CN206594181U - A kind of vomitoxin direct competive ELISA kit - Google Patents
A kind of vomitoxin direct competive ELISA kit Download PDFInfo
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- CN206594181U CN206594181U CN201720173786.6U CN201720173786U CN206594181U CN 206594181 U CN206594181 U CN 206594181U CN 201720173786 U CN201720173786 U CN 201720173786U CN 206594181 U CN206594181 U CN 206594181U
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- reagent bottle
- vomitoxin
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Abstract
The utility model discloses a kind of vomitoxin direct competive ELISA kit, belong to immunochemistry detection technique field.The technical solution of the utility model main points are：A kind of vomitoxin direct competive ELISA kit, including box body, lid and it is arranged at the fixed seat of box body inside bottom, fixed seat is provided with the recessed bottle position of reagent bottle, it is respectively used to place in concentrated cleaning solution reagent bottle, sample diluting liquid reagent bottle, ELIAS secondary antibody working solution reagent bottle, substrate developer A liquid reagent bottle, substrate developer B liquid reagent bottle, terminate liquid reagent bottle and standard solution reagent bottle, box body and is also placed with ELISA Plate, micro pipette, centrifuge tube, colorimetric card, specification and cover plate film.The utility model carries out the qualitative and quantitative analysis of vomitoxin using direct competive ELISA pattern, has the advantages that simple to operate, sensitivity is high, with low cost, detecting step is few and detection time is short, is adapted to the field quick detection of batch samples.
Description
Technical field
The utility model belongs to immunochemistry detection technique field, and in particular to a kind of vomitoxin direct competive ELISA
Kit.
Background technology
Vomitoxin（vomitoxin）Also known as deoxynivalenol（Deoxynivalenol, DON）, with very
Strong cytotoxicity, can cause animal bowel dysfunction, mucous membrane of mouth and dermal lesions, immunity degradation.People and livestock are eaten by mistake
Produced extensive poisonous effect after the Cereals of endotoxin contamination, can cause vomiting, food refusal, the symptom such as weight loss, it is immune and make
Blood system suffers damage, and can damage hemopoietic system also with very strong dermal toxicity and cytotoxicity, when serious causes death.
It is distributed in DON in the cereal seed such as wheat, barley, corn more, is the mycotoxin that most serious is polluted after aflatoxin.
Because DON harm is serious, the most attention of various countries is caused, the strict of DON residual quantities in cereal and feed has been formulated in succession
Limit standard.DON limit standard is in China's cereal, swine feed, calf mixed feed, lactation period animal mixed feed
DON limit standard is 5.0mg/kg in 1.0mg/kg, ox mixed feed, poultry mixed feed.
The conventional method of DON residue detections has microbiology method, fluorescence method, thin-layered chromatography (TLC), high-efficient liquid phase color
Spectrometry（HPLC）, and Liquid Chromatography/Mass Spectrometry（LC-MS）, gas chromatography mass spectrometry detection method（GC-MS）And Liquid Chromatography-Tandem Mass Spectrometry
（LC-MS/MS）Etc. method.These method sensitivity are high, qualitative accurate, it may be determined that the molecular weight of compound, molecular formula, even
Functional group, is adapted to quantitatively detect the compound and its metabolin.But these method costs are high, it is necessary to which technical professional, holds high
Expensive laboratory apparatus and prolonged preliminary preparation.And immunological detection method is set up in molecular recognition of the antibody to antigen
On, its major advantage is that affinity is high, quick, simple, economical and practical, can realize small size, the big flux to biofluid
Detection, is increasingly becoming one of method of countries in the world noxious residual chemicals quick detection.
Therefore, quick, sensitive, efficient vomitoxin direct competive ELISA kit is developed for ensureing animality food
Tool is of great significance product safely.
The content of the invention
The technical problem that the utility model is solved there is provided a kind of vomitoxin direct competive ELISA examination of small volume and less weight
Agent box, easy to operate using the kit, sensitivity is high, and cost is low, and stability is good, and detecting step is few, and detection time is short, can enter
The measure of vomitoxin content, reduces the time required for sample detection in the multiple samples of row one-time continuous.
The utility model is to solve above-mentioned technical problem to adopt the following technical scheme that, a kind of vomitoxin direct competitive
ELISA kit, including box body, be connected to the lid of box body side, be arranged at the fixed seat of box body inside bottom and be arranged at
ELISA Plate, micro pipette, centrifuge tube, colorimetric card, specification and the cover plate film of tray interior, it is characterised in that：Described ELISA Plate
It is made up, using 96 hole agent plates as solid phase carrier, is put in aluminide-coating bag of polystyrene, and ELISA Plate is by outer frame support
With the dismantled and assembled enzyme mark reaction capillary strip composition being placed on it, each dismantled and assembled enzyme mark reaction capillary strip has 12 reacting holes, its
Upper pre-coated emesis toxin monoclone antibody；Described micro pipette has four kinds of different capabilities, the wherein micro pipette of 1.0mL capacity
30 are used for the sampling of sample substrate, and the micro pipette 10 of 100 μ L capacity is used to add terminate liquid, the micro pipette 20 of 50 μ L capacity
Root is used to add ELIAS secondary antibody working solution, and the micro pipette 40 of 30 μ L capacity is used to add substrate developer A liquid and substrate colour developing
Agent B liquid；Described centrifuge tube has three kinds of different capabilities, respectively 10mL, 5mL and 1.5mL, for locating before sample collection and matrix
Reason；Described colorimetric card has 5, the corresponding absorbance of display different colours reagent wells, for water sample, dairy produce, tealeaves, feeding
The reckoning of vomitoxin residual content in material and food；Described specification is used to introduce the principle of kit, standard curve, fitted
With scope, cross reacting rate, required instrument and reagent, application method and points for attention；Described cover plate film 5, for sample
ELISA Plate is covered during detection to prevent liquid evaporation from influenceing testing result；Described fixed seat is provided with 12 recessed bottles of reagent bottle
Position and the recessed bottles position of 2 standby reagent bottles, wherein the recessed bottle position of 12 reagent bottles be respectively used to place concentrated cleaning solution reagent bottle,
Sample diluting liquid reagent bottle, ELIAS secondary antibody working solution reagent bottle, substrate developer A liquid reagent bottle, substrate developer B liquid reagents
Bottle, terminate liquid reagent bottle, 200ng/mL vomitoxin standard solution reagent bottle, 100ng/mL vomitoxin standard items it is molten
Liquid reagent bottle, 50ng/mL vomitoxin standard solution reagent bottle, 25ng/mL vomitoxin standard solution reagent bottle,
12.5ng/mL vomitoxin standard solution reagent bottle and 6.25ng/mL vomitoxin standard solution reagent bottle, 2
The standby recessed bottle position of reagent bottle is respectively used to place high standard product solution reagent bottle, dilute acid soln reagent bottle, dilute alkaline soln
Reagent bottle or Tween-20 solution reagent bottle.
The utility model different size, the plastic bottle of different colours contain liquid and are fixed on consolidating for box body inside bottom
In reservation, together it is encapsulated in ELISA Plate, micro pipette, centrifuge tube, colorimetric card, specification and cover plate film in kit, is easy to take
Band and transport.Reagent in each kit carries out 96 parts of measure enough, you can carry out the detection of the multiple samples of one-time continuous,
Also ELISA Plate capillary strip can be taken apart and is used for multiple times.Now, remaining capillary strip is put back in aluminium foil bag and sealed with valve bag, this
Sample can ensure the Stability and veracity of kit testing result.
Kit of the present utility model is designed according to antigen-antibody competitive immunization chromatographic theory, on ELISA Plate capillary strip
The vomitoxin and ELIAS secondary antibody direct competitive polluted in pre-coated emesis toxin monoclone antibody, sample is coated on ELISA Plate
Monoclonal antibody on capillary strip, vomitoxin pollutant load is higher, and ELIAS secondary antibody and vomitoxin monoclonal antibody are combined
Quantity is fewer, and when adding substrate chromogenic reagent solution, sample absorbance value and vomitoxin content are negatively correlated, with standard curve
Compare multiplied by with its corresponding extension rate, you can draw the residual content of vomitoxin in sample.
The utility model has the advantages that compared with prior art：
（1）Operating procedure of the present utility model includes addition sample and ELIAS secondary antibody, addition substrate nitrite ion and sulfuric acid are whole
Only liquid, detecting step is few, detection time is short, than the time that traditional indirect competitive ELISA kit saves 30min, with tradition reason
Change detection method（1d）Compare, advantage becomes apparent；
（2）The utility model is the analytical technology based on the specificity of antigen-antibody molecule, invertibity reaction, is had
The selectivity and sensitiveness of height, and with the advantage such as easy, quick, screening amount is big, represent vomitoxin residue detection
Developing direction；
（3）The utility model is economical and practical, ageing strong, can carry out Site Detection, compared with Physico-chemical tests method, detection
1/20 of cost less than physico-chemical analysis method；
（4）The utility model is mainly used in vomitoxin pollution residue detection in water sample, tealeaves, feed and food, both may be used
For batch samples screening, the quick detection of small lot sample can be carried out again, for improving food security, ensureing people's body and mind
Health, holding are environment-friendly with realizing that sustainable development has important practical significance；
（5）The utility model is easy to a wide range of popularization and application, and is adapted to different classes of librarian use, and such as laboratory is examined
Survey, customs quarantine control, health supervision, breeding scale and individual production etc., with wide market prospects and significant economic benefit
And social benefit.
Brief description of the drawings
Fig. 1 is structural representation of the present utility model；
Fig. 2 is the structural representation of fixed seat in the utility model.
In figure：1st, box body, 2, lid, 3, fixed seat, 4, concentrated cleaning solution bottle position, 5, sample diluting liquid bottle position, 6, enzyme mark
Secondary antibody working solution bottle position, 7, substrate developer A liquid bottle position, 8, substrate developer B liquid bottle position, 9, terminate liquid bottle position, 10, standard items
Solution bottle position, 11, standby reagent bottle recessed bottle position.
Embodiment
Particular content of the present utility model is described in detail with reference to accompanying drawing.A kind of vomitoxin direct competive ELISA kit,
Including box body 1, it is connected to the lid 2 of the side of box body 1 and the fixed seat 3 of the inside bottom of box body 1 is set, described lid 2 is covered
The top of box body 1 and front side portion, border width protrude 0.3-0.5cm than box body 1, for switching kit.
Described fixed seat 3 is provided with the recessed bottle position of 12 reagent bottles and 2 standby recessed bottle positions 11 of reagent bottle, and concentration is washed
Wash liquid bottle position 4 and be placed with the 100mL concentrated cleaning solutions that bottom shape encapsulates for rectangle, translucent plastic reagent bottle（10×）, sample
It is rectangle, the 100mL samples dilution of the translucent plastic reagent bottle encapsulation of blue cap that product dilution bottle position 5, which is placed with bottom shape,
Liquid, ELIAS secondary antibody working solution bottle position 6 is placed with the 7mL sheep anti mouse ELIAS secondary antibodies of red plastic reagent bottle encapsulation（GaMIgG-
HRP）, substrate developer A liquid bottle position 7 is placed with the 4mL substrate developer A liquid of black plastic reagent bottle encapsulation, substrate developer B
Liquid bottle position 8 is placed with the 4mL substrate developer B liquid of green plastic reagent bottle encapsulation, and terminate liquid bottle position 9 is placed with yellow plastic examination
The 7mL 2M sulfuric acid solutions of agent bottle encapsulation, standard solution bottle position 10 is placed with the 1mL vomitings of opalescent plastic reagent bottle encapsulation
Totally 6 bottles of toxin standard solution, concentration be respectively 200ng/mL, 100ng/mL, 50ng/mL, 25ng/mL, 12.5ng/mL and
6.25ng/mL, the standby recessed bottle position 11 of reagent bottle is used for high standard product solution reagent bottle, the dilute acid soln reagent placed
Bottle, dilute alkaline soln reagent bottle, Tween-20 solution reagent bottle or particular sample treatment fluid reagent bottle.
ELISA Plate, micro pipette, centrifuge tube, colorimetric card, specification and cover plate are also placed with the box body 1 of the kit
Film, described ELISA Plate is made up of polystyrene, using 96 hole agent plates as solid phase carrier, is put in aluminide-coating bag, and enzyme
Target is made up of outer frame support and the dismantled and assembled enzyme mark being placed on it reaction capillary strip, each dismantled and assembled enzyme mark reaction capillary strip
There are 12 reacting holes, thereon pre-coated emesis toxin monoclone antibody.Described micro pipette has four kinds of different capabilities, wherein
The micro pipette 30 of 1.0mL capacity is used for the sampling of sample substrate, and the micro pipette 10 of 100 μ L capacity is used to add terminate liquid,
The micro pipette 20 of 50 μ L capacity is used to add ELIAS secondary antibody working solution, and the micro pipette 40 of 30 μ L capacity shows for adding substrate
Toner A liquid and B liquid.Described centrifuge tube has three kinds of different capabilities, respectively 10mL, 5mL and 1.5mL, is mainly used in sample and adopts
Collection and matrix pre-treatment.Described colorimetric card has 5, the corresponding absorbance of display different colours reagent wells, for water sample, milk
The general reckoning of vomitoxin residual content in product, tealeaves, feed and food.1 part of described specification, is discussed in detail reagent
The principle of box, standard curve, the scope of application, cross reacting rate, required instrument and reagent, application method and points for attention.It is described
Cover plate film 5, cover ELISA Plate during for sample detection, prevent liquid evaporation, influence testing result.
Vomitoxin residual content is determined in sample substrate：
1）Sample pre-treatments：Peanut, corn, wheat, rice and its product or feed 25g are weighed, 10g is added after crushing and is gathered
Ethylene glycol 8000, uses deionized water constant volume, is transferred in homogeneous cup, homogenizer high speed homogenization 2min, and then 4000r/min is centrifuged
5min is filtered with quantitative filter paper, takes supernatant to carry out sample detection.
2）The operating procedure of kit：1st, required reagent is taken out from cold storage environment, is placed in room temperature（20-25℃）Balance
30min, notices that every kind of liquid reagent must shake up before；2nd, the capillary strip of requirement is taken out, no capillary strip is put
Enter in former Fresco Bag and resealed together with the drier of offer, be stored in 2-8 DEG C；3rd, number：By sample correspondence micropore
Sequentially number, each sample does that 3 holes are parallel, and records the position where sample well；4th, product are loaded：The μ L of sample treatment liquid 50 are added
Enter in ELISA Plate capillary strip, while adding the μ L of ELIAS secondary antibody working solution 50, mix and stand board-washing after 15min, notice that detection sets three
Individual repetition, and set NC and BC to compare；5th, develop the color：Plus substrate developer A liquid and each 30 μ L of substrate developer B liquid, stand 5min
Board-washing afterwards；6th, terminating reaction：Add H2SO4The μ L of terminate liquid 100, stand ELIASA after 5min and read absorbance.
3）Vomitoxin residual content in determination sample matrix：
1st, qualitative determination：After substrate colour developing 5min, the blue depth of sample well is contrasted with colorimetric card, obtained in sample substrate
Vomitoxin residual content probable ranges；
2nd, quantitative determine：Add terminate liquid to stand after 5min, the absorbance that ELIASA is determined is substituted into specification
Standard curve, can vomitoxin residual content in Accurate Determining sample substrate.
4）Points for attention：
1st, room temperature is not returned to room temperature less than 20 DEG C or reagent and sample（20-25℃）Absorbance can be caused relatively low；
2nd, such as occur the situation of plate hole drying during board-washing, standard curve can be caused non-linear, repeatability is bad
Phenomenon；
3rd, without using the kit for spending validity date, do not exchange using the reagent in different lot number kits；
4th, during standard items, ELIAS secondary antibody and developer are to photaesthesia, therefore constant-temperature incubation, it is to avoid light irradiates, use
Cover plate film seals microwell plate；
5th, reaction terminating liquid is 2M sulfuric acid, it is to avoid contact skin；
6th, developing time by specification requires to carry out, and otherwise standard curve can be caused to be misfitted with actually detected data.
General principle of the present utility model, principal character and advantage has been shown and described above, it is new not departing from this practicality
On the premise of type spirit and scope, the utility model also has various changes and modifications, and these changes and improvements both fall within requirement and protected
The scope of the present utility model of shield.
Claims (1)
1. a kind of vomitoxin direct competive ELISA kit, including box body, it is connected to the lid of box body side, is arranged at box
The fixed seat of internal side bottom and ELISA Plate, micro pipette, centrifuge tube, colorimetric card, specification and the cover plate for being arranged at tray interior
Film, it is characterised in that：Described ELISA Plate is made up of polystyrene, using 96 hole agent plates as solid phase carrier, is put in aluminium platinum
In bag, and ELISA Plate is made up of outer frame support and the dismantled and assembled enzyme mark being placed on it reaction capillary strip, each dismantled and assembled enzyme
Mark reaction capillary strip has 12 reacting holes, thereon pre-coated emesis toxin monoclone antibody；Described micro pipette has four kinds not
The micro pipette 30 of same capacity, wherein 1.0mL capacity is used for the sampling of sample substrate, and the micro pipette 10 of 100 μ L capacity is used for
Add terminate liquid, the micro pipette 20 of 50 μ L capacity is used to adding ELIAS secondary antibody working solution, the micro pipette of 30 μ L capacity 40 with
In addition substrate developer A liquid and substrate developer B liquid；Described centrifuge tube has three kinds of different capabilities, respectively 10mL, 5mL
And 1.5mL, for sample collection and matrix pre-treatment；Described colorimetric card has 5, and display different colours reagent wells are corresponding to inhale
Shading value, the reckoning for vomitoxin residual content in water sample, dairy produce, tealeaves, feed and food；Described specification is used
In principle, standard curve, the scope of application, cross reacting rate, required instrument and reagent, application method and the attention of introducing kit
Item；Described cover plate film 5, covers ELISA Plate to prevent liquid evaporation from influenceing testing result during for sample detection；It is described
Fixed seat provided with 12 reagent bottles recessed bottle position and 2 standby recessed bottles positions of reagent bottle, wherein 12 recessed bottles of reagent bottle
Position is respectively used to place concentrated cleaning solution reagent bottle, sample diluting liquid reagent bottle, ELIAS secondary antibody working solution reagent bottle, substrate colour developing
Agent A liquid reagent bottle, substrate developer B liquid reagent bottle, terminate liquid reagent bottle, 200ng/mL vomitoxin standard solution reagent
Bottle, 100ng/mL vomitoxin standard solution reagent bottle, 50ng/mL vomitoxin standard solution reagent bottle, 25ng/
ML vomitoxin standard solution reagent bottle, 12.5ng/mL vomitoxin standard solution reagent bottle and 6.25ng/mL
Vomitoxin standard solution reagent bottle, 2 standby recessed bottle positions of reagent bottle are respectively used to place the examination of high standard product solution
Agent bottle, dilute acid soln reagent bottle, dilute alkaline soln reagent bottle or Tween-20 solution reagent bottle.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720173786.6U CN206594181U (en) | 2017-02-26 | 2017-02-26 | A kind of vomitoxin direct competive ELISA kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720173786.6U CN206594181U (en) | 2017-02-26 | 2017-02-26 | A kind of vomitoxin direct competive ELISA kit |
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| Publication Number | Publication Date |
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| CN206594181U true CN206594181U (en) | 2017-10-27 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201720173786.6U Expired - Fee Related CN206594181U (en) | 2017-02-26 | 2017-02-26 | A kind of vomitoxin direct competive ELISA kit |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110609003A (en) * | 2019-09-17 | 2019-12-24 | 苏州格锐思生物科技有限公司 | Alpha-ketoglutaric acid content determination kit and determination method thereof |
-
2017
- 2017-02-26 CN CN201720173786.6U patent/CN206594181U/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110609003A (en) * | 2019-09-17 | 2019-12-24 | 苏州格锐思生物科技有限公司 | Alpha-ketoglutaric acid content determination kit and determination method thereof |
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| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20171027 |
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| CF01 | Termination of patent right due to non-payment of annual fee |