CN107991293A - One kind is used for aflatoxin B1Visible detection method - Google Patents

One kind is used for aflatoxin B1Visible detection method Download PDF

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CN107991293A
CN107991293A CN201711201808.6A CN201711201808A CN107991293A CN 107991293 A CN107991293 A CN 107991293A CN 201711201808 A CN201711201808 A CN 201711201808A CN 107991293 A CN107991293 A CN 107991293A
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probe
aptamers
nano
aflatoxin
nanogold
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谭贵良
刘子雄
王乾蕾
李向丽
郑鸿涛
陈坚
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Zhongshan Institute Of Food And Drug Control
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    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/5308—Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G01N33/54346—Nanoparticles

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Abstract

The invention discloses one kind to be used for aflatoxin B1Visible detection method, by sulfhydrylation aflatoxin B1Aptamers DNA1With the complementary chain dna of sulfhydrylation aptamers2It is connected to form aptamer nanogold probe and complementary strand Nano-Au probe with 13 ± 2nm nanogold respectively, utilizes competition law：When containing target in determinand, target is combined with aptamers, and aptamer nanogold probe and complementary strand Nano-Au probe are in free state, and under high salt concn, nanogold is easily assembled, and color becomes blueness by red；When being free of target in determinand, under high salt concn, nanogold is not assembled, and color is still red；As target concentration increases, nanogold color fades to blueness by red, by measuring uv-vis spectra of the solution in 200~800nm, you can realizes to aflatoxin B in determinand1Quantitative detection.

Description

One kind is used for aflatoxin B1Visible detection method
Technical field
The present invention relates to nano material and biology field, the detection for food security.
Background technology
Aflatoxin is the metabolite of the generations such as aspergillus flavus, aspergillus parasiticus, is broadly divided into aflatoxin B1、B2、 G1、G2And other two kinds of metabolite M1、M2, wherein aflatoxin B1Toxicity it is most strong, there is acute and chronic to humans and animals Toxicity, mutagenicity, carcinogenicity and teratogenesis, its metabolism occurs mainly in liver, therefore the infringement to liver is very big.Aspergillus flavus Toxin is soluble in a variety of organic solvents, such as chloroform, methanol, ethanol, propyl alcohol, a diformamide, water is insoluble in, insoluble in oil Ether, ether and hexane.Aflatoxin B1The food scope of pollution is very wide, mainly peanut, corn, paddy, wheat, peanut oil Deng cereal crops, southern region of China is contaminated the most serious, mainly due to the environment in high temperature, high humidity.Aspergillus flavus poison Plain heat-resisting, 280 DEG C of ability cleavables, therefore generally cook and be difficult to destroy under processing temperature, to ensure the food security of people, China's food Provided in product safety standard：Aflatoxin is not greater than in corn and its product, peanut and its product, peanut oil, corn oil 20μg/kg；Rice, other edible oils are not greater than 10 μ g/kg；Other grains, beans and its product, other shortening nuts, adjust Taste product are not greater than 5 μ g/kg；Special diet food (such as dispensed food for baby, supplementary food for infants) is not greater than 0.5 μg/kg。
It is presently used for detection aflatoxin B1Method mainly have thin-layered chromatography (TLC), high performance liquid chromatography (HPLC) and enzyme-linked immunization (ELISA), these methods respectively have its advantage and disadvantage, and the equipment that TLC methods need is simple, but detects spirit Sensitivity is low；The advantages that HPLC methods high sensitivity, high specificity, but need to carry out complicated pre- place before liquid chromatogram separates Reason, detection and the instrument and equipment for being not suitable for batch samples are expensive；ELISA method has high specificity, the analysis time short etc. Advantage, but stability is poor, and accuracy is not high.Therefore, invention it is a kind of it is easy to operate, cost is low, high specificity, high sensitivity, The short aflatoxin B of detection time1Detection method has great importance.
The content of the invention
The object of the present invention is to provide one kind to be used for aflatoxin B1Visible detection method, modified by aptamers Nanogold forms probe and carries out capture aflatoxin B1, and the optical characteristics for combining nanogold carries out colorimetric, therefore can realize fast Fast ground Visual retrieval aflatoxin B1, therefore the method for the present invention have it is easy to operate, cost is low, high specificity, high sensitivity, The characteristics of detection time is short.
To solve above technical problem, present invention one kind is used for aflatoxin B1Visible detection method, including：
A, by sulfhydrylation aflatoxin B1Aptamers DNA1With the complementary chain dna of sulfhydrylation aptamers2Respectively with 13 ± 2nm nanogold connects to form aptamers-Nano-Au probe and complementary strand-Nano-Au probe；
B, aflatoxin B is carried out by competition law1Visual retrieval, by aptamers-Nano-Au probe, complementary strand- Nano-Au probe and determinand are mixed：
When containing target in determinand, target is combined with aptamers, aptamers-Nano-Au probe and complementary strand-is received Rice Au probe is in free state, and under high salt concn, nanogold is easily assembled, and color becomes au bleu by red；
When being free of target in determinand, aptamers are complementary to the combination of chain base pair complementarity, make aptamers-nanogold Probe and complementary strand-Nano-Au probe are in stable network structure, and under high salt concn, nanogold is not assembled, and color is still For red；
C, with the increase of target concentration in step B, nanogold color gradually becomes blueness by red, by measuring solution In the ultraviolet-visible spectrum of 200~800nm, you can realize to aflatoxin B in determinand1Quantitative detection.
Aflatoxin B1The complementary chain dna of aptamers2Sequence be：
5'SH-TTCACGGTAGCACGCATAGGTGGGGGCAGCTAAAGTCTCC-3'。
When aptamers-Nano-Au probe is prepared in step A, NaCl ageing concentration is 200mmol/L, prepares complementary strand-receive During rice gold, NaCl ageing concentration is 100mmol/L.
Sulfhydrylation aflatoxin B in step A1Aptamers are coupled to form identification capture probe with nanogold, and sulfhydrylation is yellow Aspertoxin B1The complementary strand of aptamers is coupled to form signal probe with nanogold, and aptamers-Nano-Au probe and complementary strand-are received The volume ratio of rice Au probe is 1:1.
It is molten that NaCl is added in step B, after aptamers-Nano-Au probe, complementary strand-Nano-Au probe and determinand mixing Liquid, the NaCl solution concentration are 400mmol/L.
Specifically, the preparation method step of the nanogold is as follows：
(1) container used chloroazotic acid is soaked into about 8h；
The container soaked through chloroazotic acid is cleaned with ultra-pure water, and is dried, the resistivity of the ultra-pure water is more than or equal to 18.2M Ω·cm；
(2) it is 100 that volume ratio is added in container after the drying:1 ultra-pure water and 1% chlorauric acid solution, in oil bath pan In be heated to seething with excitement while stirring, 1% sodium citrate (containing 0.05% citric acid), sodium citrate and gold chloride are added after 10min Volume ratio be 4:1, continue 10~15min of heating to solution and become limpid transparent claret；
(3) stop heating, continue 15~20min of stirring, be cooled to room temperature, that is, obtain the nanometer that particle diameter is 13 ± 2nm Gold grain.
Specifically, the aptamers-Nano-Au probe preparation method step is as follows：
(1) by sulfhydrylation aptamers DNA1With TCEP admixture activations, the molal weight ratio of aptamers and TCEP are 1:100；
(2) by the nanogold at 4 DEG C, the condition of 12000rpm/min first centrifuges 30min, suctions out half volume supernatant Liquid, then vibrate resuspension；
(3) by the aptamers DNA after activation1Add in above-mentioned nano-Au solution, aptamers DNA1With mole of nanogold Mass ratio is 100:1, shaking table is incubated 12~16h under the conditions of 37 DEG C of 150rpm/min, forms DNA1- Nano-Au probe；
(4) lauryl sodium sulfate (SDS) is added into above-mentioned DNA1In-Nano-Au probe, the final concentration of SDS is about 0.01%；
(5) add the NaCl solution of 2mol/L several times in 12h, the final concentration of NaCl is reached 200mmol/L, every time 10~15s of first ultrasound, vibrates mixing before addition NaCl solution after addition；
(6) will be through the DNA after step (5) processing1- Nano-Au probe is aged under the conditions of 37 DEG C of 150rpm/min shaking tables 12h；
(7) by the DNA after ageing1- Nano-Au probe centrifuges 20min under the conditions of 4 DEG C of 8000rpm/min, suctions out supernatant Liquid, the 10mM PBS buffer for adding equivalent are resuspended, and such repeated centrifugation is washed 2 times, to remove what is do not combined with nanogold DNA1, that is, obtain the aptamers-Nano-Au probe.
Specifically, the Na that the formula of the 10mM PBS buffer is 10mmol/L2HPO4, 2mmol/L KH2PO4, PH value is 7.4.
Specifically, the preparation method step of the complementary strand-Nano-Au probe is as follows：
(1) by sulfhydrylation complementary chain dna2With TCEP admixture activations, the molal weight ratio of complementary strand and TCEP are 1:100；
(2) by the nanogold at 4 DEG C, 30min is first centrifuged in the condition of 12000rpm/min, is suctioned out in half volume Clear liquid, then vibrate resuspension；
(3) by the DNA after activation2Add in above-mentioned nano-Au solution, complementary chain dna2With the molal weight ratio of nanogold For 100:1, shaking table is incubated 12~16h under the conditions of 37 DEG C of 150rpm/min, forms DNA2- Nano-Au probe；
(4) lauryl sodium sulfate (SDS) is added into above-mentioned DNA2In-Nano-Au probe, the final concentration of SDS is about 0.01%；
(5) add the NaCl solution of 2mol/L several times in 12h, the final concentration of NaCl solution is reached 100mmol/L, 10~15s of first ultrasound before addition NaCl solution every time, vibrates mixing after addition；
(6) will be through the DNA after step (5) processing2- Nano-Au probe is aged under the conditions of 37 DEG C, 150rpm/min shaking tables 12h；
(7) by the DNA after ageing2- Nano-Au probe centrifuges 20min under the conditions of 4 DEG C of 8000rpm/min, suctions out supernatant Liquid, the 10mM PBS buffer for adding equivalent are resuspended, and such repeated centrifugation is washed 2 times, to remove what is do not combined with nanogold DNA2, that is, obtain the complementary strand-Nano-Au probe.
Specifically, the Na that the formula of the 10mM PBS buffer is 10mmol/L2HPO4, 2mmol/L KH2PO4, PH value is 7.4.
Specifically, 95 DEG C of aptamers-Nano-Au probe, complementary strand-Nano-Au probe water-bath 5min, then will be suitable Ligand-Nano-Au probe, complementary strand-Nano-Au probe, determinand by volume 1:1:1 is incubated 1h under the conditions of 37 DEG C.
Specifically, NaCl solution is added after aptamers-Nano-Au probe, complementary strand-Nano-Au probe, determinand mixing, The concentration of the NaCl solution is 400mmol/L.
Testing principle：DNA1With DNA2Be complete complementary pairing base sequence, DNA1It can identify and capture target substance Huang Aspertoxin B1, complementary chain dna2And aflatoxin B1Competition and aptamers DNA1With reference to.It is malicious when containing aspergillus flavus in determinand Plain B1When, DNA1With aflatoxin B1With reference to free state, under certain salinity, nanometer gold surface is presented in nanogold Negative electrical charge shielded, repulsive force between nano particle weakens, and clustering phenomena occurs, since the color of nanogold has electrical distance Dependency characteristic, therefore the color of nanogold changes, aflatoxin B in determinand1Concentration it is different, nanogold aggregation Degree is also different, so as to cause the color of nano-Au solution also different.When there is no aflatoxin B in determinand1When, DNA1With DNA2Complementary hybridization, forms stable network structure, and under certain salinity, the distance between nano Au particle is not drawn Closely, therefore the color of nano-Au solution does not change, and aflatoxin B is realized according to the change of nano-Au solution color1 Visual retrieval.
Compared with prior art, amplifying nucleic acid aptamers of the present invention are section of DNA sequence, are to utilize in-vitro screening skill The Fas lignand system evolution technology (SELEX) of art --- index concentration, the oligonucleotide fragment obtained from nucleic acid molecule libraries, is A kind of new identification molecule, has that affinity height, high specificity, molecular weight are small, chemically synthesize, stability is good, non-toxic etc. Advantage.And nanogold has very unique optical characteristics, when the distance between gold nano grain furthers, nanogold face can be caused The change of color.The present invention realizes aflatoxin B using the optical characteristics of nanogold and high specific, the sensitivity of aptamers1 Quick visualization detection.And the present invention is to aflatoxin B1Detection be limited to 0.1ng/ml, in 0.1~1000ng/ml models Enclose interior linear good (R2=0.99158), moreover, aptamers-probe and complementary strand-Nano-Au probe it is easily prepared and preserve, Detection is completed in 1h, and easy to operate, can visually distinguish the color change of nanogold, it is not necessary to complex instrument into Row detection, easy to live detection in real time.
Brief description of the drawings
Fig. 1 is detection principle diagram of the present invention.
Fig. 2 is the transmission electron microscope picture of nanogold in the present invention.
Fig. 3 is nanogold of the present invention and aptamers DNA1, complementary chain dna2The ultraviolet-ray visible absorbing collection of illustrative plates of modified nano gold.
Fig. 4 is aflatoxin B in the present invention1Examination criteria curve map.
Embodiment
Below in conjunction with the accompanying drawings and specific embodiment is described in detail the present invention.
Embodiment 1：
1. the mercapto-modified aptamers DNA of synthesis1, and with the complementary chain dna of aptamers complementary pairing2(purchased from Shanghai life work Bioengineering limited company).
(1) aptamers DNA1Sequence is：
5'SH-AGCAGCACAGAGGTCAGATGGTGCTATCATGCGCTCAATGGGAGACTTTAGCTGCCCCCACCTA TGCGTGCTACCGTGAA-3'；
(2) complementary chain dna2Sequence is：
5'SH-TTCACGGTAGCACGCATAGGTGGGGGCAGCTAAAGTCTCC-3'；
2. prepare nanogold.
(1) used container chloroazotic acid is soaked into 12h, the chloroazotic acid is 3 by volume ratio:1 concentrated hydrochloric acid and concentrated nitric acid Prepare；
(2) container soaked through chloroazotic acid is cleaned up with ultra-pure water, and is dried；
(3) ultra-pure water and 0.5mL, 1% chlorauric acid solution of 50mL and resistivity >=18.2M Ω are added in a reservoir, are used Rotor stirs evenly, and boils 10min, 2mL, 1% sodium citrate is added, wherein containing 0.05% citric acid, solution then gradually becomes Aubergine, continues to heat 10min；
(4) stop heating, continue to stir 20min, cool down at room temperature, obtain the nano-Au solution of claret, pass through purple Outside-visible absorbance collection of illustrative plates and transmission electron microscope characterize prepared nano-Au solution, and characterization collection of illustrative plates is shown in Fig. 2.
3. prepare aptamers-Nano-Au probe.
(1) adapted dna of 20 μ L10 μm ol/L is taken1In 1.5mL centrifuge tubes, the TCEP and 20 of 20 μ L 1mmol/L is added The PBS of μ L, pH=7.4,10mmol/L, admixture activation 1h；
(2) 2mL nanogold is taken to be placed in 2mL centrifuge tubes, 4 DEG C, 12000rpm/min centrifugation 30min, suction out 1mL supernatants Liquid, then resuspension is vibrated, obtain the nano-Au solution of 2 times of concentration；
(3) aptamers activated are added in the nano-Au solution of 2 times of concentration, 37 DEG C of shaking tables are incubated 12h；
(4) SDS of 10.6 μ L1% is added in aptamers-Nano-Au probe, makes SDS concentration up to 0.01%；
(5) point 6 107 μ L of addition, the NaCl solutions of 2mol/L in 12h, add 17.8 μ L, make NaCl solution most every time Final concentration reaches 200mmol/L, adds first ultrasound 10s before NaCl solution every time, mixing is vibrated after addition, in 37 DEG C, 150rpm/ 12h is aged under the conditions of min shaking tables；
(6) aptamers-Nano-Au probe after being aged centrifuges 20min in 4 DEG C, 8000rpm/min, suctions out supernatant, then The PBS buffer for adding 10mmol/L, pH=7.4 of equivalent is resuspended, and such repeated centrifugation is washed 2 times, with remove not with nanometer The aptamers that gold combines, obtained aptamers-Nano-Au probe are spare under the conditions of being stored in 4 DEG C.
4. complementary strand-Nano-Au probe.
(1) adapted dna of 20 10 μm of ol/L of μ L is taken2In 1.5mL centrifuge tubes, add 20 μ L 1mmol/L TCEP and The PBS of 20 μ L, pH=7.4,10mmol/L, admixture activation 1h；
(2) take 2mL nanogold to be placed in 2mL centrifuge tubes, in 4 DEG C, 12000rpm/min centrifugation 30min, suction out 1mL supernatants Liquid, then resuspension is vibrated, obtain the nano-Au solution of 2 times of concentration；
(3) aptamers activated are added in the nano-Au solution of 2 times of concentration, 37 DEG C of shaking tables are incubated 12h；
(4) SDS of 10.6 μ L 1% is added in aptamers-Nano-Au probe, makes SDS concentration up to 0.01%；
(5) point 6 53.5 μ L of addition, the NaCl solutions of 2mol/L in 12h, add 8.9 μ L, make NaCl solution most every time Final concentration reaches 100mmol/L, adds first ultrasound 10s before NaCl solution every time, mixing is vibrated after addition, in 37 DEG C, 150rpm/ 12h is aged under the conditions of min shaking tables；
(6) aptamers-Nano-Au probe after being aged centrifuges 20min in 4 DEG C, 8000rpm/min, suctions out supernatant, then The PBS buffer for adding 10mmol/L, pH=7.4 of equivalent is resuspended, and such repeated centrifugation is washed 2 times, with remove not with nanometer The aptamers that gold combines, obtained aptamers-Nano-Au probe are spare under the conditions of being stored in 4 DEG C.
5. aflatoxin B1Detection.
Embodiment 1：
Aptamers-the Nano-Au probe prepared and complementary strand-Nano-Au probe are placed in 95 DEG C of heat treatment in water-bath 5min, then takes the Huang of 100 μ L aptamers-Nano-Au probe, 100 μ L complementary strands-Nano-Au probe, 100 μ L various concentrations bent Mould toxin B1Solution (0ng/ml, 0.1ng/ml, 1.0ng/ml, 10ng/ml, 100ng/ml, 1000ng/ml), be placed in 1.5mL from Mixed in heart pipe, 1h is incubated in 37 DEG C of shaking tables, then added the NaCl solution of 75 μ L2mol/L, make NaCl solution ultimate density Reach 400mmol/L, react 2min；Solution colour change is observed, and UV-vis scannings are carried out in 200~800nm.According to extinction Angle value (ratio of A650/A525) and aflatoxin B1Concentration draw aflatoxin B1Standard curve, referring to Fig. 4.
Embodiment 2：
Aflatoxin B in peanut butter actual sample1Detection and recovery of standard addition detection.
(1) sample pretreatment is detected：Weigh 4.0g peanut butter to be placed in 50mL centrifuge tubes, the methanol for adding 10mL70% carries Solution is taken, in acutely shaking 10min on oscillator, is filtered after standing 30min with quantitative filter paper, takes 0.1mL filtrates, add 1.9mL ultra-pure waters are diluted, and obtain solution to be checked with spare.
(2) aflatoxin B in the method for the present invention determination sample is utilized1Content, experimental procedure is with embodiment 1, as a result It is shown in Table one.
(3) aflatoxin B obtained with step (2)1Concentration data is background values, adds various concentrations thereto AFB1Standard items, aflatoxin B is detected also with the method for the present invention again1Content, obtains detected value.Rate of recovery %=(inspections Measured value-background values)/additive amount × 100%.The rate of recovery is can see 95.2%~109.6% from one data of table, illustrates this hair Bright stabilization, it is sensitive, accurately, suitable for the detection of beer actual sample.
Table one：Aflatoxin B in peanut butter actual sample1Detection and recovery of standard addition
Note：ND is not detect.

Claims (5)

1. one kind is used for aflatoxin B1Visible detection method, it is characterised in that：
A, by sulfhydrylation aflatoxin B1Aptamers DNA1With the complementary chain dna of sulfhydrylation aptamers2Received respectively with 13 ± 2nm Rice gold connection forms aptamers-Nano-Au probe and complementary strand-Nano-Au probe；
B, aflatoxin B is carried out by competition law1Visual retrieval, by aptamers-Nano-Au probe, complementary strand-nanogold Probe and determinand are mixed：
When containing target in determinand, target is combined with aptamers, makes aptamers-Nano-Au probe and complementary strand-nanogold Probe is in free state, and under high salt concn, nanogold is easily assembled, and color becomes au bleu by red；
When being free of target in determinand, aptamers are complementary to the combination of chain base pair complementarity, make aptamers-Nano-Au probe With complementary strand-Nano-Au probe in stable network structure, under high salt concn, nanogold is not assembled, and color is still to be red Color；
C, as the increase of target concentration in step B, nanogold color gradually become blueness by red, existed by measuring solution The ultraviolet-visible spectrum of 200~800nm, you can realize to aflatoxin B in determinand1Quantitative detection.
2. a kind of according to claim 1 be used for aflatoxin B1Visible detection method, it is characterised in that：Aspergillus flavus Toxin B1The complementary chain dna of aptamers2Sequence be：
5'SH-TTCACGGTAGCACGCATAGGTGGGGGCAGCTAAAGTCTCC-3'。
3. a kind of according to claim 1 be used for aflatoxin B1Visible detection method, it is characterised in that：In step A When preparing aptamers-Nano-Au probe, NaCl solution ageing concentration is 200mmol/L, when preparing complementary strand-nanogold, NaCl Solution ageing concentration is 100mmol/L.
4. a kind of according to claim 1 be used for aflatoxin B1Visible detection method, it is characterised in that：In step A Sulfhydrylation aflatoxin B1Aptamers are coupled to form identification capture probe with nanogold, sulfhydrylation aflatoxin B1Aptamers Complementary strand and nanogold be coupled to form signal probe, the volume ratio of aptamers-Nano-Au probe and complementary strand-Nano-Au probe For 1:1.
5. a kind of according to claim 1 be used for aflatoxin B1Visible detection method, it is characterised in that：Step B In, NaCl solution is added after aptamers-Nano-Au probe, complementary strand-Nano-Au probe and determinand mixing, the NaCl solution is dense Spend for 400mmol/L.
CN201711201808.6A 2017-11-27 2017-11-27 One kind is used for aflatoxin B1Visible detection method Pending CN107991293A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109030442A (en) * 2018-08-01 2018-12-18 岭南师范学院 A kind of detection method of detection probe and aflatoxin for detecting aflatoxin AFB1
CN113720794A (en) * 2021-06-11 2021-11-30 海南大学 Method for sensing and detecting mycotoxin in rice by using gold nanoparticle-based colorimetric aptamer

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