CN101059517A - Method for checking aflatoxin B1 in agricultural by-product - Google Patents
Method for checking aflatoxin B1 in agricultural by-product Download PDFInfo
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- CN101059517A CN101059517A CN 200710035069 CN200710035069A CN101059517A CN 101059517 A CN101059517 A CN 101059517A CN 200710035069 CN200710035069 CN 200710035069 CN 200710035069 A CN200710035069 A CN 200710035069A CN 101059517 A CN101059517 A CN 101059517A
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Abstract
The invention discloses a method for checking aflatoxin B1 in farm and side line products, comprising that (1), preparing aflatoxin B1 piezoelectric immunity sensor on the surface of a quartz crystal micro scale gold electrode, (2), preparing antipest IgG antibody of nanometer gold mark, (3), preparing standard working curvature, (4), checking sample. The invention uses the sensitivity of piezoelectric crystal on quality change, combined with a biological recognize system (antigen and antibody character combination) to form an automatic analyzing checking system, to check the aflatoxin B1 density in farm and side line products. The invention uses the two-anti and one-anti immunity recognition of nanometer gold mark to realize the surface combination of macromolecule antibody, to amplify the quality response, to realize the high-sensitivity quantitative test of aflatoxin B1.
Description
Technical field
The invention belongs to field of biological detection, be specifically related to a kind of method that detects aflatoxin B1 in the agricultural byproducts.
Background technology
Aflatoxin B1 is mould etc. the metabolic product of aspergillus flavus, aspergillus parasiticus, Tequ, is I class carcinogenic substance, is the extremely strong mycotoxin of a kind of toxicity.Because its harm to human body, strict regulation has all been made to the maximum permissible concentration of aflatoxin B1 in the agricultural byproducts by each state, and the highly sensitive analysis means of setting up aflatoxin B1 is significant to agricultural byproducts supervision and detection.The aflatoxin B1 detection method of using has now: thin-layered chromatography, high performance liquid chromatography, immune affinity column fluorescence spectrophotometry, enzyme linked immunosorbent assay etc.These detection methods are complex steps mostly, needs specific apparatus, and length consuming time is not suitable for on-site quick screening.Therefore, need a kind of analysis speed fast, highly sensitive, but the detection method of on-site quick screening.
Summary of the invention
The technical problem to be solved in the present invention is to overcome the deficiencies in the prior art, and a kind of method that detects aflatoxin B1 in the agricultural byproducts is provided.The present invention is used for the detection to the agricultural byproducts aflatoxin B1, and it is low to have an analysis cost, and analysis speed is fast, and is highly sensitive, reaches AFB in China's agricultural byproducts
1Limit standard.Analytical instrument is easy to carry, and can realize on-the-spot supervision and detect.
In order to achieve the above object, the method for aflatoxin B1 may further comprise the steps in the detection agricultural byproducts provided by the invention:
1, prepares AFB in the QCM (Quartz Crystal Microbalance) gold electrode surfaces
1Piezoelectric immunosensor;
2, the sheep anti-mouse igg antibody of preparation nano gold mark (two is anti-);
3, make standard working curve:
Get the AFB of variable concentrations
1Standard solution adds the AFB of conventional concentration amounts
1Monoclonal antibody adds bovine serum albumin(BSA) again, and mixing is got convention amount and dripped to the AFB that has prepared
1Piezoelectric immunosensor reacts 30min in 37 ℃ of environment; Then add nano gold mark antibody (two is anti-), detect the frequency change of the little balance of piezoelectric quartz crystal in real time; Change the AFB of normal concentration
1, measure a series of AFBs by above-mentioned steps
1Standard solution, with piezoelectric quartz crystal little balance frequency change value and AFB
1Concentration drawing standard working curve;
4, sample detection:
Get the sample solution of handling well, add the AFB of conventional concentration amounts
1Monoclonal antibody adds bovine serum albumin(BSA) again, and mixing is got convention amount and dripped to the AFB that has prepared
1Piezoelectric immunosensor reacts 30min in 37 ℃ of environment; Then add nano gold mark antibody (two is anti-), detect the frequency change of the little balance of piezoelectric quartz crystal in real time; Detect the frequency change of the little balance of piezoelectric quartz crystal in real time, the recording frequency changing value; The standard working curve contrast of frequency change value and the drafting of the 3rd step is obtained AFB in the sample
1Numerical value.
AFB described in the inventive method step 1
1Piezoelectric immunosensor can be prepared by following steps:
1) half Guang ammonia self assembly of QCM (Quartz Crystal Microbalance) gold electrode surfaces: gold electrode is immersed in the half Guang ammonia spirit of 2.0mmol/L, standing over night, take out gold electrode, gold electrode surfaces is cleaned three times with pure water, promptly obtains the fine and close uniform half Guang ammonia self-assembled monolayer of one deck in gold electrode surfaces;
2) QCM (Quartz Crystal Microbalance) surface antibody immobilization: with self assembly the crystal oscillator of half Guang ammonia self-assembled monolayer immerse in 5.0% (V/V) glutaraldehyde solution, left standstill 1～1.5 hour, take out gold electrode, gold electrode surfaces is cleaned three times with pure water, will contain 10.0 μ g/mL AFBs again
1Bovine serum albumin(BSA) cross-linking agent solution 20 μ L drip to electrode surface, left standstill 1～1.5 hour, electrode surface cleans three times with pure water, getting massfraction and be 1% bovine serum albumin solution 50 μ L drips to electrode surface, leave standstill sealing 0.5 hour, electrode surface cleans three times with pure water, promptly obtains AFB
1Piezoelectric immunosensor.
The preparation of the sheep anti-mouse igg antibody of nano gold mark described in the inventive method step 2 can be carried out by the following method:
Get nm of gold colloid 250 μ l, regulating pH with 0.1mol/L sodium carbonate-sodium bicarbonate buffer liquid is 9.0; Get sheep anti-mouse igg antibody 5 μ l, dropwise join in the aurosol, strong agitation 10min～20min then in incubated at room 1～1.5h, tentatively makes nano gold mark antibody; Add massfraction and be 5.0% bovine serum albumin(BSA) (BSA) solution in the nano gold mark antibody-solutions, making BSA final mass mark is 1.0%, sealing nano gold mark antibody; The nanometer labelled antibody is inserted in the centrifuge tube of 1.5mL, with the speed refrigerated centrifuge 10min of 15000r/min, the control centrifuging temperature is 4 ℃; Discard supernatant liquor, (PBS pH=7.4) washs the kermesinus sediment 2 times with containing the phosphate buffer solution that massfraction is 0.1%BSA; It is in 0.1% the BSA-PBS damping fluid that this sediment is suspended in massfraction again, makes nano gold mark antibody.
Detection principle of the present invention:
Detection method of the present invention is based on utilizes the susceptibility of piezoelectric crystal to mass change, and a kind of automated analysis detection system that forms in conjunction with biological recognition system (antigen and antibody specific in conjunction with) detects aflatoxin B1 concentration in the agricultural byproducts.Among the present invention, AFB
1Holoantigen is modified the little balance sensing interface of piezoelectric quartz crystal, forms AFB
1Piezoelectric immunosensor.AFB in antibody and the sample
1And immobilized AFB
1The competition effect of holoantigen, simultaneously, utilize nano gold mark two anti-and anti-immunity identifications (two anti-be on an anti-basis, to prepare, anti-the specific recognition effect arranged to one), realize the surface combination of big molecular antibody, further amplify mass-basis response, the quality amplification can reach millions of times, and (the little balance of piezoelectric crystal itself is exactly the sensing device to mass-sensitive, because the quality of nm of gold will be far longer than the quality of antibody or antigen, get the AFB of variable concentrations
1Standard solution adds the AFB of conventional concentration amounts
1Monoclonal antibody adds bovine serum albumin(BSA) again, and mixing is got convention amount and dripped to the AFB that has prepared
1Piezoelectric immunosensor reacts 30min in 37 ℃ of environment, obviously, and AFB
1And AFB
1Monoclonal antibody has been reacted earlier, if AFB in the sample
1Many, because most of AFB
1Antibody with sample in AFB
1Reacted, be connected to the AFB on crystal oscillator surface so
1Antibody is just few, so nano gold mark two resist up also just few.Therefore, this experiment AFB in its enlargement factor and the sample in fact
1Content be inversely proportional to, that is to say the AFB in the sample
1Few more, the antibody that is connected to the crystal oscillator surface so is many more on the contrary, so nano gold mark two resist up also many more, it is obvious more that quality is amplified.Just have high sensitivity Just because of this.), in 30min, can finish reaction, the recording frequency changing value.
The present invention has mainly solved the serial problem of aflatoxin B1 fast detecting: because AFB
1Molecular weight is little, and piezoelectric quality response is little, and the present invention utilizes and analyzes in the sample antigen and immobilized antigen the competition effect of free antibodies is realized the surface combination of big molecular antibody, and response improves the quality.Simultaneously, utilize two anti-and anti-immunity identifications of nano gold mark, further amplify mass-basis response, realize AFB
1Highly sensitive quantitative measurement.
The present invention adopts AFB
1-bovine serum albumin(BSA) (BSA) cross-linking agent (being holoantigen) carries out haptenic fixing, improves area load amount and fixed efficiency, simplifies the sensor production program.The optimization of condition by experiment, this sensor is to AFB
1Frequency response fairly obvious, but other mycotoxins are not almost had frequency response.The effect of good restraining non-specific adsorption has been played on the immune sensing surface of finishing aflatoxin B1 and bovine serum albumin(BSA) sealing.Sensor can soak 30min～40min regeneration with 6.0mol/L urea.Regeneration back sensor can use repeatedly and not have a remarkable loss of sensitivity.
The AFB of preparation
1It is simple that piezoelectric immunosensor detects step, is no more than 1 hour detection time, and the regeneration of sensor interface is easy, can repeatedly use repeatedly.The detection sensitivity height, lower limit can reach 0.50ng/mL, and the analysis precision relative error is in 8.0%.It is portable and cheap to detect required QCM (Quartz Crystal Microbalance) instrument.
The present invention can utilize common immunological assay reagents, need not special marking, obtains the raising greatly of mass-basis response, has significantly improved the sensitivity of sensor.Analysis cost is low, and analysis speed is fast, and is highly sensitive, can reach AFB in China's agricultural byproducts
1Limit standard.Instrument can be portable, can realize on-the-spot supervision and detect.
Embodiment
Embodiment:
1, preparation AFB
1Piezoelectric immunosensor:
1), half Guang ammonia self assembly of QCM (Quartz Crystal Microbalance) gold electrode surfaces: gold electrode is immersed in the half Guang ammonia spirit of 2.0mmol/L, standing over night, take out gold electrode, gold electrode surfaces is cleaned three times with pure water, promptly obtains the fine and close uniform half Guang ammonia self-assembled monolayer of one deck in gold electrode surfaces;
2), QCM (Quartz Crystal Microbalance) surface antibody immobilization: with self assembly the crystal oscillator of half Guang ammonia self-assembled monolayer immerse in 5.0% (V/V) glutaraldehyde solution, left standstill 1～1.5 hour, take out gold electrode, gold electrode surfaces is cleaned three times with pure water, will contain 10.0 μ g/mL AFBs again
1Bovine serum albumin(BSA) cross-linking agent solution 20 μ L drip to electrode surface, left standstill 1～1.5 hour, electrode surface cleans three times with pure water, getting massfraction and be 1% bovine serum albumin solution 50 μ L drips to electrode surface, leave standstill sealing 0.5 hour, electrode surface cleans three times with pure water, promptly obtains AFB
1Piezoelectric immunosensor.
2, the sheep anti-mouse igg antibody of preparation nano gold mark:
Get nm of gold colloid 250 μ l, regulating pH with 0.1mol/L sodium carbonate-sodium bicarbonate buffer liquid is 9.0; Get sheep anti-mouse igg antibody 5 μ l, dropwise join in the aurosol, strong agitation 10min～20min in incubated at room 1～1.5 hour, tentatively makes nano gold mark antibody then; Add massfraction and be 5.0% bovine serum albumin(BSA) (BSA) solution in the nano gold mark antibody-solutions, making BSA final mass mark is 1.0%, sealing nano gold mark antibody; The nanometer labelled antibody is inserted in the centrifuge tube of 1.5mL, with the speed refrigerated centrifuge 10min of 15000r/min, the control centrifuging temperature is 4 ℃; Discard supernatant liquor, (PBS pH=7.4) washs the kermesinus sediment 2 times with containing the phosphate buffer solution that massfraction is 0.1%BSA; It is in 0.1% the BSA-PBS damping fluid that this sediment is suspended in massfraction again, the experiment that makes required nano gold mark antibody, this nano gold mark antibody can be preserved more than 6 months in 4 ℃ of refrigerators.
3, measure AFB
1The drafting of concentration standard working curve:
AFB with variable concentrations
1Mix respectively and contain the 100.0ng/mL AFB
1Monoclonal antibody, adding massfraction again is 1.0% bovine serum albumin(BSA), mixing is got 20 μ L respectively and is dripped to the AFB for preparing
130min is reacted in 37 ℃ of environment in the piezoelectric immunosensor surface; Behind surperficial three or four times of pure water cleaning sensor, the sheep anti-mouse igg antibody that adds the 10.0nmol/L nano gold mark detects the frequency change of the little balance of piezoelectric quartz crystal in real time, observes the initial period frequency and descends very fast, in 30min, can finish reaction, the recording frequency changing value; Measure a series of AFBs by above-mentioned steps
1Standard solution such as 0.70ng/mL, 5.0ng/mL, 10.0ng/mL, 20.0ng/mL, 50.0ng/mL, 100.0ng/mL, 130.0ng/mL is with piezoelectric quartz crystal little balance frequency change value and AFB
1Concentration drawing standard working curve, this working curve is at the 0.70-130.0ng/mL AFB
1Linear in the concentration range, be limited to 0.50ng/mL under detecting.
4, AFB in the testing sample
1Detection:
Accurately take by weighing testing sample 10.0～250.0 grams, add 0.2mol/L phosphate buffered solution 150～250mL, homogeneous becomes homogenate; Cross leaching supernatant liquor 10～30mL, use methyl alcohol: water (60: 40, V/V) solution 10～30mL leaves standstill and extracts 10～20min, dilutes with 20～50mL water; Get sample 50～200 μ L that handle well, mix and contain 100.0～300.0ng/mL aflatoxin B1 monoclonal antibody, adding massfraction again is 1.0% bovine serum albumin(BSA), mixing, get 10～20 μ L and drip, react 30～50min in 37 ℃ of environment to the aflatoxin B1 piezoelectric immunosensor surface for preparing; Behind surperficial three or four times of pure water cleaning sensor, add the sheep anti-mouse igg antibody of 10.0～20.0nmol/L nano gold mark, detect the frequency change of the little balance of piezoelectric quartz crystal in real time; The observed frequency situation of change can be finished reaction at 30～40min; The recording frequency changing value; The standard working curve contrast of frequency change value and the drafting of the 3rd step is obtained AFB in the sample
1Numerical value.
Detect example 1:
To the AFB in the milk
1The detection of the recovery: get and measured milk (NF AFB
1) 20mL, the AFB of adding 2.0ng/mL
1Use methyl alcohol: (60: 40, V/V) solution 20mL left standstill and extracts 10min water, with the dilution of 20mL water.Get above-mentioned sample 100 μ L, add the 100.0ng/mL AFB
1Monoclonal antibody, massfraction are 1.0% bovine serum albumin(BSA), mixing.Getting wherein, 20 μ L drip to the aflatoxin B1 piezoelectric immunosensor surface for preparing.React 30min in 37 ℃ of environment.Behind surperficial three times of pure water cleaning sensor, add the sheep anti-mouse igg antibody of 10.0nmol/L nano gold mark, detect the frequency change of the little balance of piezoelectric quartz crystal in real time.It is very fast to observe initial period frequency decline, can finish reaction in 30 minutes.The recording frequency changing value.Calculate AFB according to working curve
1Concentration.Measure AFB five times
1Concentration obtain its as a result mean value be 1.91 ± 0.15ng/mL.Coincide with five enzyme linked immunosorbent assay result 1.84 ± 0.13ng/mL.The result shows, the recovery 85.5%～96.5% of this sensor, and reaching can be to AFB in the milk
1Carry out fast detecting.
Detect example 2:
Detection to the aflatoxin B1 in the mouldy peanut sample: get mouldy peanut sample 250.0g, add 0.2mol/L phosphate buffered solution 200mL, homogeneous becomes homogenate.Cross leaching supernatant liquor 20mL, use methyl alcohol: (60: 40, V/V) solution 20mL left standstill and extracts 10min water, with the dilution of 20mL water.Get above-mentioned sample 100 μ L, add 100.0ng/mL aflatoxin B1 monoclonal antibody, massfraction is 1.0% bovine serum albumin(BSA), mixing.Getting wherein, 20 μ L drip to the AFB for preparing
1The piezoelectric immunosensor surface.React 30min in 37 ℃ of environment.Behind surperficial three times of pure water cleaning sensor, add the sheep anti-mouse igg antibody of 10.0nmol/L nano gold mark, detect the frequency change of the little balance of piezoelectric quartz crystal in real time.It is very fast to observe initial period frequency decline, can finish reaction in 30 minutes.The recording frequency changing value.Measure for five times and calculate AFB in the corresponding test solution according to working curve
1Concentration mean value is 63.3 ± 2.0ng/mL.Coincide with five enzyme linked immunosorbent assay result 61.8 ± 3.2ng/mL.The result shows that this sensor can carry out fast detecting to aflatoxin B1 in the peanut.
Detect example 3:
To the AFB in the mouldy rice sample
1Detection: get mouldy rice sample 200.0g, add 0.2mol/L phosphate buffered solution 200mL, homogeneous becomes homogenate.Cross leaching supernatant liquor 20mL, use methyl alcohol: (60: 40, V/V) solution 20mL left standstill and extracts 10min water, with the dilution of 20mL water.Get above-mentioned sample 100 μ L, add 100.0ng/mL aflatoxin B1 monoclonal antibody, massfraction is 1.0% bovine serum albumin(BSA), mixing.Getting wherein, 20 μ L drip to the AFB for preparing
1The piezoelectric immunosensor surface.React 30min in 37 ℃ of environment.Behind surperficial three times of pure water cleaning sensor, add the sheep anti-mouse igg antibody of 10.0nmol/L nano gold mark, detect the frequency change of the little balance of piezoelectric quartz crystal in real time.It is very fast to observe initial period frequency decline, can finish reaction in 30 minutes.The recording frequency changing value.Measure for five times and calculate AFB in the corresponding test solution according to working curve
1Concentration mean value is 30.1 ± 1.5ng/mL.Coincide with five enzyme linked immunosorbent assay result 28.4 ± 2.5ng/mL.The result shows that this sensor can be to AFB in the rice
1Carry out fast detecting.
Claims (3)
1, a kind of method that detects aflatoxin B1 in the agricultural byproducts is characterized in that may further comprise the steps:
1) prepares AFB in the QCM (Quartz Crystal Microbalance) gold electrode surfaces
1Piezoelectric immunosensor;
2) sheep anti-mouse igg antibody of preparation nano gold mark;
3) make standard working curve:
Get the AFB of variable concentrations
1Standard solution adds the AFB of conventional concentration amounts
1Monoclonal antibody adds bovine serum albumin(BSA) again, and mixing is got convention amount and dripped to the AFB that has prepared
1Piezoelectric immunosensor reacts 30min in 37 ℃ of environment; Then add nano gold mark antibody (two is anti-), detect the frequency change of the little balance of piezoelectric quartz crystal in real time; Change the AFB of normal concentration
1, measure a series of AFBs by above-mentioned steps
1Standard solution, with piezoelectric quartz crystal little balance frequency change value and AFB
1Concentration drawing standard working curve;
4) sample detection:
Get the sample solution of handling well, add the AFB of conventional concentration amounts
1Monoclonal antibody adds bovine serum albumin(BSA) again, and mixing is got convention amount and dripped to the AFB that has prepared
1Piezoelectric immunosensor reacts 30min in 37 ℃ of environment; Then add nano gold mark antibody (two is anti-), detect the frequency change of the little balance of piezoelectric quartz crystal in real time; Detect the frequency change of the little balance of piezoelectric quartz crystal in real time, the recording frequency changing value; With frequency change value and the 3rd) the step standard working curve contrast of drawing obtains AFB in the sample
1Numerical value.
2, the method for aflatoxin B1 in the detection agricultural byproducts according to claim 1 is characterized in that AFB described in the step 1)
1Piezoelectric immunosensor is prepared by following steps:
(1) half Guang ammonia self assembly of QCM (Quartz Crystal Microbalance) gold electrode surfaces: gold electrode is immersed in the half Guang ammonia spirit of 2.0mmol/L, standing over night, take out gold electrode, gold electrode surfaces is cleaned three times with pure water, promptly obtains the fine and close uniform half Guang ammonia self-assembled monolayer of one deck in gold electrode surfaces;
(2) QCM (Quartz Crystal Microbalance) surface antibody immobilization: with self assembly the crystal oscillator of half Guang ammonia self-assembled monolayer immerse in 5.0% (V/V) glutaraldehyde solution, left standstill 1～1.5 hour, take out gold electrode, gold electrode surfaces is cleaned three times with pure water, will contain 10.0 μ g/mL AFBs again
1Bovine serum albumin(BSA) cross-linking agent solution 20 μ L drip to electrode surface, left standstill 1～1.5 hour, electrode surface cleans three times with pure water, getting massfraction and be 1% bovine serum albumin solution 50 μ L drips to electrode surface, leave standstill sealing 1～1.5 hour, electrode surface cleans three times with pure water, promptly obtains AFB
1Piezoelectric immunosensor.
3, the method for aflatoxin B1 in the detection agricultural byproducts according to claim 1 is characterized in that step 2) described in the preparation of sheep anti-mouse igg antibody of nano gold mark carry out by the following method:
Get nm of gold colloid 250 μ l, regulating pH with 0.1mol/L sodium carbonate-sodium bicarbonate buffer liquid is 9.0; Get sheep anti-mouse igg antibody 5 μ l, dropwise join in the aurosol, strong agitation 10min～20min then in incubated at room 1～1.5h, tentatively makes nano gold mark antibody; Add massfraction and be 5.0% bovine serum albumin(BSA) (BSA) solution in the nano gold mark antibody-solutions, making BSA final mass mark is 1.0%, sealing nano gold mark antibody; The nanometer labelled antibody is inserted in the centrifuge tube of 1.5mL, with the speed refrigerated centrifuge 10min of 15000r/min, the control centrifuging temperature is 4 ℃; Discard supernatant liquor, (PBS pH=7.4) washs the kermesinus sediment 2 times with containing the phosphate buffer solution that massfraction is 0.1%BSA; It is in 0.1% the BSA-PBS damping fluid that this sediment is suspended in massfraction again, makes nano gold mark antibody.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2007100350698A CN101059517B (en) | 2007-06-06 | 2007-06-06 | Method for checking aflatoxin B1 in agricultural by-product |
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| CN2007100350698A CN101059517B (en) | 2007-06-06 | 2007-06-06 | Method for checking aflatoxin B1 in agricultural by-product |
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| CN101059517B CN101059517B (en) | 2011-05-11 |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101216450B (en) * | 2008-01-16 | 2010-09-01 | 暨南大学 | Biosensor electrode for detecting aspergillus flavus toxin B1 and method for making same |
| CN102213687A (en) * | 2010-04-09 | 2011-10-12 | 中国科学院海洋研究所 | Impedance sensor for detecting microbes quickly and construction method thereof |
| CN102692513A (en) * | 2012-06-26 | 2012-09-26 | 长沙理工大学 | Piezoelectric immunosensing chip and device for detecting oncogene C-myc recombinant protein |
| CN103196973A (en) * | 2013-03-13 | 2013-07-10 | 济南大学 | Method for preparing seven-channel aflatoxin immunosensor and application |
| CN103196984A (en) * | 2013-03-13 | 2013-07-10 | 济南大学 | Preparation method and application of sensor for simultaneous detection of multiple aflatoxins |
| RU2534732C1 (en) * | 2013-07-26 | 2014-12-10 | Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Национальный исследовательский Томский политехнический университет" | Method for quantitative determination of aflatoxin b1 by differential voltammetry |
| CN105784846A (en) * | 2016-04-08 | 2016-07-20 | 重庆理工大学 | Newcastle disease virus piezoelectric immunosense detection method |
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| CN1168986C (en) * | 2002-01-17 | 2004-09-29 | 大连普瑞康生物技术有限公司 | Aflatoxin fast detecting apparatus and its making method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101216450B (en) * | 2008-01-16 | 2010-09-01 | 暨南大学 | Biosensor electrode for detecting aspergillus flavus toxin B1 and method for making same |
| CN102213687A (en) * | 2010-04-09 | 2011-10-12 | 中国科学院海洋研究所 | Impedance sensor for detecting microbes quickly and construction method thereof |
| CN102692513A (en) * | 2012-06-26 | 2012-09-26 | 长沙理工大学 | Piezoelectric immunosensing chip and device for detecting oncogene C-myc recombinant protein |
| CN103196973A (en) * | 2013-03-13 | 2013-07-10 | 济南大学 | Method for preparing seven-channel aflatoxin immunosensor and application |
| CN103196984A (en) * | 2013-03-13 | 2013-07-10 | 济南大学 | Preparation method and application of sensor for simultaneous detection of multiple aflatoxins |
| CN103196984B (en) * | 2013-03-13 | 2014-04-23 | 济南大学 | Preparation method and application of sensor for simultaneous detection of multiple aflatoxins |
| CN103196973B (en) * | 2013-03-13 | 2014-06-04 | 济南大学 | Method for preparing seven-channel aflatoxin immunosensor and application |
| RU2534732C1 (en) * | 2013-07-26 | 2014-12-10 | Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Национальный исследовательский Томский политехнический университет" | Method for quantitative determination of aflatoxin b1 by differential voltammetry |
| CN105784846A (en) * | 2016-04-08 | 2016-07-20 | 重庆理工大学 | Newcastle disease virus piezoelectric immunosense detection method |
| CN105784846B (en) * | 2016-04-08 | 2018-10-09 | 重庆理工大学 | A kind of newcastle disease virus piezoelectric immuno-sensing detection method |
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