CN107238700A - Quantum dot immune chromatograph test strip and preparation method thereof is quenched in a kind of collaurum for detecting zearalenone - Google Patents
Quantum dot immune chromatograph test strip and preparation method thereof is quenched in a kind of collaurum for detecting zearalenone Download PDFInfo
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- CN107238700A CN107238700A CN201710411662.1A CN201710411662A CN107238700A CN 107238700 A CN107238700 A CN 107238700A CN 201710411662 A CN201710411662 A CN 201710411662A CN 107238700 A CN107238700 A CN 107238700A
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- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
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- 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
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- 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
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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Abstract
Quantum dot immune chromatograph test strip is quenched the invention provides a kind of collaurum for detecting zearalenone, including sample pad 1, nitrocellulose filter 2, adsorptive pads 3 and PVC backboards, characterized in that, being stained with sample pad 1, nitrocellulose filter 2, adsorptive pads 3 successively in order on PVC backboards；The detection line 4 and the nature controlling line 5 of quantum dot chicken ovalbumin conjugate composition of zearalenone antigen and quantum dot chicken ovalbumin conjugate composition are coated with described nitrocellulose filter 3 respectively.The invention also discloses the preparation method of this test strips.Invention has the advantages that following prominent：1st, specificity is high, and sensitivity is good；2nd, testing cost is low.3rd, it is easy to operate.
Description
Technical field
The present invention relates to a kind of method of zearalenone in quick detection cereal foods, particularly a kind of detection corn
Quantum dot immune chromatograph test strip and preparation method thereof is quenched in the collaurum of zeranol.
Background technology
Zearalenone is a kind of mainly by the generation of the strains such as fusarium moniliforme, fusarium tricinctum, Gibberella zeae
The mycotoxin of phenol.Zearalenone has female hormone effect, mainly acts on reproductive system, influence poultry and livestock
Estrogen level, causes domestic animal genital system diseases, is mainly shown as genitals swelling [6], hyperemia, stillborn foetus and miscarriage, spirit
It is tired, anorexia, the problems such as abdominal pain diarrhea, while can also all have certain poison to nervous system, heart, kidney, liver and lung
Evil effect.At present, specific drug there is no to treat the poisoning of animal zearalenone, and general animal zearalenone poisoning
Immediate cause be have in feed go mouldy particularly by corn, wheat, soybean of red mould pollution etc..Therefore, in order to ensure consumption
Person's health must be set up sensitivity height, high specificity, simple and quick assay method and zearalenone is monitored.
Physicochemical property based on zearalenone, ZEN method is detected at present mainly to be included chromatogram analysis method and exempts from
Epidemic disease analysis method.Wherein chromatogram analysis method mainly include thin-layered chromatography (Thin-layer chromatography, TLC),
Gas chromatography (Gas chromatography, GC), high performance liquid chromatography (Liquid chromatography, HPLC)
Deng, immunoassay method mainly include enzyme linked immunosorbent assay (Enzyme linked immunosorbent assay,
ELISA), immunochromatographic method (Immunochromatographic, ICA), chemiluminescence immunoassay
(Chemiluminescence immunoassay, CLIA) etc..Instrument detection method method is to apply most detection sides at present
Method, has the advantages that degree of accuracy height, sensitivity are good.But the shortcoming of this method is, it is necessary to which expensive instrument and the personnel of specialty enter
Row operation, sample handling procedure is cumbersome, and, it is necessary to derivative, the rate of recovery is low, so being not suitable for widely promoting the use of.
Quantum dot (Quantum dots, QDs), also known as semiconductor nano, mainly by II~VI race or III~V
Race's element composition, such as cadmium telluride (CdTe), cadmium selenide (CdSe).Compared with conventional fluorescent material, QDs advantage has：Have
Quantum effect and good photism；With wider excitation wavelength and narrower launch wavelength；Stability is good；With good
Bio-compatibility；Fluorescence lifetime is longer.
It is based on fluorogenic donor (quantum dot) and fluorescent receptor (collaurum) that immune series of strata test strips, which are quenched, in quantum dot fluorescence
Resonance energy transfer (FRET) realize.I.e. when the emission spectrum and fluorescent receptor (collaurum) of fluorogenic donor (quantum dot)
Absorption spectrum has certain overlapping, (is generally less than when the distance between the two fluorophors is suitable), so that it may it was observed that glimmering
The phenomenon that light energy is shifted from donor to acceptor.It is on the basis of immuno analytical method that immunochromatography technique, which is quenched, in quantum dot fluorescence
On, make fluorogenic donor using fluorescence quantum and mixed with envelope antigen, is fixed on NC films, collaurum makees fluorescent receptor and antibody
With reference to when antibody is combined with antigentic specificity, collaurum is just marked on determined antigen, with quantum under the irradiation of uviol lamp
Resonance energy transfer occurs for point.Wherein, the presence or absence of nature controlling line (C lines) color decides the validity of test strips, and detection line (T
Line) the presence or absence of then represent the presence or absence of object.The technical operation is simple and quick, result easily judges, safety non-pollution, has
It is widely applied prospect.
The content of the invention
In view of this, the invention is directed to a kind of detect that the collaurum of zearalenone is quenched in cereal foods
Quantum dot immune chromatograph test strip.
To reach above-mentioned purpose, what the technical scheme of the invention was realized in：
Quantum dot immune chromatograph test strip, including sample pad, nitric acid is quenched in a kind of collaurum for detecting zearalenone
Cellulose membrane, adsorptive pads and PVC backboards, are stained with sample pad, nitrocellulose filter, water suction successively in order on PVC backboards
Pad；Zearalenone antigen and quantum dot-chicken ovalbumin conjugate structure are coated with described nitrocellulose filter respectively
Into the nature controlling line 5 that constitutes of detection line 4 and quantum dot-chicken ovalbumin conjugate, two lines spacing distance is 5mm.
The invention also discloses the preparation method that quantum dot immune chromatograph test strip is quenched in above-mentioned collaurum, including following step
Suddenly：
1. the purifying of zearalenone ascites
Zearalenone ascites used in this experiment is made by oneself by laboratory, is purified using octanoic acid-ammonium sulfate method in ascites
Antibody, concrete operation step is as follows：
1) 1mL ascites is taken, 2mL acetate buffer solutions (0.06mol/L, pH 4.0) is added, 1mol/L is used after mixing
NaOH solution adjusts pH to 4.8.
2) 33 μ L caprylic acids are slowly added in the state of stirring, is positioned at 4 DEG C and stands 2h.
3) at 4 DEG C, 15000rpm centrifugation 30min discard precipitation, collect supernatant.
4) PBS solution (0.1mol/L PBS, pH 7.4) of about supernatant volume 1/10 is added into supernatant, is used
1mol/L NaOH solutions adjust pH to 7.4.
5) ammonium sulfate is slowly added into solution under ice bath state, every milliliter of solution adds 0.277g, stirred after adding
30min。
6) at 4 DEG C, 5000rpm centrifugation 15min discard supernatant, precipitation is molten with PBS (0.01mol/L PBS, pH 7.4)
Solution, 4 DEG C of dialysed overnights.
7) collect at the solution dialysed, 4 DEG C, 5000rpm centrifugation 10min collect supernatant, the body of dilution before purification
Product, i.e. 1mL.
2. the preparation of checking matter antigen (ZEN-OVA)
(1) synthesis of haptens：10mg zearalenones accurately are weighed in round-bottomed flask, and adding 200 Μ l methanol makes
It fully dissolves, and is subsequently added 20mg carboxymethyls azanol (CMO), and 1Ml anhydrous pyridines are stirred at room temperature 24h under argon gas protection, reacted
During with TLC detect the extent of reaction, solvent is dichloromethane：Methanol=9:1(v/v).50 DEG C of vacuum are done after reaction terminates
Dry, 4 DEG C save backup.
(2) preparation of antigen (ZEN-OVA)：Accurately weigh zearalenone hapten conjugation thing 9.78mg and EDC
14mg, then adds 1mLDMF to the inside, activates and stay overnight under conditions of 4 DEG C after stirring.Weigh 10mg OVA and by its
In the sodium bicarbonate buffer liquid for being dissolved in 2mL (130mmol/L, pH=8.1), precooling, and activation is added dropwise under condition of ice bath
Good product, reacts under the conditions of 4 DEG C and stays overnight again after reaction 2h under conditions of room temperature.Products therefrom is dialysed with PBS solution
72h, determines concentration and is dispensed and preserved at -20 DEG C according to dosage different every time.
3. for the preparation of quantum dot-chicken ovalbumin conjugate：
Quantum dot-chicken ovalbumin (QDs-OVA) conjugate is prepared using active ester method.
(1) coupling reaction.50 μ L quantum dots are taken in centrifuge tube, 0.6mg chicken ovalbumins and 23 μ L EDC solutions are added
(10mg/ml), is mixed, and liquor capacity is supplemented into 400 μ L with borate buffer solution.Then with aluminium-foil paper parcel peace deferent,
Reaction 3h is shaked in shaking table lucifuge.
(2) centrifugal purification.By reaction solution under the conditions of 4 DEG C, 10000rpm centrifugation 3min are formed with removing in labeling process
Coagulation thing；Supernatant is transferred in filter membrane, under the conditions of 4 DEG C, 8000rpm centrifugation 3min remove the salt ion in solution；
Finally filter membrane is upside down in another centrifuge tube, under the conditions of 4 DEG C, 8000rpm centrifugation 3min collect the solution in filter membrane.4℃
It is kept in dark place standby.
4. the preparation of collaurum-zearalenone antibody (AuNPs-ZEN-Ab) label
(1) the accurate 1mL colloidal gold solutions that pipette are placed in import peace deferent, add 5 μ L 0.2mol/L K2CO3Solution, gently
Shake peace deferent to mix, regulation pH value to optimum value.
(2) 5 μ L 1mg/mL zearalenone antibody (ZEN-Ab) is added in above-mentioned solution and be well mixed, put
Put the 1h in 4 DEG C of refrigerators.
(3) add thereto 20 μ L concentration be 20% BSA solution to stablize gold labeling antibody, add 10 μ L's 20%
The solution of PEG 20000, other sites that closing gold colloid surface is not connected with antibody, stands 30min at room temperature.
(4) by peace deferent trim, 15min is centrifuged under the conditions of 4 DEG C of 2000rpm, the gold particle for being not connected with upper antibody is reunited, shape
Into precipitation, Aspirate supernatant is transferred in another peace deferent.
(5) again with 4 DEG C, 10000rpm pelleted by centrifugation 30min, at this moment gold labeling antibody formation precipitation.
(6) supernatant is siphoned away, precipitation is preserved, is resuspended with gold mark working solution.
5. the coating of nitrocellulose filter
With the double dimensional planes of the golden target in Shanghai draw ZEN-OVA that film instrument prepare step 3 PBS solution dilute after 3 times with use
The QDs-OVA labels that PBS solution is diluted after 6 times are coated on nitrocellulose filter 3 as detection line 5, and package amount is 0.75 μ
L/cm；It is coated in the QDs-OVA after 12 times is diluted with PBS solution as nature controlling line 6 on nitrocellulose filter 3, package amount is
0.75 μ L/cm, 37 DEG C of drying are encapsulated standby.
6. the AuNPs-ZEN-Ab labels for taking 8 μ L steps 4 to prepare, make an addition in sample solution；
7. adhering to sample pad, nitrocellulose filter, adsorptive pads successively in order on PVC backboards, described corn is obtained
Quantum dot immune chromatograph test strip is quenched in zeranol collaurum.
In above-mentioned material, water-soluble quantum dot is purchased from Wuhan Jia Yuan technology of quantum dots development corporation, Ltd., sample pad and nitre
Acid cellulose film is purchased from Millipore companies of the U.S., and adsorptive pads and PVC backboards are purchased from Shanghai Jin Biao biotechnologies company.
The present invention has purified checking matter ascites, has obtained checking matter monoclonal antibody, standby.Prepare ZEN-OVA mixing
QDs-OVA labels are the envelope antigen of detection line, and nature controlling line is marked using QDs-OVA.Detect that corn is red using competition law
Mould ketenes, judges whether contain object in testing sample according to the depth of detection line band.If nature controlling line band is without color,
The test strips fail.
Compared with the method for existing domestic and international detection zearalenone, the present invention has the advantages that following prominent：1st, originally
Quantum dot immune chromatograph test strip, which is quenched, in the collaurum of invention is realized using the specific reaction of antigen-antibody, therefore specifically
Property it is high, sensitivity is good.2nd, test strip detection time of the invention quick (15 minutes) and do not need any specific apparatus and
Equipment, testing cost is low.3rd, test strip of the invention is easy to operate, is not required to be operated by professional.
Brief description of the drawings
The accompanying drawing for constituting the part of the invention is used for providing further understanding the invention, present invention wound
The schematic description and description made is used to explain the invention, does not constitute the improper restriction to the invention.
In accompanying drawing：
Fig. 1 is the assembling schematic diagram of test strip of the present invention.
Fig. 2 is the result judgement method schematic diagram of test strip of the present invention.
Fig. 3 for zearalenone test limit determination (concentration of zearalenone from left to right be respectively 0,0.25,
0.5、1、2、5μg/L)。
Fig. 4 is the food samples testing result of addition zearalenone in corn sample.Every group of zearalenone
Concentration is respectively from left to right 0,1.5,3,6,12,30 μ g/kg.
Fig. 5 is the food samples testing result of addition zearalenone in wheat samples.Every group of zearalenone
Concentration is respectively from left to right 0,1.5,3,6,12,30 μ g/kg.
Description of reference numerals：
1st, sample pad, 2, nitrocellulose filter 3, adsorptive pads, 4, detection line,
5th, nature controlling line, 6, PVC board
Embodiment
It should be noted that in the case where not conflicting, the feature in embodiment and embodiment in the invention can
To be mutually combined.
Describe the invention in detail below with reference to the accompanying drawings and in conjunction with the embodiments.
Embodiment l (preparation embodiment)
(1) purifying of zearalenone ascites
Zearalenone ascites used in this experiment is made by oneself by laboratory, is purified using octanoic acid-ammonium sulfate method in ascites
Antibody, concrete operation step is as follows：
1) 1mL ascites is taken, 2mL acetate buffer solutions (0.06mol/L, pH 4.0) is added, 1mol/L is used after mixing
NaOH solution adjusts pH to 4.8.
2) 33 μ L caprylic acids are slowly added in the state of stirring, is positioned at 4 DEG C and stands 2h.
3) at 4 DEG C, 15000rpm centrifugation 30min discard precipitation, collect supernatant.
4) PBS solution (0.1mol/L PBS, pH 7.4) of about supernatant volume 1/10 is added into supernatant, is used
1mol/L NaOH solutions adjust pH to 7.4.
5) ammonium sulfate is slowly added into solution under ice bath state, every milliliter of solution adds 0.277g, stirred after adding
30min。
6) at 4 DEG C, 5000rpm centrifugation 15min discard supernatant, precipitation is molten with PBS (0.01mol/L PBS, pH 7.4)
Solution, 4 DEG C of dialysed overnights.
7) collect at the solution dialysed, 4 DEG C, 5000rpm centrifugation 10min collect supernatant, the body of dilution before purification
Product, i.e. 1mL.
(2) synthesis of zearalenone haptens
10mg zearalenones accurately are weighed in round-bottomed flask, and adding 200 Μ l methanol makes it fully dissolve, then
20mg carboxymethyls azanol (CMO) is added, 1Ml anhydrous pyridines are stirred at room temperature in 24h, course of reaction under argon gas protection and detected with TLC
The extent of reaction, solvent is dichloromethane：Methanol=9:1(v/v).50 DEG C of vacuum drying after reaction terminates, 4 DEG C save backup.
(3) preparation of zearalenone antigen (ZEN-OVA)
(1) zearalenone hapten conjugation thing 9.78mg and EDC 14mg accurately is weighed, is then added to the inside
1mLDMF, activates under conditions of 4 DEG C after stirring and stays overnight.Weigh 10mg OVA and be dissolved in 2mL (130mmol/L, pH
In=sodium bicarbonate buffer liquid 8.1), precooling, and the product activated is added dropwise under condition of ice bath, in the condition of room temperature
React and stay overnight under the conditions of 4 DEG C again after lower reaction 2h.By products therefrom PBS solution dialysis 72h, concentration is determined and according to each
Different dosage is dispensed and preserved at -20 DEG C.
Mixed using the antigen with quantum dot-chicken ovalbumin QDs-OVA, the detection line on coating nitrocellulose filter.
Embodiment 2 (preparation embodiment)
Assembling and the preparation method of immuno-chromatographic test paper strip is quenched in quantum dot fluorescence
1st, test strips are assembled：
The test strips composition of the present invention is as follows：Sample pad 1, nitrocellulose filter 2, adsorptive pads 3 and PVC board 6, in PVC board 6
On be stained with sample pad 1, nitrocellulose filter 2, adsorptive pads 3 successively in order；It is coated with respectively on described nitrocellulose filter 2
There are the detection line 4 that zearalenone antigen is constituted and the nature controlling line 5 that goat-anti rabbit secondary antibody is constituted.
2nd, the preparation of standby quantum dot-chicken ovalbumin conjugate：
Quantum dot-chicken ovalbumin (QDs-OVA) conjugate is prepared using active ester method.
(1) coupling reaction.50 μ L quantum dots are taken in centrifuge tube, 0.6mg chicken ovalbumins and 23 μ L EDC solutions are added
(10mg/ml), is mixed, and liquor capacity is supplemented into 400 μ L with borate buffer solution.Then with aluminium-foil paper parcel peace deferent,
Reaction 3h is shaked in shaking table lucifuge.
(2) centrifugal purification.By reaction solution under the conditions of 4 DEG C, 10000rpm centrifugation 3min are formed with removing in labeling process
Coagulation thing；Supernatant is transferred in filter membrane, under the conditions of 4 DEG C, 8000rpm centrifugation 3min remove the salt ion in solution；
Finally filter membrane is upside down in another centrifuge tube, under the conditions of 4 DEG C, 8000rpm centrifugation 3min collect 4 DEG C of solution in filter membrane
Save backup.
The preparation of 3 collaurums-zearalenone antibody (AuNPs-ZEN-Ab) label
(1) the accurate 1mL colloidal gold solutions that pipette are placed in import peace deferent, add 5 μ L 0.2mol/L K2CO3Solution, gently
Shake peace deferent to mix, regulation pH value to optimum value.
(2) 5 μ L 1mg/mL zearalenone antibody (ZEN-Ab) is added in above-mentioned solution and be well mixed, put
Put the 1h in 4 DEG C of refrigerators.
(3) add thereto 20 μ L concentration be 20% BSA solution to stablize gold labeling antibody, add 10 μ L's 20%
The solution of PEG 20000, other sites that closing gold colloid surface is not connected with antibody, stands 30min at room temperature.
(4) by peace deferent trim, 15min is centrifuged under the conditions of 4 DEG C of 2000rpm, the gold particle for being not connected with upper antibody is reunited, shape
Into precipitation, Aspirate supernatant is transferred in another peace deferent.
(5) again with 4 DEG C, 10000rpm pelleted by centrifugation 30min, at this moment gold labeling antibody formation precipitation.
(6) supernatant is siphoned away, precipitation is preserved, is resuspended with gold mark working solution.
The coating of 4 nitrocellulose filters
The zearalenone antigen after 3 times will be diluted with PBS solution with using by drawing film instrument with the Shanghai double dimensional planes of golden target
PBS solution dilutes the QDs-OVA after 6 times, and mixing is coated in as detection line 5 on nitrocellulose filter 3 in equal volume, and package amount is
0.75μL/cm；The QDs-OVA that will be diluted with PBS solution after 12 times, which is coated on nitrocellulose filter 3, is used as nature controlling line 6, is coated with
Measure as 0.75 μ L/cm, 37 DEG C of drying are encapsulated standby.
The assembling of 5 test strips
Sample pad 1, nitrocellulose filter 2, the order of adsorptive pads 3 as shown in Figure 1 are sticked in PVC board 6 successively, are cut into
Small bar wide 3.7mm, Vacuum Package.
Embodiment 3 (Application Example)
1st, quantum dot-labeled immunochromatographyassay assay test-strip application method
The pretreatment of grain sample
5g grain samples are placed in after grinding in 50mL centrifuge tubes, add 5mL 70% methanol solution, 5min is mixed at a high speed,
Volume is supplemented to 30mL under the conditions of 4 DEG C with phosphate buffer (pH 7.5), 5000g centrifugation 5min, Aspirate supernatant, directly
Connect for ELISA test strip.
2nd, detecting step
200 μ L measuring samples extract solutions are drawn in peace deferent with liquid-transfering gun, and add 8 μ L AuNPs-ZEN-Ab, are mixed
Afterwards, it is added drop-wise in the well of test strips, result is observed after 15 minutes.
3rd, result judgement
If as shown in Fig. 2 testing sample ELISA test strip line fluorescence disappears, being judged as negative sample, i.e., without corn
Zeranol；If testing sample ELISA test strip line fluorescence be shallower than Quality Control line color or with nature controlling line solid colour, be judged as
Contain zearalenone in positive, i.e. testing sample；Positive findings and negative findings, nature controlling line show green fluorescence bar
Band, if nature controlling line green fluorescence band disappears, ELISA test strip failure.
Embodiment 4 (Application Example)
The application effect citing of the present invention
Signified protein anabolic hormone quantum dot-labeled immunochromatographyassay assay test-strip detection method is with reference to implementation in the present embodiment
Operating procedure described in example 3, its testing result is as follows.
1st, sensitivity test
As shown in figure 3, being tested by the methods described of embodiment 3.When zearalenone standard concentration is less than 0.25 μ
During g/L, ELISA test strip line unstressed configuration band, colour developing result is compared with the range estimation of Quality Control line color significant difference；When corn is red
When mould ketenes standard concentration is 0.25 μ g/L, there is fluorescent bands, colour developing result and Quality Control line color mesh in ELISA test strip line
It is smaller that difference is compared in survey；When zearalenone standard concentration continues to increase, detection line gradually brightens；Zearalenone mark
When quasi- product concentration is 0.5 μ g/L, detection line colour developing result is consistent with the range estimation of Quality Control line color.It is thus determined that the visual detection of method
It is limited to 0.25 μ g/L.
2nd, the detection of mark-on sample
As shown in figure 4, it is 0,1.5,3 that ultimate density in zearalenone standard items, sample is added into corn sample,
6,12, and 30 μ g/kg, PBS solution added body is used after 70% methanol of 5mL-PBS solution, concussion 5min are added into 5g samples
Product is to 30mL, and 5000rpm is centrifuged after 5min after mixing, takes supernatant to be detected.Tested by the methods described of embodiment 3,
Testing result such as Fig. 4.
As shown in figure 5, it is 0,1.5,3 that ultimate density in zearalenone standard items, sample is added into wheat samples,
6,12, and 30 μ g/kg, PBS solution added body is used after 70% methanol of 5mL-PBS solution, concussion 5min are added into 5g samples
Product is to 30mL, and 5000rpm is centrifuged after 5min after mixing, takes supernatant to be detected.Tested by the methods described of embodiment 3,
Testing result such as Fig. 5.
Experiment show the present invention test strips accuracy is good, sensitivity is high, and sample-pretreating method is simple, whole inspection
Survey process is no more than 20min, it is adaptable to the quick screening of a large amount of samples, can be as being to remain quick detection protein anabolic hormone more
Effective screening means.
The preferred embodiment of the invention is the foregoing is only, creation is not intended to limit the invention, it is all at this
Within the spirit and principle of innovation and creation, any modification, equivalent substitution and improvements made etc. should be included in the invention
Protection domain within.
Claims (3)
1. quantum dot immune chromatograph test strip, including sample pad, nitric acid fibre is quenched in a kind of collaurum for detecting zearalenone
The plain film of dimension, adsorptive pads and PVC backboards, it is characterised in that be stained with sample pad, cellulose nitrate successively in order on PVC backboards
Plain film, adsorptive pads；Zearalenone antigen and quantum dot-chicken ovalbumin are coated with described nitrocellulose filter respectively
The nature controlling line e that the detection line d and quantum dot-chicken ovalbumin conjugate that conjugate is constituted are constituted, two lines spacing distance is
5mm。
2. quantum dot immune chromatographic test paper is quenched in a kind of collaurum for detecting zearalenone according to claim 1
Bar, it is characterised in that for detecting zearalenone.
3. the system of quantum dot immune chromatograph test strip is quenched in a kind of collaurum of detection zearalenone described in claim 1
Preparation Method, it is characterised in that comprise the following steps：
(1) zearalenone ascites is purified, and obtains checking matter monoclonal antibody；
(2) checking matter antigen is prepared；
(3) quantum dot-chicken ovalbumin conjugate is prepared using active ester method；
(4) collaurum-antibody labeling thing is prepared；
(5) the checking matter collaurum-antibody labeling thing for taking appropriate step (4) to prepare, makes an addition in sample solution；
(6) checking matter for taking checking matter quantum dot-chicken ovalbumin label prepared by appropriate step (3) to be prepared with step (2)
Antigen mixing, which is coated on nitrocellulose filter, constitutes detection line, and the checking matter quantum dot-ovum gallinaceum for taking appropriate step (3) to prepare is white
Protein marker, which is coated on nitrocellulose filter, constitutes nature controlling line；
(7) sample pad, nitrocellulose filter, adsorptive pads are adhered to successively in order on PVC backboards, described Gibberella zeae is obtained
Quantum dot immune chromatograph test strip is quenched in ketenes collaurum.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109061144A (en) * | 2018-09-19 | 2018-12-21 | 无限极（中国）有限公司 | A kind of qualitative, quantitative immuno-chromatographic test paper strip and its preparation method and application for detecting zearalenone |
| CN109900890A (en) * | 2019-03-28 | 2019-06-18 | 天津科技大学 | A kind of black phosphorus-gold nanoparticle compound photo-thermal quantitative immunochromatographic test strips and preparation method thereof detecting small-molecule substance |
| CN110330974A (en) * | 2019-07-11 | 2019-10-15 | 南京工业大学 | Preparation and application of zearalenone ratiometric fluorescent probe |
| CN110396126A (en) * | 2019-07-23 | 2019-11-01 | 山东绿都生物科技有限公司 | A kind of monoclonal antibody and its application identifying zearalenone |
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2017
- 2017-06-05 CN CN201710411662.1A patent/CN107238700A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109061144A (en) * | 2018-09-19 | 2018-12-21 | 无限极（中国）有限公司 | A kind of qualitative, quantitative immuno-chromatographic test paper strip and its preparation method and application for detecting zearalenone |
| CN109900890A (en) * | 2019-03-28 | 2019-06-18 | 天津科技大学 | A kind of black phosphorus-gold nanoparticle compound photo-thermal quantitative immunochromatographic test strips and preparation method thereof detecting small-molecule substance |
| CN110330974A (en) * | 2019-07-11 | 2019-10-15 | 南京工业大学 | Preparation and application of zearalenone ratiometric fluorescent probe |
| CN110330974B (en) * | 2019-07-11 | 2022-09-09 | 南京工业大学 | Preparation and application of zearalenone ratiometric fluorescent probe |
| CN110396126A (en) * | 2019-07-23 | 2019-11-01 | 山东绿都生物科技有限公司 | A kind of monoclonal antibody and its application identifying zearalenone |
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