CN206710440U - The fluorescence immune chromatography kit of Quantitative detection vomitoxin - Google Patents
The fluorescence immune chromatography kit of Quantitative detection vomitoxin Download PDFInfo
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- CN206710440U CN206710440U CN201720357308.0U CN201720357308U CN206710440U CN 206710440 U CN206710440 U CN 206710440U CN 201720357308 U CN201720357308 U CN 201720357308U CN 206710440 U CN206710440 U CN 206710440U
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Abstract
The utility model belongs to technical field of biological, more particularly to a kind of fluorescence immune chromatography kit of Quantitative detection vomitoxin, it is described including box body and lid, side sealed box body of the lid from box opening, lid is provided with well and observation window, what the inwall of lid slided is provided with seal, well is opened or closed in seal sliding process, immuno-chromatographic test paper strip is provided with box body, the immuno-chromatographic test paper strip includes bottom plate, on bottom plate sample pad is overlapped with according to by the square upstream sequence of one end to the other end, pad, reacting pad and adsorptive pads, fluorescent microsphere mark vomitoxin antibody and fluorescent microsphere mark chicken IgY are adsorbed with pad, reacting pad is provided with detection zone and quality control region, vomitoxin antigen protein is coated with detection zone, goat-anti chicken IgY is coated with quality control region.Kit of the present utility model can fast and efficiently detect vomitoxin.
Description
Technical field
The utility model belongs to technical field of biological, and in particular to a kind of fluorescence of Quantitative detection vomitoxin
Immune chromatography reagent kit.
Background technology
Vomitoxin bulk composition is DON (deoxynivalenol, deoxynivalenol), and it is mould to belong to single-ended spore
Aliphatic compound, mainly by Fusarium graminearum, Fusarium oxysporum, fusarium moniliforme, Fusarium sporotrichioides, Fusarlum roseum, snow
The sickle-like bacteria such as rotten sickle-like bacteria produce.In addition, the bacterial strain of Cephalosporium sp, paint class Pseudomonas, trichoderma etc. can all produce the toxin.It is single-ended
Bao Mei alkene race toxin shares kind more than 150, is a kind of strong immunodepressant, and caused classical symptom is that feed intake reduces, institute
Feed food refusal toxin is called with this toxoid.Vomitoxin (DON) is a kind of wherein most important toxin, essentially from Fusarium
(Fusarium), especially Fusarium graminearum (Fusariumgraminearum) and fusarium culmorum (Fusarium
culmorum).Because it can cause the vomiting of pig, therefore also known as vomitoxin (vomitoxin, VT).
Vomitoxin is mycotoxin common in food, is widespread in nature, generally in Europe and North America hair
3 kinds of existing toxin are deoxynivalenol, 3- acetyl deoxynivalenol and 15- deoxynivalenol bacterium alkene
Alcohol.Because they have very high cytotoxin and immunosupress property, therefore, prestige is constituted to the health of the mankind and animal
The side of body, particularly there is obvious influence on immunologic function.According to DON dosage and open-assembly time difference can cause immunosupress or
Immunostimulation.After people has taken in the food by DON pollutions, apocleisis, vomiting, diarrhoea, fever, astasia, reaction can be caused
The acute poisoning symptom such as blunt, hemopoietic system is damaged when serious and is caused death.Due to Cereals ratio in Chinese tradition eating habit
It is much higher than west so that the harm of vomitoxin is more prominent.1998, in the evaluation report that international cancer research institution announces
In announcement, vomitoxin is listed in 3 class carcinogenic substances.China's cereal, swine feed, calf mixed feed, lactation period animal coordinate
DON limit standard is 1.0mg/kg in feed, and DON limit standard is 5.0mg/ in ox mixed feed, poultry mixed feed
kg。
The method of detection vomitoxin has a variety of at present, such as thin-layered chromatography, EUSA (ELISA), gas
Phase chromatogram, high performance liquid chromatography, infrared spectrum analysis etc..Efficient liquid chromatography mutually belongs to National Standard Method, and high sensitivity, the degree of accuracy are high,
But expensive instrument, the complex pretreatment of sample, cost time length are needed, testing cost is high, it is impossible to execute-in-place, and need special
Industry human users, so limiting its application.Enzyme-linked immunosorbent assay is high to detection environmental requirement, it is necessary to which professional grasps
Make, be adapted to the detection of great amount of samples, and have corresponding data processing.The fluorogenic quantitative detection test strips of vomitoxin coordinate phase
The Fluorescent reader answered, has that the degree of accuracy is high, and reproducible, testing cost is low, and environmental requirement is low, simple to operate, just with data
Management, preserve and upload the characteristics of.This product be applied to Quantitative detection foodstuff grain (rice, corn, wheat, barley,
Sorghum etc.) and its product, feed and its raw material in vomitoxin residual concentration, sample pre-treatments are simple, whole detection process
18min or so (pre-treatment 8min+ detects 8min) is only needed, the CUT OFF values of vomitoxin fluorogenic quantitative detection test strips are
1000ppb, sensitivity 100ppb, TIANZHU XINGNAO Capsul 80%~125%, the range of linearity are 100ppb~5000ppb,
Under 10000ppb concentration with other toxin no cross reactions.Suitable for all kinds of foodstuff grain processing enterprises, third party's detection machine
Structure and supervision department of governments at all levels.
Utility model content
The purpose of this utility model is to provide a kind of fluorescence immune chromatography kit of Quantitative detection vomitoxin, can
Fast and efficiently detect vomitoxin.
To achieve the above object, technical scheme is used by the utility model：Including box body and lid, lid is from box body
The side sealed box body of opening, lid are provided with well and observation window, and the inwall slip of lid is provided with seal, seal
Well is opened or closed in sliding process, is provided with immuno-chromatographic test paper strip in box body, the immuno-chromatographic test paper strip includes bottom
Plate, sample pad, pad, reacting pad and adsorptive pads are overlapped with according to by the square upstream sequence of one end to the other end on bottom plate, tie
Close pad on is adsorbed with fluorescent microsphere mark vomitoxin antibody and fluorescent microsphere mark chicken IgY, reacting pad be provided with detection zone and
Quality control region, vomitoxin antigen protein is coated with detection zone, goat-anti chicken IgY is coated with quality control region.
Preferably, the fluorescent microsphere is Eu3+The latex beads of mark.
Preferably, the reacting pad is polyester cellulose film.
Preferably, the inwall of the lid is provided with chute, and the both sides of seal are in chute.
Preferably, the seal is connected with a slider, and lid is provided with strip hole, and strip hole is parallel with chute, slider
Stretched out through strip hole and from the top surface of lid.
Preferably, the bottom of the box body is provided with keeper, and it is spacing that the immuno-chromatographic test paper strip is positioned part fixation.
Preferably, the keeper is a snap ring.
Preferably, one end of the box body and lid is hinged, and the other end makes box body be closed with lid by way of snapping.
After adopting the above technical scheme, the utility model has the positive effect that：
(1) immuno-chromatographic test paper strip in the utility model, observe by the naked eye and can determine whether vomitoxin yin and yang attribute, adopt
The fluorescence intensity of T lines and C lines is read with Fluorescent reader and calculates T/C values, can be quantified by the standard curve built in instrument
The content for calculating vomitoxin in sample, meanwhile, quantitative testing result can be presented on Fluorescent reader LCDs,
The printable examining report for obtaining papery simultaneously；
(2) immuno-chromatographic test paper strip is positioned in kit, and well is covered by seal, can store for a long time
Test strips, test strips are prevented to be contaminated.When needing detection, directly it is loaded from well, qualitatively judges negative and positive from observation window
Property, if need to be quantified to vomitoxin, test strips can be taken out from kit, be put into the mould supporting with fluorescence detector
In tool, or directly kit can be put into fluorescence detector and detected,.
Brief description of the drawings
Fig. 1 is the structure chart of immuno-chromatographic test paper strip in the utility model；
Fig. 2 is the top view of immuno-chromatographic test paper strip shown in Fig. 1；
Fig. 3 is the structure chart of the fluorescence immune chromatography kit of Quantitative detection vomitoxin of the present utility model；
Fig. 4 is the structure chart for removing box body rear cover body；
Fig. 5 is the structure chart of box body after removal lid.
Wherein：1st, bottom plate, 2, sample pad, 3, pad, 4, reacting pad, 401, detection zone, 402, quality control region, 5, water suction
Pad, 6, box body, 7, lid, 8, well, 9, observation window, 10, chute, 11, seal, 12, slider, 13, strip hole, 14, fixed
Position part.
Embodiment
The utility model is described in further detail with reference to the accompanying drawings and detailed description.
In the utility model, unless otherwise clearly defined and limited, term " installation ", " connected ", " connection ", " Gu
It is fixed " etc. term should broadly understood, for example, it may be fixedly connected or be detachably connected, or integrally；Can be
Mechanically connect or electrically connect；Can be joined directly together, can also be indirectly connected by intermediary, can be two
The connection of element internal or the interaction relationship of two elements.For the ordinary skill in the art, can basis
Concrete condition understands concrete meaning of the above-mentioned term in the utility model.
1. as shown in figure 1, be the immuno-chromatographic test paper strip in the utility model, including bottom plate 1, according to by one on bottom plate 1
Hold the square upstream sequence to the other end to be overlapped with sample pad 2, pad 3, reacting pad 4 and adsorptive pads 5, be adsorbed with pad 3
Fluorescent microsphere marks vomitoxin antibody and fluorescent microsphere mark chicken IgY, and reacting pad 4 is provided with detection zone 401 and quality control region
402, vomitoxin antigen protein is coated with detection zone 401, and goat-anti chicken IgY is coated with quality control region 402.
Wherein fluorescent microsphere mark vomitoxin antibody is to be coupled to obtain by vomitoxin antibody and latex beads, and fluorescence is micro-
Ball mark chicken IgY is to be coupled to obtain by chicken IgY and latex beads.
2. a kind of fluorescence immune chromatography kit of Quantitative detection vomitoxin, including box body 6 and lid 7, lid 7
One end of the side sealed box body 6 being open from box body 6, box body 6 and lid 7 is hinged, and the other end makes box body 6 by way of snapping
Closed with lid 7.
Lid 7 is provided with well 8 and observation window 9, and the inwall of lid 7 is provided with seal by what a chute 10 slided
11, seal 11 is connected with a slider 12, and lid 7 is provided with strip hole 13, and strip hole 13 is parallel with chute 10, and slider 12 passes through
Strip hole 13 is simultaneously stretched out from the top surface of lid 7, and well 8 can be opened or close in the sliding process of seal 11.
The bottom of box body 6 is provided with keeper 14, the immuno-chromatographic test paper strip being provided with box body 6 in this case, the immune layer
Analysis test strips be positioned part 14 fix it is spacing.Wherein, the keeper 14 is a snap ring, and test strips are blocked.
3. the preparation method of this case immuno-chromatographic test paper strip is：
(1) will be without any carrier base phenylethylene micro ball parcel Eu3+It is micro- to obtain latex synthetic, with label
Ball；
(2) vomitoxin antibody and chicken IgY are coupled with the latex beads with label respectively；
(3) vomitoxin antibody labeling thing and chicken IgY labels are uniformly sprayed on pad 3,37 degrees Celsius of drying
24h, latex mattress is obtained, is saved backup in a dry environment；
(4) vomitoxin antigen protein is drawn on polyester cellulose film at detection zone 401, goat-anti chicken IgY is drawn in polyester
On cellulose membrane at quality control region 402,37 degrees Celsius of drying 24h, reacting pad 4 is obtained, is saved backup in a dry environment；
(5) paste sample pad 2, pad 3, reacting pad 4, adsorptive pads 5 in order on bottom plate 1, it is wide to be finally cut into 4mm
Small bar, be fitted into kit, can be preserved 12 months under the conditions of 4~30 DEG C.
4. application this case in kit, quantitatively detect foodstuff grain feed in vomitoxin remain method, it include with
Lower step：
1) processing of sample
2) dilution of sample
3) it is loaded
4) reading
Above-mentioned immuno-chromatographic test paper strip uses the lateral immunochromatography of fluorescence (Fluorescent Lateral Flow, FLF)
Methodology principle.When testing sample is added dropwise in well 8, the vomitoxin in sample and the fluorescent microsphere in pad 3
Mark vomitoxin antibody binding is simultaneously chromatographed forward by capillarity, after reaching detection zone 401, detects fixed on T lines vomit
Tell Venom antigens and remaining uncombined fluorescent microsphere mark vomitoxin antibody binding.The fluorescence combined on detection line T lines is micro-
The concentration of vomitoxin is inversely proportional in the amount and sample of ball mark vomitoxin antibody, and nature controlling line C line combines in quality control region 402
Fluorescent marker is unrelated with the concentration of vomitoxin in sample.After chromatography terminates, T lines and C lines are read using Fluorescent reader
Fluorescence intensity simultaneously calculates T/C values, can calculate the content of vomitoxin in sample by the standard curve built in instrument and sentence
Disconnected yin and yang attribute.Quantitative testing result will be presented on Fluorescent reader LCDs, while can be printed by printing key and be obtained paper
The examining report of matter, in addition, after opening the WIFI data upload functions of instrument, detection related data information will be uploaded to automatically
" food security trace to the source management cloud platform ", is easy to trace to the source and quality management.
Negative (-)：Testing result ＜ cutoff values (can be adjusted) according to the internal control quality standard of client, be as a result the moon
Property；
Positive (+)：Testing result >=cutoff values (can be adjusted) according to the internal control quality standard standard of client, as a result for
It is positive；
It is invalid：Readout instrument reports an error, then this detection is invalid, need to retest；
If testing result exceeds the upper limit of the range of linearity, the supernatant after sample extraction centrifugation is can use, uses sample extraction
Liquid detects (taking 100 μ l supernatants, add 400 μ l extract solutions) again after diluting 5 times, and testing result is multiplied by 5, and as sample is final
Concentration.
5. pattern detection
1) blank grain trough sample is taken, corn, rice, wheat, each portion of flour, weighs 7 parts respectively, addition vomiting poison
Plain standard items final concentration is to 0ppb, 100ppb, 250ppb, 500ppb, 1000ppb, 2500ppb, 5000ppb.Used after extraction
Kit detects.Ten Duplicate Samples of each pattern detection, it is as shown in table 1 to read data.
The sample of table 1 adds testing result
The market sample chromatogram testing result of table 2 and vomitoxin fluorogenic quantitative detection ELISA test strip Comparative result
Market sample chromatogram detected value and vomitoxin fluorogenic quantitative detection ELISA test strip value coincidence rate, linearity curve
For：Y=0.986x+24.1, R2=0.989.
As a result show：With vomitoxin fluorogenic quantitative detection ELISA test strip foodstuff grain feed sample, and efficient liquid phase
Chromatogram value compares, and coincidence rate reaches 98.9%, accuracy range 80%~125%, CV≤15%.Meet Quantitative detection
It is required that.
2) specific detection
With vomitoxin fluorogenic quantitative detection test strips, 10ug/ml zearalenone is detected, aflatoxin B1,
Ochratoxin A, fumonisins, T-2 toxin, as a result display are respectively less than 100ng/ml.Illustrate vomitoxin fluorogenic quantitative detection
Test strips, with zearalenone, aflatoxin B1, ochratoxin A, fumonisins, the equal no cross reaction of T-2 toxin.
It will be apparent to those skilled in the art that technical scheme that can be as described above and design, make other various
Corresponding change and deformation, and all these changes and deformation should all belong to the protection domain of the requires of the utility model
Within.
Claims (8)
- A kind of 1. fluorescence immune chromatography kit of Quantitative detection vomitoxin, it is characterised in that：Including box body (6) and lid Body (7), the side sealed box body (6) that lid (7) is open from box body (6), lid (7) are provided with well (8) and observation window (9) what, the inwall of lid (7) slided is provided with seal (11), opens or close well in seal (11) sliding process (8), be provided with immuno-chromatographic test paper strip in box body (6), the immuno-chromatographic test paper strip includes bottom plate (1), on bottom plate (1) according to by One end to the square upstream sequence of the other end is overlapped with sample pad (2), pad (3), reacting pad (4) and adsorptive pads (5), pad (3) fluorescent microsphere mark vomitoxin antibody and fluorescent microsphere mark chicken IgY are adsorbed with, reacting pad (4) is provided with detection zone (401) vomitoxin antigen protein and quality control region (402), is coated with detection zone (401), quality control region is coated with sheep on (402) Anti- chicken IgY.
- 2. the fluorescence immune chromatography kit of Quantitative detection vomitoxin according to claim 1, it is characterised in that： The fluorescent microsphere is Eu3+The latex beads of mark.
- 3. the fluorescence immune chromatography kit of Quantitative detection vomitoxin according to claim 1, it is characterised in that： Reacting pad (4) is polyester cellulose film.
- 4. the fluorescence immune chromatography kit of Quantitative detection vomitoxin according to claim 1, it is characterised in that： The inwall of lid (7) is provided with chute (10), and the both sides of seal (11) are in chute (10).
- 5. the fluorescence immune chromatography kit of Quantitative detection vomitoxin according to claim 4, it is characterised in that： Seal (11) is connected with a slider (12), and lid (7) is provided with strip hole (13), and strip hole (13) is parallel with chute (10), Slider (12) stretches out through strip hole (13) and from the top surface of lid (7).
- 6. the fluorescence immune chromatography kit of Quantitative detection vomitoxin according to claim 1, it is characterised in that： The bottom of box body (6) is provided with keeper (14), and it is spacing that the immuno-chromatographic test paper strip is positioned part (14) fixation.
- 7. the fluorescence immune chromatography kit of Quantitative detection vomitoxin according to claim 6, it is characterised in that： The keeper (14) is a snap ring.
- 8. the fluorescence immune chromatography kit of Quantitative detection vomitoxin according to claim 1, it is characterised in that： One end of box body (6) and lid (7) is hinged, and the other end makes box body (6) be closed with lid (7) by way of snapping.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201720357308.0U CN206710440U (en) | 2017-04-07 | 2017-04-07 | The fluorescence immune chromatography kit of Quantitative detection vomitoxin |
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| CN201720357308.0U CN206710440U (en) | 2017-04-07 | 2017-04-07 | The fluorescence immune chromatography kit of Quantitative detection vomitoxin |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110133264A (en) * | 2019-06-11 | 2019-08-16 | 上海雄图生物科技有限公司 | A kind of time-resolved fluoroimmunoassay chromatography kit detecting Mycotoxins in Feed |
| CN110133260A (en) * | 2019-06-11 | 2019-08-16 | 上海雄图生物科技有限公司 | A kind of time-resolved fluoroimmunoassay chromatography kit detecting meat, eggs and milk veterinary drug residue |
| CN111487230A (en) * | 2020-04-20 | 2020-08-04 | 中国农业科学院烟草研究所 | Ralstonia solanacearum detection device |
-
2017
- 2017-04-07 CN CN201720357308.0U patent/CN206710440U/en active Active
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110133264A (en) * | 2019-06-11 | 2019-08-16 | 上海雄图生物科技有限公司 | A kind of time-resolved fluoroimmunoassay chromatography kit detecting Mycotoxins in Feed |
| CN110133260A (en) * | 2019-06-11 | 2019-08-16 | 上海雄图生物科技有限公司 | A kind of time-resolved fluoroimmunoassay chromatography kit detecting meat, eggs and milk veterinary drug residue |
| CN111487230A (en) * | 2020-04-20 | 2020-08-04 | 中国农业科学院烟草研究所 | Ralstonia solanacearum detection device |
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