CN208654173U - T-2 toxin quick and quantitative determination system in a kind of food or feed - Google Patents

Abstract

The utility model discloses T-2 toxin quick and quantitative determination systems in a kind of food or feed, including chromatography detection card, chromatogram scanner, client and cloud platform；The chromatogram scanner is scanned for carrying chromatography detection card, and to chromatography detection card；The chromatogram scanner is wirelessly connected client；Client connects cloud platform by internet.Coating BSA-T2 haptens is detection line on the nitrocellulose filter of the chromatography detection card, and coating goat-anti chicken antibody is nature controlling line.The utility model high sensitivity, batch internal difference and difference between batch of detection are small, have preferable repeatability, performance stabilization is, it can be achieved that T-2 toxin field quick detection, practical value with higher.

Description

T-2 toxin quick and quantitative determination system in a kind of food or feed

Technical field

The utility model relates to a kind of detection systems, more particularly to T-2 toxin fast quantification in a kind of food or feed Detection system.

Background technique

T2 toxin is a kind of fusarium fungus (mainly metabolism of fusarium tricinctum (Fusarium trieinetum) Product is to belong to a kind of strongest toxin of Trichothecenes mycin toxicity, is widely present in cereal crops and its semi-finished product, people It after eating the cereal of pollution by mistake, can cause vomiting, the acute poisonings symptom such as diarrhea, fever, hemopoietic system be damaged when serious, at people and animals It is dead.Studies have shown that T-2 toxin shows a variety of toxicity such as genotoxicity, immunotoxicity, hematotoxicity to humans and animals and can Cause apoptosis of many kinds.FAO (Food and Agriculture Organization of the United Nation)s (FAO) in 1973 and the World Health Organization (WHO) hold joint in Geneva In meeting, using T-2 toxin with aflatoxin equally as the most dangerous food pollution source of naturally occurring.

T-2 toxin is the trichothecene that nature is found earliest, shows as stronger toxicity.Research at present Show that the mankind have 3 kinds of endemic diseases: the cause of disease of alimentary toxic aleukia, Kaschin-Beck disease and Keshan disease be considered with T-2 toxin is related.Food poisoning aleucemia takes place mostly in the western some areas in Siberia, and China then mainly occurs Kaschin-Beck disease and Keshan disease, wherein Kaschin-Beck disease is mainly distributed on the area from northeast Heilongjiang Province to Tibet Autonomous Region in China On.In addition, T-2 toxin is also related with the high incidence of China certain areas cancer of the esophagus.T-2 toxin toxicological profile mainly shows To cause vomiting, heartbeat is slow, diarrhea, bleeding, oedema, skin histology necrosis, gastrointestinal tract mucosa bleeding, hematopoietic tissue destroy and Immunosupress, peripheral white blood cells and decrease of platelet, meningorrhagia, nerve problems, food refusal or anorexia, cardiovascular system Destroy etc..

Vomitoxin detection discovery generates extensively in the world, all detects from the grain sample acquired all over the world To this kind of toxin.Wherein the recall rate in China's cereal is 80%.Corn, wheat, barley, oat, rye have by T-2 toxin Polluting the most serious crop varieties, the T-2 toxin remained in animal product can also pass to consumer by food chain, and one The government of a little countries has taken strong Supervision Measures, to eliminate consumer to the worry of pork product safety.For example, European The highest that T-2 in all food was set up in 1997 allows content to be 5ppb.This standard is tighter set to by Germany 3ppb.Currently on the market using the enzyme Labeling Kit of detection T-2 toxin, but ELISA kit needs microplate reader, incubation reaction item Part, board-washing condition, operation element environment and professional technician；And quickly colloidal gold is although easy to detect, is not required to professional technique Personnel and working environment, but the sensitivity of its detection is low, and can only be qualitative, and detection range is narrow.And the mesh of the utility model patent Be exactly the respective disadvantage for overcoming the above two classes kit, by shirtsleeve operation, scene quickly, it is accurate, quantitative with sensitivity (sxemiquantitative) analysis, realizes the on-site test in food and feed.

Utility model content

The purpose of this utility model is just to provide T-2 toxin quick and quantitative determination system in a kind of food or feed, can be complete Complete solution is determined in place of above-mentioned the deficiencies in the prior art.

The purpose of this utility model is realized by following technical proposals:

T-2 toxin quick and quantitative determination system in a kind of food or feed, including chromatography detection card, chromatogram scanner, visitor Family end and cloud platform；The chromatogram scanner is scanned for carrying chromatography detection card, and to chromatography detection card；The chromatography Scanner is wirelessly connected client；Client connects cloud platform by internet, and the cloud platform is for storing tested T-2 toxin Standard curve and detection data；The chromatography detection card includes the detection card ontology of assembly inside the shell, detection Ka Bentibao Bottom plate is included, sample pad, label pad, nitrocellulose filter and blotting paper are set gradually on bottom plate, the label pad is by carrier base Layer and marker composition, marker are that spraying lanthanide fluoro detects microballoon in carrier substrate and lanthanide fluoro Quality Control microballoon is formed One tunic, the detection microballoon and Quality Control microballoon are surface with carboxyl polystyrene microsphere, and detection microballoon and Quality Control are micro- The partial size of ball is 50~500nm, and it is detection line that BSA-T2 haptens is coated on the nitrocellulose filter, and coating goat-anti chicken is anti- Body is nature controlling line, and counter sample pad opens up well on the shell, and the detection line and nature controlling line on corresponding nitrocellulose filter are opened If form hole；The client is built-in with receiving module, data processing module, memory module and communication module, the receiving module, Memory module and communication module are connect with data processing module；Receiving module is for receiving the number that chromatogram scanner is transmitted to According to, and pass to data processing module；Data processing module carries out data analysis and passes the result to memory module and communication Module；Communication module connects cloud platform by internet.

Preferably, the bottom plate be PVC bottom plate, on PVC bottom plate from left to right successively paste sample pad, label pad, Nitrocellulose filter and blotting paper, and the right end of sample pad rides on the left end of label pad, the right end of label pad rides over nitric acid fibre On the left end for tieing up plain film, blotting paper left end is ridden on the right end of nitrocellulose filter.

Preferably, the detection microballoon is the microballoon for being marked with the monoclonal antibody of T-2 toxin, Quality Control microballoon is to be marked with chicken The Quality Control microballoon of lgY or rabbit lgG；The goat-anti chicken antibody that nature controlling line uses on the nitrocellulose filter is the lgG of goat-anti chicken lgY Or goat-anti rabbit lgG.

Preferably, the lanthanide fluoro of the lanthanide fluoro microballoon of the label on biological raw material refer to lanthanide series Eu, The fluorescence that Sm or Tb and beta-diketone ligand generate.Its luminous efficiency is high, absorption spectrum is wide, emission spectrum is very narrow, this Tekes position Width is moved, keeps the sensitivity of its spectrofluorimetry very high.

Preferably, the client is with Internet enabled mobile phone or tablet computer.

Explanation of nouns:

The lgG of goat-anti chicken lgY: the immunoglobulin G of goat-anti chicken lgY；

Goat-anti rabbit lgG: the immunoglobulin G of goat-anti rabbit lgG；

Chicken lgY: chicken yolk antibody；

Rabbit lgG: rabbit immunoglobulin G；

BSA-T2 haptens: T-2 toxin couples the polymer composite to be formed by chemical combination key and BSA.

Compared with prior art, the utility model has the beneficial effects that:

1. it is easy to operate, it is able to achieve live quick, accurate, quantitative (sxemiquantitative) analysis with sensitivity, realizes food and feeding The on-site test of material；

2. high sensitivity, batch internal difference and difference between batch of detection are small, there is preferable repeatability, and performance is stablized, yellow bent with AFB1( Mould toxin B1), ZEN(zeranol), DON(vomitoxin), FB1(fumonisin B1), OTA(ochratoxin A) etc. moulds Toxin does not have cross reaction.

3. T-2 toxin field quick detection can be realized, statistics, management, practical valence with higher are realized by cloud platform Value.

Detailed description of the invention

Fig. 1 is the functional block diagram of the utility model embodiment one；

Fig. 2 is the structural schematic diagram of chromatography detection card in the utility model embodiment one；

Fig. 3 is the structural schematic diagram that card ontology is detected in Fig. 1；

Fig. 4 is the top view of Fig. 3；

Fig. 5 is T-2 Mycotoxin identification standard curve；

Fig. 6 is the structural schematic diagram of chromatography detection card in the utility model embodiment two.

Specific embodiment

The utility model is further described with attached drawing combined with specific embodiments below.

Embodiment one

As shown in Figures 1 to 5, T-2 toxin quick and quantitative determination system in a kind of food or feed, including chromatography detection Card, chromatogram scanner, client and cloud platform.

The chromatogram scanner is scanned for carrying chromatography detection card, and to chromatography detection card；The computed tomography scanning Instrument is wirelessly connected client；Client connects cloud platform by internet, which is used to store the standard of tested T-2 toxin Curve and detection data.

The chromatography detection card includes the detection card ontology being assemblied in shell 10, and the detection card ontology includes the bottom PVC Plate 1 successively pastes sample pad 2, label pad 3, nitrocellulose filter 4 and blotting paper 5, and sample from left to right on PVC bottom plate 1 The right end of pad 2 rides on the left end of label pad 3, and the right end of label pad 3 rides on the left end of nitrocellulose filter 4, and blotting paper 5 is left End rides on the right end of nitrocellulose filter 4.It is detection line 6, coating that BSA-T2 haptens is coated on the nitrocellulose filter 4 Goat-anti chicken antibody is nature controlling line 7.Counter sample pad 2 opens up well 11, the inspection on corresponding nitrocellulose filter 4 on shell 10 Survey line 6 and nature controlling line 7 open up form hole 12.

The goat-anti chicken antibody that nature controlling line 4 uses on the nitrocellulose filter 4 is the lgG of goat-anti chicken lgY or goat-anti rabbit lgG。

The label pad 3 is made of carrier substrate and marker, and marker is spraying lanthanide fluoro detection in carrier substrate The tunic that microballoon and lanthanide fluoro Quality Control microballoon are formed, and detecting microballoon is the microballoon for being marked with T-2 toxin antibody, Quality Control is micro- Ball is the Quality Control microballoon for being marked with chicken lgY or rabbit lgG.The detection microballoon and Quality Control microballoon are surface with carboxyl polyphenyl second Alkene microballoon.Also, the partial size for detecting microballoon and Quality Control microballoon is 50~500nm.

The lanthanide fluoro microballoon sprayed in label pad 3, it is glimmering that lanthanide fluoro refers to that lanthanide series is generated with beta-diketone ligand Light marks the lanthanide fluoro substance of the lanthanide fluoro microballoon on biological raw material to refer to lanthanide series Eu, Sm and Tb and beta-diketon The fluorescence that aglucon generates.Using unique Fluorescence Characteristic of detector test lanthanide series and complex.

When carrying out chromatography detection, the marker and nitrocellulose of T-2 toxin, T-2 toxin antibody in food and feed Coated BSA-T2 haptens detects the T-2 toxin in food and feed by striving method unexpectedly on film.

The client is built-in with receiving module, data processing module, memory module and communication module, the reception mould Block, memory module and communication module are connect with data processing module；Receiving module is transmitted to for receiving chromatogram scanner Data, and pass to data processing module；Data processing module carry out data analysis and pass the result to memory module and Communication module；Communication module connects cloud platform by internet.The client is with Internet enabled mobile phone or tablet computer.

Working principle:

It after sample to be tested is carried out pre-treatment, instills in the well of chromatography detection card, chromatography examined chromatography after 15 minutes Survey card is put into chromatogram scanner and is scanned, and the excitation light source in chromatogram scanner issues exciting light, blocks to chromatography detection and carries out Irradiation, so that the fluorescent microsphere on chromatography detection card is excited and generates fluorescence, chromatogram scanner is acquired point by point on chromatography detection card Fluorescence signal intensity value, and then complete the detection of entire chromatography detection card.The chromatogram scanner will test data and pass through indigo plant Tooth passes to the receiving module of client, fluorescence signal intensity detection data of the data processing module according to chromatography detection card, meter Calculate the fluorescence signal intensity value that chromatography detection blocks upper nature controlling line and detection line, at the same data processing module by communication module from Cloud platform obtains the standard curve of the T-2 toxin of sample, and is stored in memory module, and data processing module is according to standard song Line (contrast relationship of the fluorescence signal intensity value and standard curve of nature controlling line and detection line), is calculated T-2 in sample The content of toxin, and shown, testing result is stored in memory module and cloud platform.

Prepare T-2 Mycotoxin identification standard curve:

Configure 6 parts of the calibration solution (standard items of toxin containing T-2) containing T-2 toxin, concentration is respectively 0,100,250,100, 2500,5000(T-2 toxin standard items).The calibration solution of above-mentioned various concentration is separately added into adding for the chromatography detection card assembled In sample hole, chromatography is detected by chromatogram scanner after 15 minutes, the testing result linear regression that 6 times obtain is marked Directrix curve, referring to fig. 4.

The application method of the chromatography detection card: after comminuted sample, powder is sifted out using not less than 20 meshes.Weigh 1g Sample is crushed in 10mL centrifuge tube, adds 5mL sample extracting solution (or equal proportion increases sample and extracting solution), vibrates 5 minutes.Room Temperature centrifugation 5 minutes, or stand and supernatant is presented.Clean 1.5mL centrifuge tube is taken, the dilution of 100 μ L and 400uL sample of supernatant is added Liquid vibrates 1 minute.The sample 80ul diluted is added in well 11, under the action of blotting paper 5, sample is from sample pad 2 It is mobile to 5 direction of blotting paper.15min is chromatographed in the case where avoiding strong illumination, and then chromatography is detected with chromatogram scanner Card is detected, standard curve referring to fig. 4.

Above-mentioned chromatography detection card the preparation method is as follows:

Step 1 is pasted onto nitrocellulose filter on PVC bottom plate, using film metal spraying special purpose machinery in nitrocellulose The lgG formation nature controlling line for being diluted to the goat-anti chicken lgY of 0.3mg/ml and the BSA-T2 for being diluted to 0.2mg/ml are sprayed on film Haptens formed detection line, discharge rate be 1 μ l/cm, then 37 DEG C at a temperature of bake 8 hours；

Step 2, the lanthanide fluoro for preparing the lanthanide fluoro microballoon for being marked with chicken lgY and being marked with T-2 toxin antibody are micro- The lanthanide fluoro microballoon of 1mL is added to the MES(2-(N- morpholine of 50mg by ball) ethanesulfonic acid) buffer (0.1M, pH7.0) In, add 10mg carbodiimide (EDC) and 10mg n-hydroxysuccinimide sulfonate sodium stirring and dissolving, room temperature reaction 30 Centrifugally operated is carried out after minute, centrifugal sediment is redissolved with 50mM borate buffer (pH8.2), the chicken that 2mg dialysed is added LgY is stirred to react 24 hours at room temperature, after being then centrifuged for, closing, then (Conservation environment temperature is saved in dilution It is 2~8 DEG C) to get the lanthanide fluoro microballoon for being marked with chicken lgY；Using the lanthanum of above-mentioned same method label T-2 toxin antibody It is fluorescent microsphere；

Step 3, being marked with chicken lgY and be marked with the lanthanide fluoro microballoon of T-2 antibody and be diluted to 0.1 μ of concentration respectively G/ml and 3 μ g/ml is sprayed in carrier substrate using a film metal spraying machine and constitutes label pad, and discharge rate is 2.5 μ l/cm, Then 37 DEG C at a temperature of bake 8 hours；

Step 4 is successively sticked to sample pad, label pad, blotting paper on PVC bottom plate, is assembled into kilocalorie, then use cutter The test strips of 5mm wide are cut into, are assemblied in detection card shell to get this T-2 Mycotoxin identification card is arrived.

Embodiment two

Referring to Fig. 6, the present embodiment and embodiment one the difference is that, 10 surface pair of shell of the chromatography detection card It answers the position of blotting paper 5 to offer multiple through-holes 13, is conducive to solvent and volatilizes shell 10, it is ensured that testing result is accurate, solves The technical issues of detection card shell of the prior art uses enclosed construction, and solvent can not volatilize away.

In order to guarantee volatilization effect, equidistantly uniformly opened in the region that 10 surface of shell matches with 5 size of blotting paper If through-hole 13.

Embodiment three

The present embodiment and embodiment one or two the difference is that, the detection card ontology tilts and is fixed on shell 10 It is interior, and sample pad 2 is in low side, blotting paper 5 is in high-end.This structure makes the sample to be tested flow velocity instilled from well 11 Slow down, slowly up chromatograph, guarantee the enough chromatography time, can effectively prevent liquid sample flow it is too fast and influence detection knot Fruit avoids the too fast inflow nitrocellulose filter 4 of sample that testing result is caused to fail.Because holding if sample to be tested flow velocity is too fast It is insufficient to easily lead to chromatography, and then influences testing result.It is specific according to the difference of sample to be tested to detect the inclined angle of card ontology It is designed.

Example IV

The present embodiment and embodiment one the difference is that, be equipped with holding tank in the shell 10 of the chromatography detection card, And 2 holding tanks are symmetrically arranged in the two sides of detection card ontology, sheet desiccant is installed in holding tank.It is this design so that Desiccant keeps together with detection card ontology, so that detection card ontology humidity is effectively controlled, to improve detection accuracy.

As a preference, sample reserve tank with cover can also be set on 10 surface of shell of chromatography detection card, it can be with Seal sample up for safekeeping.The hd top face of the sample reserve tank is flushed with 10 surface of shell, is covered equipped with pit, is applied convenient for users to finger Power is turned-out to uncap.

The above is only the preferred embodiment of the utility model only, is not intended to limit the utility model, all at this Made any modifications, equivalent replacements, and improvements etc., should be included in the utility model within the spirit and principle of utility model Protection scope within.

Claims (2)

1. T-2 toxin quick and quantitative determination system in a kind of food or feed, it is characterised in that: including chromatography detection card, chromatography Scanner, client and cloud platform；

The chromatogram scanner is scanned for carrying chromatography detection card, and to chromatography detection card；

The chromatogram scanner is wirelessly connected client；

Client connects cloud platform by internet, which is used to store the standard curve and testing number of tested T-2 toxin According to；

The chromatography detection card includes the detection card ontology of assembly inside the shell, which includes bottom plate, on bottom plate Sample pad, label pad, nitrocellulose filter and blotting paper are set gradually, the label pad is made of carrier substrate and marker, Marker is the tunic that spraying lanthanide fluoro detects that microballoon and lanthanide fluoro Quality Control microballoon are formed in carrier substrate, the detection Microballoon and Quality Control microballoon are surface with carboxyl polystyrene microsphere, and the partial size for detecting microballoon and Quality Control microballoon be 50~ 500nm, coating BSA-T2 haptens is detection line on the nitrocellulose filter, and coating goat-anti chicken antibody is nature controlling line, outside Counter sample pad opens up well on shell, and the detection line and nature controlling line on corresponding nitrocellulose filter open up form hole；The layer The position that the case surface of analysis detection card corresponds to blotting paper offers multiple through-holes, and the case surface setting of chromatography detection card is with cover Sample reserve tank, detection card ontology inclination is fixed inside the shell, and sample pad be in low side, and blotting paper is in high-end；

The client is built-in with receiving module, data processing module, memory module and communication module, the receiving module, storage Module and communication module are connect with data processing module；Receiving module is used to receive the data that chromatogram scanner is transmitted to, And pass to data processing module；Data processing module carries out data analysis and passes the result to memory module and communication mould Block；Communication module connects cloud platform by internet；

The bottom plate be PVC bottom plate, on PVC bottom plate from left to right successively paste sample pad, label pad, nitrocellulose filter and Blotting paper, and the right end of sample pad rides on the left end of label pad, the right end of label pad rides on the left end of nitrocellulose filter, Blotting paper left end rides on the right end of nitrocellulose filter；

The detection microballoon is the microballoon for being marked with the monoclonal antibody of T-2 toxin, and Quality Control microballoon is the matter for being marked with chicken lgY or rabbit lgG Control microballoon；The goat-anti chicken antibody that nature controlling line uses on the nitrocellulose filter is the lgG of goat-anti chicken lgY or goat-anti rabbit lgG；

The client is with Internet enabled mobile phone or tablet computer.

2. T-2 toxin quick and quantitative determination system in food according to claim 1 or feed, it is characterised in that: described The lanthanide fluoro of the lanthanide fluoro microballoon on biological raw material is marked to refer to that lanthanide series Eu, Sm or Tb and beta-diketone ligand are produced Raw fluorescence.