CN110220891A - A kind of portable mycotoxin immunochromatography quantitative testing device and detection method - Google Patents
A kind of portable mycotoxin immunochromatography quantitative testing device and detection method Download PDFInfo
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- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
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Abstract
The invention discloses a kind of portable mycotoxin immunochromatography quantitative testing device and detection methods.The portable mycotoxin immunochromatography quantitative testing device includes mycotoxin immunochromatography quantitative test card；The mycotoxin immunochromatography quantitative test card includes bottom plate, and sample pad, bonding pad, nitrocellulose filter and blotting paper are successively arranged on bottom plate；Blotting paper and bonding pad overlap the both ends for being pressed in nitrocellulose filter respectively, form detection zone on the surface of nitrocellulose filter；Sample pad is overlapping to be pressed on bonding pad；The mycotoxin antibody of colloidal gold, color latex microballoon or fluorescent microsphere label is fixed on bonding pad；The nature controlling line that the detection line and sheep anti mouse or goat anti-rabbit igg secondary antibody that mycotoxin antigen composition is fixed on nitrocellulose filter in detection zone are constituted.Device is able to achieve to the cereal such as corn and soybean, wheat and feed equal samples scene fast and accurately quantitative detection through the invention, and can be realized the quick transmission of data.
Description
Technical field
The present invention relates to technical field of immunoassay, and in particular, to a kind of portable mycotoxin immunochromatography is quantitative
Detection device and detection method.
Background technique
Mycotoxin (Mycotoxin) is the toxic secondary metabolite for being generated, being widely present in agricultural product by fungi,
Mainly there are aflatoxin (Aflatoxin, AFT), ochratoxin (Ochratoxin, OTA), zearalenone
(Zearalenone, ZEN) etc. can pollute the food and feed of almost all kinds.It is counted according to FAO (Food and Agriculture Organization of the United Nation) (FAO),
The global agricultural product for having 25% every year are polluted by mycotoxin, caused by economic loss reach hundreds billion of dollars every year.These are mould
The food or feed link of change are crossed after food chain enters human body, and carcinogenic, teratogenesis, mutagenesis etc. can be caused to seriously endanger.Mycotoxin
Pollution not only causes huge economic loss, also becomes the material risk hidden danger for threatening human health safety.Therefore, reinforce true
The prevention and control of verticillium toxin have become the urgent need for ensureing China or even world food safety, food safety and Environmental security.
The residue detection of mycotoxin mainly has instrumental method and immunoassay.Instrumental method high sensitivity, specifically
Property it is strong, but need expensive equipment, professional operator and cumbersome sample pre-treatments, be not suitable for the quick sieve of great amount of samples
It looks into.And immunoassay method is quick and easy with its, at low cost, high sensitivity, is suitble in actual production on-site supervision and extensive
The advantages such as sample screening are quickly grown in mycotoxin detection field.It is ELISA that immuno analytical method is most widely used at present
And immune chromatography method.Immune chromatography method because Sample pretreatment is simple, result can be due to 5~20min be obtained by researcher
With the favor of user.But most of immunochromatography detection method can only realize qualitative or half-quantitative detection, while existing quantitative
That there are volumes is big for most of device or system of detection, data processing is inconvenient, accuracy is low, cannot issue result or inspection in time
It surveys result and is easy a series of problems such as error.As used colorimetric card to realize immunochromatography skill in patent CN201520397285.7
The half-quantitative detection of art, because of the influence of the objective condition such as the examined temperature of immuno-chromatographic assay technology, humidity, colored intensity has
Biggish variation, the accurate quantitative analysis of the quantitative approach of colorimetric card colorimetric immunochromatography technique relatively difficult to achieve；Patent
CN201510415935.0 discloses a kind of quantitative measurement technology that immunochromatography technique is realized by photosensitive sensors, at data
It manages cumbersome and can not quickly transmission sample testing result and geography information；Jane RuChoi and Martina Zangheri research and development
The immunochromatography quantitative testing device based on smart phone can preferably realize quantitative detection, but detection device volume it is larger or
The fogging image of acquisition and only show that standard curve is unfavorable for accurately realizing a large amount of samples by one group of standard sample data
The detection of this in situ quantitation (Jane RuChoi et al., 2016；Martina Zangheri et al.,2015).For above-mentioned
Problem is not yet put forward effective solutions in the prior art.It is fixed therefore, it is necessary to research and develop a kind of new mycotoxin immunochromatography
Amount detecting device is to solve the above problems.
Summary of the invention
The purpose of the invention is to overcome the above-mentioned deficiency of the prior art, a kind of immune layer of portable mycotoxin is provided
Quantitative testing device and detection method are analysed, device is able to achieve to samples such as the cereal such as corn and soybean, wheat and feeds through the invention
This scene fast and accurately quantitative detection, and can be realized the quick transmission of data.
To achieve the goals above, the present invention is achieved by following scheme:
A kind of mycotoxin immunochromatography quantitative test card, including bottom plate are successively arranged sample pad on the bottom plate, combine
Pad, nitrocellulose filter and blotting paper；The blotting paper and bonding pad overlap the both ends for being pressed in nitrocellulose filter respectively, in nitre
The surface of acid cellulose film forms detection zone；The sample pad is overlapping to be pressed on bonding pad；Colloid is fixed on the bonding pad
The mycotoxin antibody of gold, color latex microballoon or fluorescent microsphere label；It is fixed on nitrocellulose filter in the detection zone
The nature controlling line (C line) that the detection line (T line) and sheep anti mouse or goat anti-rabbit igg secondary antibody for having mycotoxin antigen to constitute are constituted.
Preferably, the detection line is located at one end in detection zone close to bonding pad, and the nature controlling line is located in detection zone
Close to one end of blotting paper.
Preferably, the mycotoxin immunochromatography quantitative test card further includes shell, the bottom plate, sample pad, combination
Pad, nitrocellulose filter and blotting paper are placed in shell, and the position in the shell shell surface corresponding to nitrocellulose membrane is equipped with
Observation window, the position that sample pad is corresponded in shell shell surface are equipped with well.
Preferably, the sample pad is dry after the buffer immersion treatment containing surfactant, protein protective agent
Glass fibre component.
Preferably, the bottom plate is polystyrene component or polyethylene component.
Preferably, the shell includes plastics upper casing and plastics lower casing, and the plastics upper casing is formed after fastening plastics lower casing
Shell.
A kind of portable mycotoxin immunochromatography quantitative testing device, including above-mentioned fungi poison is also claimed in the present invention
Plain immunochromatography quantitative test card, sequentially connected camera bellows, micro-lens, handset bracket and smart phone, the mycotoxin
Immunochromatography quantitative test card is embedded in camera bellows.
The present invention can be separately designed according to the size of mycotoxin immunochromatography quantitative test card and smart phone camera bellows and
The specification of handset bracket passes through smart phone in the camera bellows of mycotoxin immunochromatography quantitative test card insertion dimension
The light path system of rear camera and micro-lens composition realizes the acquisition of image；By the self-programming for being built in smart phone
Quantitative APP inspection software, is handled the rgb signal of rear camera captured image and is converted to corresponding numerical value, is led to
Standard curve is established in a series of detection for crossing concentration, realizes the detection and analysis to sample to be tested.
Specific testing principle is as follows: in sample concentration of mycotoxins constantly increases immunochromatography system (i.e. very
Verticillium toxin immunochromatography quantitative test card) in T line (detection line) colour developing it is more shallow, (be sequentially connected by image capturing system
Camera bellows, micro-lens, handset bracket and smart phone) a series of concentration samples of acquisition image information, pass through image analysis
In artificial mode reading immunity-chromatography detection card in system (self-programming for being built in smart phone quantifies APP inspection software)
TGray valueAnd CGray valueAnd T is calculated automaticallyGray value/CGray value, it is cross with concentration of mycotoxins by analyzing a series of sample information of concentration
Coordinate, TGray value/CGray valueFor ordinate fit standard curve, and the T for passing through unknown sampleGray value/CGray valueObtain unknown sample concentration
Or the data of Cloud Server are established by machine learning on computers by rule by AI mode, compare the related letter of position sample
Breath can obtain the concentration of unknown sample.
Wherein, on the one hand the camera bellows plays the role of fixed mycotoxin detection card, on the other hand mentions for Image Acquisition
For dark surrounds, it is allowed to free from the influence of the external environment.
Preferably, the camera bellows be photosensitive resin made of 3D printing.
The micro-lens can shorten shooting distance, keep whole device more portable；Captured area can be amplified again
Domain is more clear shooting area, is conducive to data acquisition, reads and analyze.
Preferably, the micro-lens includes the first micro-lens and the second micro-lens being connected, the first Macro Lens
The other end of head connects with the rear camera of smart phone, and the other end of the second micro-lens connects with camera bellows.Pass through setting
Two micro-lens make single unit system more compact, portable and can collect the image being more clear.
The handset bracket is connect with smart phone, and whole device is fixed on smart phone.
The smart phone reads image information, built-in quantitative APP inspection software point by rear camera and flash lamp
Analyse image information.
The quantitative APP inspection software includes project management plate, can create difference for different types of mycotoxin
Project.Image Acquisition plate acquires pictorial information, and can be uploaded to Cloud Server；Image analysis plate, is divided into artificial mode
With AI mode, artificial mode calculates T by the RGB information of one group of master sample image of analysisGray value/CGray value, and fit standard is bent
Line, realizes quantitative detection, and AI pattern analysis is uploaded to the great amount of samples information on backstage；Testing result module shows testing result
And it is geographical to network to position upload.
A kind of detection method of mycotoxin is also claimed in the present invention, is that layer is immunized using above-mentioned portable mycotoxin
Analysis quantitative testing device detects mycotoxin standard sample or sample to be tested；First pass through smart phone rear camera with
And the light path system of micro-lens composition realizes the acquisition of image；APP inspection is quantified by being built in the self-programming of smart phone again
Software is surveyed, the rgb signal of rear camera captured image is handled and is converted to corresponding numerical value；Finally by a system
Standard curve is established in the detection of column concentration, realizes the quantitative detecting analysis to mycotoxin in sample to be tested.
Preferably, the quantitative APP inspection software includes two kinds of operational modes of artificial mode and AI mode；When sample size is small
Shi Caiyong artificial mode calculates T by the RGB information of one group of master sample image of analysisGray value/CGray value, and fit standard curve,
Realize quantitative detection；AI mode is used when sample size is big, and the data of Cloud Server are passed through by machine by AI mode on computers
Rule is established in device study, and the relevant information for comparing position sample obtains the concentration of unknown sample.
Preferably, the detection method includes the following steps:
S1. the mycotoxin standard of equivalent various concentration is added into mycotoxin immunochromatography quantitative test card well
Sample reacts 3~5min；
S2. operation mycotoxin immunochromatography quantitative detection APP Image Acquisition plate acquires image and is uploaded to cloud service
Device；
S3. operation mycotoxin immunochromatography quantitative detection APP image analysis plate artificial mode obtains sample RGB information
With examination criteria curve；If sample size is big, using AI mode；
S4. appropriate mycotoxin sample to be measured, reaction 3 are added into mycotoxin immunochromatography quantitative test card well
~5min；
S5. operation mycotoxin immunochromatography quantitative detection APP Image Acquisition plate acquires image and is uploaded to cloud service
Device；
S6. by operation mycotoxin immunochromatography quantitative detection APP testing result plate color developing detection as a result, and uploading
As a result with sample geography information to network.
Preferably, the mycotoxin is zearalenone.
Preferably, in the mycotoxin immunochromatographydetection detection card, blue latex microballoon is fixed on the bonding pad
The zearalenone monoclonal antibody of label；It is anti-that zearalenone is fixed on nitrocellulose filter in the detection zone
The nature controlling line that the detection line and sheep anti-mouse igg secondary antibody that former ZEN-OVA is constituted are constituted.
Compared with prior art, the invention has the following advantages:
(1) portable mycotoxin immunochromatography quantitative testing device provided by the present invention has small in size, operation letter
Just, fast, the accurate advantage of detection time, and qualitative detection can be carried out, can quickly will test result by wireless network into
Row is shared.Not only it had been able to achieve to the cereal such as corn and soybean, wheat and feed equal samples scene fast and accurately quantitative detection, but also energy
Enough realize the quick transmission of data.
(2) the smart phone detection system being currently known, primarily directed to signals such as liquid RGB, fluorescence, and it is of the invention
Device is to realize the quick of mycotoxin, accurate quantitative analysis detection for solid-phase immunity chromatograph test strip rgb signal in a creative way.
(3) it is compared with known same type portable detector, the portable mycotoxin immunochromatography of the present invention is quantitatively examined
Device is surveyed in terms of detecting APP, increases AI detection pattern on the basis of the detection module of same type detection device, by sample message
It is uploaded to Cloud Server, batch processing is carried out to sample message by machine learning rule from the background, to be examined in batch samples
Greatly reduce detection time in survey, and go out in data processing in can preventing from detecting at the scene since testing staff is not professional
The problems such as existing mistake or artificial modification data.
Detailed description of the invention
Fig. 1 is the overall structure diagram of mycotoxin immunochromatography quantitative test card.
Fig. 2 is the overall structure diagram of portable mycotoxin immunochromatography quantitative testing device.
Fig. 3 is the side view of portable mycotoxin immunochromatography quantitative testing device.
Fig. 4 is that the overall structure of portable mycotoxin immunochromatography quantitative testing device splits schematic diagram.
Fig. 5 is that the overall structure of portable mycotoxin immunochromatography quantitative testing device splits side view.
Appended drawing reference: 1- mycotoxin immunochromatography quantitative test card；2- camera bellows；3- micro-lens；4- handset bracket；5-
Smart phone；11- bottom plate；12- sample pad；13- bonding pad；14- nitrocellulose filter；15- blotting paper；16- shell；17- is seen
Examine window；18- well；The first micro-lens of 31-；The second micro-lens of 32-.
Specific embodiment
With reference to the accompanying drawings of the specification and specific embodiment is made the present invention and is further elaborated, the embodiment
It is served only for explaining the present invention, be not intended to limit the scope of the present invention.Test method as used in the following examples is such as without spy
Different explanation, is conventional method；Used material, reagent etc., unless otherwise specified, for the reagent commercially obtained
And material.
Embodiment 1
A kind of mycotoxin immunochromatography quantitative test card 1, as shown in Figure 1, including shell 16 and bottom plate 11, on bottom plate 11
It is successively arranged sample pad 12, bonding pad 13, nitrocellulose filter 14 and blotting paper 15；Blotting paper 15 and bonding pad 13 overlap respectively
The both ends of nitrocellulose filter 14 are pressed in, form detection zone on the surface of nitrocellulose filter 14；Sample pad 12 is overlapping to be pressed in knot
It closes on pad 13；The mycotoxin antibody of colloidal gold, color latex microballoon or fluorescent microsphere label is fixed on bonding pad 13；Detection
The detection line and sheep anti mouse or goat anti-rabbit igg secondary antibody structure that mycotoxin antigen is constituted are fixed on nitrocellulose filter 14 in area
At nature controlling line；Detection line is located at one end in detection zone close to bonding pad 13, and nature controlling line is located in detection zone close to blotting paper
15 one end；Bottom plate 11, sample pad 12, bonding pad 13, nitrocellulose filter 14 and blotting paper 15 are placed in shell 16, shell
It is equipped with observation window 17 corresponding to the position of nitrocellulose membrane 14 in 16 shell surfaces, the position of sample pad 12 is corresponded in 16 shell surface of shell
Equipped with well 18；Shell 16 includes plastics upper casing and plastics lower casing, and plastics upper casing forms shell 16 after fastening plastics lower casing.
Wherein, sample pad 12 is dry glass after the buffer immersion treatment containing surfactant, protein protective agent
Fibrous structures；Bottom plate 11 is polystyrene component or polyethylene component.
Embodiment 2
A kind of portable mycotoxin immunochromatography quantitative testing device, as shown in Figure 2 to 4, including in embodiment 1
Mycotoxin immunochromatography quantitative test card 1, sequentially connected camera bellows 2, micro-lens 3, handset bracket 4 and smart phone 5, very
Verticillium toxin immunochromatography quantitative test card 1 is embedded in camera bellows 2；Camera bellows 2 be photosensitive resin made of 3D printing；Microspur
Camera lens 3 includes the first micro-lens 31 and the second micro-lens 32 being connected, the other end and intelligence of the first micro-lens 31
The rear camera of mobile phone 5 connects, and the other end of the second micro-lens 32 connects with camera bellows 2.
Wherein, 2 one side of camera bellows plays the role of fixed mycotoxin detection card 1, on the other hand provides for Image Acquisition
Dark surrounds is allowed to free from the influence of the external environment.Micro-lens 3 can shorten shooting distance, make whole device more just
It takes；Captured region can be amplified again, be more clear shooting area, be conducive to data acquisition, read and analyze.Pass through setting two
A micro-lens makes single unit system more compact, portable and can collect the image being more clear.Handset bracket 4 and intelligence
Mobile phone 5 connects, and whole device is fixed on smart phone 5.Smart phone 5 reads image by rear camera and flash lamp
Information, built-in quantitative APP inspection software analyze image information.Quantitative APP inspection software includes project management plate, for difference
The mycotoxin of type can create different projects.Image Acquisition plate acquires pictorial information, and can be uploaded to Cloud Server；
Image analysis plate is divided into artificial mode and AI mode, the RGB information meter that artificial mode passes through one group of master sample image of analysis
Calculate TGray value/CGray value, and fit standard curve, realize quantitative detection, AI pattern analysis is uploaded to the great amount of samples information on backstage；Inspection
Object module is surveyed, show testing result and upload can be positioned is geographical to network.
Camera bellows 2 and mobile phone can be separately designed according to the size of mycotoxin immunochromatography quantitative test card 1 and smart phone 5
Mycotoxin immunochromatography quantitative test card 1 is inserted into the camera bellows 2 of dimension, passes through smart phone 5 by the specification of bracket 4
The light path system of rear camera and micro-lens composition realizes the acquisition of image；By being built in the self-editing of smart phone 5
Cheng Dingliang APP inspection software is handled the rgb signal of rear camera captured image and is converted to corresponding numerical value,
By a series of detection of concentration, standard curve is established, realizes the detection and analysis to sample to be tested.
Specific testing principle is as follows: in sample concentration of mycotoxins constantly increases immunochromatography system (i.e. very
Verticillium toxin immunochromatography quantitative test card 1) in T line (detection line) colour developing it is more shallow, (be sequentially connected by image capturing system
Camera bellows 2, micro-lens 3, handset bracket 4 and smart phone 5) a series of concentration samples of acquisition image information, pass through image
Artificial mode reading immunity-chromatography inspection in analysis system (self-programming for being built in smart phone 5 quantifies APP inspection software)
Survey T in card 1Gray valueAnd CGray valueAnd T is calculated automaticallyGray value/CGray value, by analyzing a series of sample information of concentration, with mycotoxin
Concentration is abscissa, TGray value/CGray valueFor ordinate fit standard curve, and the T for passing through unknown sampleGray value/CGray valueIt obtains unknown
The data of Cloud Server are established rule by machine learning on computers by AI mode by sample concentration, compare position sample
Relevant information can obtain the concentration of unknown sample.
Embodiment 3
A kind of detection method of mycotoxin is the portable mycotoxin immunochromatography quantitative detection using embodiment 2
Device detects mycotoxin standard sample or sample to be tested；First pass through smart phone rear camera and micro-lens
The light path system of composition realizes the acquisition of image；APP inspection software is quantified by being built in the self-programming of smart phone again, to rear
The rgb signal for setting camera captured image is handled and is converted to corresponding numerical value；Finally by a series of inspection of concentration
It surveys, establishes standard curve, realize the quantitative detecting analysis to the middle mycotoxin of sample to be tested.
Wherein, quantitative APP inspection software includes two kinds of operational modes of artificial mode and AI mode；As the small Shi Caiyong of sample size
Artificial mode calculates T by the RGB information of one group of master sample image of analysisGray value/CGray value, and fit standard curve, it is fixed to realize
Amount detection；AI mode is used when sample size is big, and the data of Cloud Server are passed through by machine learning by AI mode on computers
Rule is established, the relevant information for comparing position sample obtains the concentration of unknown sample.
Above-mentioned detection method includes the following steps:
S1. the mycotoxin standard sample of equivalent various concentration is added into mycotoxin immunochromatographydetection detection card well
Product react 3~5min；
S2. operation mycotoxin immunochromatography quantitative detection APP Image Acquisition plate acquires image and is uploaded to cloud service
Device；
S3. operation mycotoxin immunochromatography quantitative detection APP image analysis plate artificial mode obtains sample RGB information
With examination criteria curve；If sample size is big, using AI mode；
S4. the appropriate mycotoxin sample to be measured of addition into mycotoxin immunochromatographydetection detection card well, reaction 3~
5min；
S5. operation mycotoxin immunochromatography quantitative detection APP Image Acquisition plate acquires image and is uploaded to cloud service
Device；
S6. by operation mycotoxin immunochromatography quantitative detection APP testing result plate color developing detection as a result, and uploading
As a result with sample geography information to network.
The sample of the portable mycotoxin immunochromatography quantitative testing device detection can be the paddy such as corn and soybean, wheat
Object and feed etc..Below by taking the detection of zearalenone as an example, it is specifically shown in embodiment 4.
Embodiment 4
The present embodiment is the detection method of zearalenone, is specifically comprised the following steps:
First sample to be tested is milled, weighs 1g powder, 50% methanol solution of 2mL is added, acutely vibrates 3min, Yu Changwen
Lower 5000rpm/min is centrifuged 5min, takes supernatant, and dilute 4 times with 0.02M PB solution (pH=7.4), is inhaled with rubber head dropper
2 drop supernatants are taken to be added in zearalenone immunochromatography quantitative test card well, layer is immunized in operation zearalenone
It is strong in the colour developing of zearalenone immunochromatographydetection detection card T, C line to analyse quantitative detection APP calculating various concentration zearalenone
Degree, and obtain zearalenone content and TGray value/CGray valueStandard curve, realize the quantitative detection of zearalenone, and
It is shared by wireless network that result can be will test.
Wherein, Portable corn zeranol immunochromatography quantitative testing device, including immunochromatography system, Image Acquisition
System, image reading analysis system.Immunochromatography system is zearalenone blue latex micro-ball immune chromatography quantitative detection
Card；Image capturing system is made up of photosensitive resin the camera bellows of 3D printing, handset bracket, micro-lens and smart phone；Figure
As acquisition analysis system is realized by zearalenone immunochromatography quantitative detection APP.
Zearalenone blue latex micro-ball immune chromatography quantitative test card by shell, bottom plate, sample pad, bonding pad,
Nitrocellulose filter (NC film), blotting paper composition.The zearalenone list that blue latex microballoon has marked is sprayed on bonding pad
Clonal antibody, drawing on NC film has zearalenone antigen ZEN-OVA and sheep anti-mouse igg, and antigen ZEN-OVA is T line, sheep anti mouse
IgG is C line.
(1) film is drawn
Antigen ZEN-OVA concentration is 0.1mg/mL, and sheep anti-mouse igg concentration is 0.2mg/mL, antigen ZEN-OVA and sheep anti mouse
IgG is diluted by PB solution (0.02M, pH=7.4), is drawn film instrument by metal spraying and is drawn antigen ZEN-OVA, sheep anti-mouse igg to NC
On film, dosage is 0.8 μ L/cm, and 37 DEG C of oven drying 2h after pulling are sealed spare.
(2) blue latex microballoon zearalenone antibody marks
Dissolution: being added 1mL MES solution (0.05M, pH=5.5) in 1.5mL centrifuge tube, and 0.5mg blue cream is then added
Glue microspheres solution (5mg/mL)；
Activation: 15 μ L 0.5mg/mL EDC solutions, 18 μ L 0.5mg/mL NHS solution are added in the above solution；
200rpm/min reacts 15min, then the 14000rpm/min at 4 DEG C, is centrifuged 15min；
Label: removal supernatant is added 1mL BB solution (0.05M, pH=8.0) and 1 μ L zearalenone Dan Ke is added
Grand antibody (5mg/mL) 200rpm/min reacts 30min；
Closing: 20 μ L 20%BSA solution are added in the above solution, 200rpm/min reacts 15min, then at 4 DEG C
14000rpm/min is centrifuged 15min；
Redissolve: removal supernatant is added 200 μ L and redissolves liquid, and 4 DEG C of preservations are stand-by.
Redissolve formula of liquid are as follows: 0.02M PB solution (pH=7.4) contains 0.5%Triton-100,5% trehalose, 0.5%
BSA, 0.5%PVP and 0.03%procline-300.
(3) ball is sprayed
The blue latex microspheres solution of upper zearalenone monoclonal antibody will be marked to draw film instrument by metal spraying with 3.5 μ
L/cm is sprayed on bonding pad, 37 DEG C of oven drying 30min, is sealed spare.
(4) detection card assembling
Stick dried NC film on PVC bottom plate, bonding pad is superimposed 1~2mm of NC film, sample pad be superimposed bonding pad 1~
2mm, blotting paper are superimposed 1~2mm of NC film.It is cut into 3.05mm with strip cutting machine after assembling, is fitted into getting stuck, desiccant is put into and waits for
With.
(5) sample pre-treatments
1g sample is weighed, 50% methanol solution of 2mL is added, acutely shakes 3min, 5000rpm/min is centrifuged under room temperature
5min collects supernatant, and dilutes 4 times for use with PB solution (0.02M, pH=7.4).
(6) sample is run
Assembled detection card well is added with the zearalenone sample that rubber head dropper draws 2 drop various concentrations
In, react 3~5min.
(7) Image Acquisition is analyzed
The jade of 2 drop various concentrations is added into zearalenone immunochromatography quantitative test card well with rubber head dropper
Zearlenone standard sample；After reacting 3~5min, zearalenone immunochromatography quantitative detection APP Image Acquisition is operated
Plate acquisition image is simultaneously uploaded to Cloud Server；Operate zearalenone immunochromatography quantitative detection APP image analysis plate
Artificial mode obtains sample RGB information and examination criteria curve, if sample is more, just uses AI mode；Rubber head dropper is to corn
2 drop zearalenone samples to be measured are added in zeranol immunochromatography quantitative test card well；After reacting 3~5min,
Operation zearalenone immunochromatography quantitative detection APP Image Acquisition plate acquisition image is simultaneously uploaded to Cloud Server；Pass through
Zearalenone immunochromatography quantitative detection APP testing result plate color developing detection is operated as a result, and uploading result and sample
Geography information is to network.
Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than protects to the present invention
The limitation of shield range can also be made on the basis of above description and thinking for those of ordinary skill in the art
Other various forms of variations or variation, there is no necessity and possibility to exhaust all the enbodiments.It is all of the invention
Made any modifications, equivalent replacements, and improvements etc., should be included in the protection of the claims in the present invention within spirit and principle
Within the scope of.
Claims (10)
1. a kind of mycotoxin immunochromatography quantitative test card (1), which is characterized in that including bottom plate (11), the bottom plate (11)
On be successively arranged sample pad (12), bonding pad (13), nitrocellulose filter (14) and blotting paper (15)；The blotting paper (15) and
Bonding pad (12) overlaps the both ends for being pressed in nitrocellulose filter (14) respectively, forms detection on the surface of nitrocellulose filter (14)
Area；The sample pad (12) is overlapping to be pressed on bonding pad (13)；It is micro- that colloidal gold, color latex are fixed on the bonding pad (13)
The mycotoxin antibody of ball or fluorescent microsphere label；Mycotoxin is fixed on nitrocellulose filter (14) in the detection zone
The nature controlling line that the detection line and sheep anti mouse or goat anti-rabbit igg secondary antibody that antigen is constituted are constituted.
2. mycotoxin immunochromatography quantitative test card according to claim 1, which is characterized in that the detection line is located at inspection
One end in area close to bonding pad (13) is surveyed, the nature controlling line is located at one end in detection zone close to blotting paper (15).
3. mycotoxin immunochromatography quantitative test card according to claim 1, which is characterized in that it further include shell (16),
The bottom plate (11), sample pad (12), bonding pad (13), nitrocellulose filter (14) and blotting paper (15) are placed in shell (16)
Interior, the position in shell (16) shell surface corresponding to nitrocellulose membrane (14) is equipped with observation window (17), in shell (16) shell surface
Position corresponding to sample pad (12) is equipped with well (18).
4. a kind of portable mycotoxin immunochromatography quantitative testing device, which is characterized in that any including claims 1 to 3
The item mycotoxin immunochromatography quantitative test card (1), sequentially connected camera bellows (2), micro-lens (3), handset bracket (4)
With smart phone (5), the mycotoxin immunochromatography quantitative test card (1) is embedded in camera bellows (2).
5. portable mycotoxin immunochromatography quantitative testing device according to claim 4, which is characterized in that the microspur
Camera lens (3) includes the first micro-lens (31) and the second micro-lens (32) being connected, the first micro-lens (31) it is another
End connects with the rear camera of smart phone (5), and the other end of the second micro-lens (32) connects with camera bellows (2).
6. a kind of detection method of mycotoxin, which is characterized in that be using the portable mycotoxin of claim 4 or 5
Immunochromatography quantitative testing device detects mycotoxin standard sample or sample to be tested；After first passing through smart phone (5)
Set the acquisition of the light path system realization image of camera and micro-lens (3) composition；Again by being built in smart phone (5)
Self-programming quantifies APP inspection software, is handled the rgb signal of rear camera captured image and is converted to corresponding number
Value；Finally by a series of detection of concentration, standard curve is established, realizes the quantitative detection point to mycotoxin in sample to be tested
Analysis.
7. detection method according to claim 6, which is characterized in that the quantitative APP inspection software include artificial mode and
Two kinds of operational modes of AI mode；Artificial mode is used when sample size is small, by the RGB information for analyzing one group of master sample image
Calculate TGray value/CGray value, and fit standard curve, realize quantitative detection；AI mode is used when sample size is big, it will by AI mode
The data of Cloud Server establish rule by machine learning on computers, and the relevant information for comparing position sample obtains unknown sample
Concentration.
8. detection method according to claim 7, which comprises the steps of:
S1. the mycotoxin standard of equivalent various concentration is added into mycotoxin immunochromatography quantitative test card (1) well
Sample reacts 3~5min；
S2. operation mycotoxin immunochromatography quantitative detection APP Image Acquisition plate acquires image and is uploaded to Cloud Server；
S3. operation mycotoxin immunochromatography quantitative detection APP image analysis plate artificial mode obtains sample RGB information and inspection
Mark directrix curve；If sample size is big, using AI mode；
S4. appropriate mycotoxin sample to be measured is added into mycotoxin immunochromatography quantitative test card (1) well (18), instead
Answer 3~5min；
S5. operation mycotoxin immunochromatography quantitative detection APP Image Acquisition plate acquires image and is uploaded to Cloud Server；
S6. by operation mycotoxin immunochromatography quantitative detection APP testing result plate color developing detection as a result, and uploading result
With sample geography information to network.
9. detection method according to claim 8, which is characterized in that the mycotoxin is zearalenone.
10. detection method according to claim 9, which is characterized in that the mycotoxin immunochromatographydetection detection card (1)
In, the zearalenone monoclonal antibody of blue latex microballoon label is fixed on the bonding pad (13)；The detection zone
The detection line and sheep anti-mouse igg two of zearalenone antigen ZEN-OVA composition are fixed on interior nitrocellulose filter (14)
The nature controlling line of anti-composition.
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Application publication date: 20190910 |