's body health is had to very large potential hazard.Given this, detection and the control to OTA is all paid attention in countries in the world, has made one after another grain limit standard relevant with OTA in feed, as the maximum limitation of the regulation cereal materials OTA of European Union 5 μ g/kg, the maximum limitation of cereal fabricated product 3 μ g/kg; The maximum limitation of OTA 5 μ g/kg in China's regulation cereal, beans, to protect people ' s health and regulation and control to import and export grain.
The main method that detects at present OTA is thin-layered chromatography (TLC), high performance liquid chromatography (HPLC), enzyme linked immunosorbent assay (ELISA) etc.TLC method is simple, and testing cost is low, but sensitivity is poor, reappearance is bad and cannot realize robotization; HPLC method is highly sensitive, but required instrument costliness, complex pretreatment, testing cost are high, is not suitable for the rapid screening of a large amount of samples.ELISA method is highly sensitive, specificity good, but the manufacturing cycle of the high-quality antibody relying on is very long and have a problem of cross reaction.
Aptamer is as a class single stranded oligonucleotide, before and after interacting with target molecule generation specificity can there is corresponding variation in space conformation, having compared with antibody can in-vitro screening acquisition, Heat stability is good, be easy to the advantage such as chemosynthesis and modification, even can distinguish the target molecule of the single substituting group difference that monoclonal antibody cannot realize; The chimeric dyestuff of nucleic acid has following features: under free state without fluorescence or there is very weak fluorescence, be fitted to and fluorescence occurs in DNA double chain strengthen, therefore the homogeneous fluorescent detection method based on the chimeric dyestuff dyestuff of nucleic acid has highly sensitive, simple to operate and can carry out the feature such as Parallel testing of a large amount of samples.So the homogeneous fluorescent detection method that the high specific of aptamer, high-affinity are combined with the chimeric dyestuff of nucleic acid, can overcome the shortcoming of the main detection method of current OTA well, is expected to become a kind of potential quick screening method.
Summary of the invention
In order to solve, scheme complexity, the operation of current OTA detection method locked, high cost, detection time long shortcoming, the invention provides a kind of ochratoxin A homogeneous phase method for quick based on the chimeric dye fluorescence cancellation of nucleic acid.
Technical scheme of the present invention is that the ochratoxin A homogeneous phase method for quick based on the chimeric dye fluorescence cancellation of nucleic acid, comprises the following steps:
(1) formation of the chimeric dye fluorescence compound of ochratoxin A aptamer and nucleic acid;
(2) specific recognition of ochratoxin A aptamer to ochratoxin A.
The concrete operation step that the present invention is based on the ochratoxin A homogeneous phase method for quick that the chimeric dye fluorescence cancellation of nucleic acid realizes is as follows:
The formation of the chimeric dye fluorescence compound of ochratoxin A aptamer and nucleic acid
First ochratoxin A aptamer is carried out to pre-service: 95 ℃ of sex change 5min, 4 ℃ of cooling 5min.Then get 50 μ L10mM Tris-HCl reaction buffers (pH8.5), wherein the concentration of ochratoxin A aptamer is 50nM, the dilution ratio of the chimeric dyestuff of GeneFinder nucleic acid is 8:10000, room temperature reaction 30min, so that the chimeric dyestuff of nucleic acid and the abundant combination of ochratoxin A aptamer form ochratoxin A aptamer and the chimeric dye fluorescence compound of nucleic acid; Ochratoxin A aptamer is ssDNA probe, and sequence is 5 '-CTTCGGGTGTGGGTGGCATTCCGGTAGGCTCTG-3 '.
The specific recognition of ochratoxin A aptamer to ochratoxin A
In the reacted solution of above-mentioned steps (1), add appropriate ochratoxin A solution, adjusting liquor capacity with 10mM Tris-HCl reaction buffer (pH8.5) is 100 μ L, wherein the concentration of ochratoxin A aptamer is 25nM, the dilution ratio of the chimeric dyestuff of GeneFinder nucleic acid is 4:10000, room temperature reaction 5-10min, so that the abundant combination of ochratoxin A aptamer and ochratoxin A, form ochratoxin A aptamer and ochratoxin A compound, thereby cause dissociating of the chimeric dye fluorescence compound of ochratoxin A aptamer and nucleic acid, produce the variation of fluorescence intensity.Reacted solution is transferred in micro-fluorescence cuvette and puts into fluorescence spectrophotometer, or reacted solution is shifted in microwell plate and puts into the fluorescence spectrophotometer with microwell plate read module, take 490nm as excitation wavelength, 525nm is emission wavelength, the variation of measuring its fluorescence signal intensity and recording fluorescence intensity: the fluorescence intensity (I of blank
0) maximum, along with the increase fluorescence intensity (I) of OTA concentration weakens gradually.
By above-mentioned (1), (2) step, the variable concentrations ochratoxin A titer of preparation is detected, record changing value (the Δ I=I of the fluorescence intensity that each standard solution sample produces
0-I).With Δ I, to ochratoxin A concentration mapping acquisition typical curve in standard solution, the changing value of the fluorescence intensity that in unknown sample, ochratoxin A produces contrasts and determines its concentration with mark curve.
Beneficial effect imbody of the present invention is in the following areas in sum:
1) the present invention adopts ochratoxin A aptamer as molecular recognition elements, the OTA detection method that the chimeric dyestuff of nucleic acid builds as reporter molecules, greatly reduce the cost that detects OTA mycotoxin, favorable reproducibility, specificity is high, simple to operation and can realize the Parallel testing of a large amount of samples;
2) the present invention is based on the ochratoxin A homogeneous phase method for quick of the chimeric dye fluorescence cancellation of nucleic acid, after ochratoxin A aptamer and the chimeric dye fluorescence compound of nucleic acid are pre-formed, only need 5-10min can realize the detection to OTA.
Accompanying drawing explanation
Fig. 1: the phenomenon figure of the ochratoxin A homogeneous phase method for quick based on the chimeric dye fluorescence cancellation of nucleic acid: wherein (a) is GeneFinder+Aptamer+AFB1, (b) be GeneFinder+Aptamer, (c) be GeneFinder+Aptamer+OTA, (d) being GeneFinder, is (e) OTA aptamer.[OTA?aptamer]=25nM,[OTA]=[AFB1]=50nM，Dilution?ratio?of?GeneFinder=4:10000。AFB1 is aflatoxin B1.
Fig. 2: the working curve diagram that ochratoxin A standard solution is detected.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.
(1) formation of the chimeric dye fluorescence compound of ochratoxin A aptamer and nucleic acid
First ochratoxin A aptamer is carried out to pre-service: 95 ℃ of sex change 5min, 4 ℃ of cooling 5min.Then get 50 μ L10mM Tris-HCl reaction buffers (pH8.5), wherein the concentration of ochratoxin A aptamer is 50nM, the dilution ratio of the chimeric dyestuff of GeneFinder nucleic acid is 8:10000, room temperature reaction 30min, so that the chimeric dyestuff of nucleic acid and the abundant combination of ochratoxin A aptamer form ochratoxin A aptamer and the chimeric dye fluorescence compound of nucleic acid; Ochratoxin A aptamer is ssDNA probe, and sequence is 5 '-CTTCGGGTGTGGGTGGCATTCCGGTAGGCTCTG-3 '.
A kind of screening preparation of ochratoxin A aptamer: ochratoxin A molecule is coupled on the magnetic bead of carboxyl modified by coupling agent EDC and NHS, with the buffer solution reaction of reacting that contains initial library, initial library is fixed on magnetic bead.Respectively take the magnetic bead of carboxyl modified as anti-Screening target; make (warfarin) and ochratoxin B molecule as negative Screening target take N-acetyl group-L-Phe (N-acetyl-L-phenylalanine), method China, screen for just screening molecule with ochratoxin A.In the situation that ochratoxin A exists, there is the oligonucleotide sequence of binding ability to disintegrate down from magnetic bead with it, after pcr amplification, adopt streptavidin magnesphere partition method to prepare secondary library, repeating to screen 15 takes turns, then carry out cloning and sequencing, finally carry out sequential analysis, active detection and sequence brachymemma and optimization, thereby obtain the specific nucleic acid aptamers of ochratoxin A.
(2) specific recognition of ochratoxin A aptamer to ochratoxin A
In the reacted solution of above-mentioned steps (1), add appropriate ochratoxin A solution, adjusting liquor capacity with 10mM Tris-HCl reaction buffer (pH8.5) is 100 μ L, wherein the concentration of ochratoxin A aptamer is 25nM, the dilution ratio of the chimeric dyestuff of GeneFinder nucleic acid is 4:10000, room temperature reaction 5-10min, so that the abundant combination of ochratoxin A aptamer and ochratoxin A, form ochratoxin A aptamer and ochratoxin A compound, thereby cause dissociating of the chimeric dye fluorescence compound of ochratoxin A aptamer and nucleic acid, produce the variation of fluorescence intensity.Reacted solution is transferred in micro-fluorescence cuvette and puts into fluorescence spectrophotometer, or reacted solution is shifted in microwell plate and puts into the fluorescence spectrophotometer with microwell plate read module, take 490nm as excitation wavelength, 525nm is emission wavelength, excite slit width to be set as 10nm, transmitting slit width is set as 5nm, the variation of measuring its fluorescence signal intensity and recording fluorescence intensity: the fluorescence intensity (I of blank
0) maximum, along with the increase fluorescence intensity (I) of OTA concentration weakens gradually.
As shown in Figure 1, the chimeric dyestuff of nucleic acid (GeneFinder) autofluorescence is very weak, negligible; OTA aptamer (aptamer) self does not have fluorescence; In the two common situation about existing, form aptamer/GeneFinder fluorescent composition, have very strong fluorescence, fluorescence strengthens more than 10 times.In the situation that OTA exists, the abundant combination of aptamer and OTA, forms aptamer/OTA compound, thereby causes dissociating of aptamer/GeneFinder fluorescent composition, produces fluorescent quenching, causes fluorescence intensity to reduce; And in the situation that aflatoxin B1 (AFB1) exists, aptamer nonrecognition AFB1, therefore aptamer/GeneFinder fluorescent composition can not dissociate, so can not produce fluorescent quenching, fluorescence intensity can not reduce.
By above-mentioned (1), variable concentrations ochratoxin A titer (0nM, 1nM, 2.5nM, 5.0nM, the 7.5nM of (2) step to preparation, 10nM, 15nM, 20nM, 25nM, 35nM, 50nM, 75nM) detect, and record changing value (the Δ I=I of the fluorescence intensity that each standard solution sample produces
0-I).With Δ I, to ochratoxin A concentration mapping acquisition working curve (Fig. 2) in standard solution, the changing value of the fluorescence intensity that in unknown sample, ochratoxin A produces contrasts with working curve determines its concentration.
Should be understood that, for those of ordinary skills, can be improved according to the above description or convert, and all these improvement and conversion all should belong to the protection domain of claims of the present invention.
SEQUENCE?LISTING
<110> applicant Henan Academy of Agricultural Sciences
The ochratoxin A homogeneous phase method for quick of <120> based on the chimeric dye fluorescence cancellation of nucleic acid
<130>
<160>1
<170>PatentIn?version3.5
<210>1
<211>33
<212>DNA
<213> ochratoxin A aptamer
<400>1
cttcgggtgt?gggtggcatt?ccggtaggct?ctg
33
Claims (3)
1. the ochratoxin A homogeneous phase method for quick based on the chimeric dye fluorescence cancellation of nucleic acid, its
Be characterised in that, comprise the following steps:
(1) formation of the chimeric dye fluorescence compound of ochratoxin A aptamer and nucleic acid;
(2) specific recognition of ochratoxin A aptamer to ochratoxin A.
2. method according to claim 1, is characterized in that, the concrete operation step of described method is as follows:
(1) formation of the chimeric dye fluorescence compound of ochratoxin A aptamer and nucleic acid
First ochratoxin A aptamer is carried out to pre-service: 95 ℃ of sex change 5min, 4 ℃ of cooling 5min.Then get 50 μ L10mM Tris-HCl reaction buffer solution (pH8.5), wherein the concentration of ochratoxin A aptamer is 50nM, the dilution ratio of the chimeric dyestuff of GeneFinder nucleic acid is 8:10000, room temperature reaction 30min, so that the chimeric dyestuff of nucleic acid and the abundant combination of ochratoxin A aptamer form ochratoxin A aptamer and the chimeric dye fluorescence compound of nucleic acid;
(2) specific recognition of ochratoxin A aptamer to ochratoxin A
In the reacted solution of above-mentioned steps (1), add appropriate ochratoxin A solution, adjusting liquor capacity with 10mM Tris-HCl reaction buffer (pH8.5) is 100 μ L, wherein the concentration of ochratoxin A aptamer is 25nM, the dilution ratio of the chimeric dyestuff of GeneFinder nucleic acid is 4:10000, room temperature reaction 5-10min, so that the abundant combination of ochratoxin A aptamer and ochratoxin A, form ochratoxin A aptamer and ochratoxin A compound, thereby cause dissociating of the chimeric dye fluorescence compound of ochratoxin A aptamer and nucleic acid, produce the variation of fluorescence intensity.Reacted solution is put into fluorescence spectrophotometer, take 490nm as excitation wavelength, 525nm is emission wavelength, the variation of measuring its fluorescence signal intensity and recording fluorescence intensity: the fluorescence intensity (I of blank
0) maximum, along with the increase fluorescence intensity (I) of OTA concentration weakens gradually.
By above-mentioned (1), (2) step, the variable concentrations ochratoxin A titer of preparation is detected, record changing value (the Δ I=I of the fluorescence intensity that each standard solution sample produces
0-I); With Δ I, to ochratoxin A concentration mapping acquisition typical curve in standard solution, the changing value of the fluorescence intensity that in unknown sample, ochratoxin A produces contrasts and determines its concentration with mark curve.
3. method according to claim 1, is characterized in that: ochratoxin A aptamer is ssDNA probe, and sequence is 5 '-CTTCGGGTGTGGGTGGCATTCCGGTAGGCTCTG-3 '.
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104178568A (en) * | 2014-07-25 | 2014-12-03 | 清华大学 | Method for detecting target substance in to-be-detected sample based on fluorescent sensing analysis of aptamer probe |
| CN104818279A (en) * | 2015-04-23 | 2015-08-05 | 河南省农业科学院 | Ochratoxin A nucleic acid adapter and application thereof to sample magnetic separation preprocessing |
| CN106124781A (en) * | 2016-07-07 | 2016-11-16 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of method based on aptamers sandwich fluorescent quenching technology for detection demethyl testosterone |
| CN106546724A (en) * | 2015-09-23 | 2017-03-29 | 中国医学科学院药用植物研究所 | A kind of new method of molecular beacon probe quick detection ochratoxin A |
| CN106706737A (en) * | 2016-12-10 | 2017-05-24 | 武汉市农业科学技术研究院农业环境安全检测研究所（武汉市农业科学技术研究院中心实验室） | Rapid detection method for ochratoxin A |
| CN109444101A (en) * | 2018-12-13 | 2019-03-08 | 四川大学 | Proportional-type aptamer fluorescence probe and its method for detecting ochratoxin A |
| CN109668864A (en) * | 2018-12-14 | 2019-04-23 | 福建中医药大学 | Azotized carbon nano piece couples the ochratoxin A fluorescence detection method of aptamers sensing |
| CN110823846A (en) * | 2018-08-09 | 2020-02-21 | 河北农业大学 | Application of novel fluorescent dye AccuBlue in detection of aptamer |
| CN111896508A (en) * | 2020-07-08 | 2020-11-06 | 西南大学 | Novel method for rapidly detecting ochratoxin A based on nucleic acid dye GeneFinder |
| CN113866405A (en) * | 2021-10-15 | 2021-12-31 | 河南工业大学 | Preparation method of fluorescent aptamer sensor for simultaneously detecting ochratoxin A and aflatoxin B1 |
| CN114121154A (en) * | 2021-10-18 | 2022-03-01 | 江南大学 | Method for improving aptamer specificity and affinity by utilizing molecular design guidance |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101915830A (en) * | 2010-07-19 | 2010-12-15 | 江南大学 | Ochratoxin A fluorescence detection test strip and application thereof |
| US20110306508A1 (en) * | 2008-01-10 | 2011-12-15 | Neoventures Biotechnology Inc. | Method of Mycotoxin Detection |
| CN102517288A (en) * | 2011-11-25 | 2012-06-27 | 国家纳米技术与工程研究院 | Ochratoxin A aptamer and applications thereof |
| CN103055830A (en) * | 2012-12-26 | 2013-04-24 | 华南师范大学 | Preparation method for solid-phase micro-extraction head based on single-stranded DNA aptamer modified graphene oxide coating |
-
2013
- 2013-09-18 CN CN201310429041.8A patent/CN103808701B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110306508A1 (en) * | 2008-01-10 | 2011-12-15 | Neoventures Biotechnology Inc. | Method of Mycotoxin Detection |
| CN101915830A (en) * | 2010-07-19 | 2010-12-15 | 江南大学 | Ochratoxin A fluorescence detection test strip and application thereof |
| CN102517288A (en) * | 2011-11-25 | 2012-06-27 | 国家纳米技术与工程研究院 | Ochratoxin A aptamer and applications thereof |
| CN103055830A (en) * | 2012-12-26 | 2013-04-24 | 华南师范大学 | Preparation method for solid-phase micro-extraction head based on single-stranded DNA aptamer modified graphene oxide coating |
Non-Patent Citations (2)
| Title |
|---|
| 段诺等: "核酸适配体识别-荧光法检测赭曲霉素A", 《分析化学研究报告》 * |
| 陈伶利等: "基于核酸适配体－荧光染料EvaGreenTM 快速检测ATP的研究", 《湖南中医药大学学报》 * |
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| CN104178568B (en) * | 2014-07-25 | 2016-03-30 | 清华大学 | A kind of method based on the target substance in nucleic acid aptamer probe fluorescence sense analyzing and testing sample to be tested |
| CN104178568A (en) * | 2014-07-25 | 2014-12-03 | 清华大学 | Method for detecting target substance in to-be-detected sample based on fluorescent sensing analysis of aptamer probe |
| CN104818279A (en) * | 2015-04-23 | 2015-08-05 | 河南省农业科学院 | Ochratoxin A nucleic acid adapter and application thereof to sample magnetic separation preprocessing |
| CN106546724A (en) * | 2015-09-23 | 2017-03-29 | 中国医学科学院药用植物研究所 | A kind of new method of molecular beacon probe quick detection ochratoxin A |
| CN106124781A (en) * | 2016-07-07 | 2016-11-16 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of method based on aptamers sandwich fluorescent quenching technology for detection demethyl testosterone |
| CN106706737B (en) * | 2016-12-10 | 2019-03-26 | 武汉市农业科学技术研究院农业环境安全检测研究所（武汉市农业科学技术研究院中心实验室） | A kind of ochratoxin A rapid detection method |
| CN106706737A (en) * | 2016-12-10 | 2017-05-24 | 武汉市农业科学技术研究院农业环境安全检测研究所（武汉市农业科学技术研究院中心实验室） | Rapid detection method for ochratoxin A |
| CN110823846A (en) * | 2018-08-09 | 2020-02-21 | 河北农业大学 | Application of novel fluorescent dye AccuBlue in detection of aptamer |
| CN109444101A (en) * | 2018-12-13 | 2019-03-08 | 四川大学 | Proportional-type aptamer fluorescence probe and its method for detecting ochratoxin A |
| CN109444101B (en) * | 2018-12-13 | 2021-03-02 | 四川大学 | Proportional aptamer fluorescent probe and method for detecting ochratoxin A by using same |
| CN109668864A (en) * | 2018-12-14 | 2019-04-23 | 福建中医药大学 | Azotized carbon nano piece couples the ochratoxin A fluorescence detection method of aptamers sensing |
| CN111896508A (en) * | 2020-07-08 | 2020-11-06 | 西南大学 | Novel method for rapidly detecting ochratoxin A based on nucleic acid dye GeneFinder |
| CN113866405A (en) * | 2021-10-15 | 2021-12-31 | 河南工业大学 | Preparation method of fluorescent aptamer sensor for simultaneously detecting ochratoxin A and aflatoxin B1 |
| CN114121154A (en) * | 2021-10-18 | 2022-03-01 | 江南大学 | Method for improving aptamer specificity and affinity by utilizing molecular design guidance |
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