CN206583913U - Detect the HP immunoblotting kit of common 14 food allergen IgG antibodies - Google Patents
Detect the HP immunoblotting kit of common 14 food allergen IgG antibodies Download PDFInfo
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Abstract
The utility model discloses a kind of HP immunoblotting kit for detecting common 14 food allergen IgG antibodies, it is related to vitro diagnostic techniques field.The kit, using linear immunoassay, by beef, chicken, cod, corn, crab, egg white/yolk, mushroom, milk, pork, rice, shrimp, soybean, tomato, wheat, totally 14 kinds of common food specific IgG anaphylactogens are parallel with nature controlling line and critical value line is coated on nitrocellulose filter, reacted by a clinical sample and one-time detection, it can detect the IgG antibody of 14 kinds of food allergens in human serum, overcome current Elisa methodologies kit associated antibodies and individually detect operationally cumbersome one by one, greatly improve detection efficiency.
Description
Technical field
The utility model is related to vitro diagnostic techniques field, more particularly to a kind of common 14 food allergen IgG of detection
The HP immunoblotting kit of antibody.
Background technology
At present, 14 common food anaphylactogens in clinical labororatory's detection sample, typically using ELISA
(Elisa) detected, this method is that anaphylactogen is coated in 96 microwell plates, by the incubation reaction of clinical sample, finally
Result interpretation is carried out by ELIASA.
This detection method can only detect single index due to once testing, and flux is low, and testing cost is high, for expanding
Quick former detection project has limitation.
Utility model content
The purpose of this utility model is to provide a kind of Western blotting examination for detecting common 14 food allergen IgG antibodies
Agent box, so as to solve foregoing problems present in prior art.
To achieve these goals, the technical solution adopted in the utility model is as follows：
A kind of HP immunoblotting kit for detecting common 14 food allergen IgG antibodies, including：Film bar, be arranged in parallel
14 detection lines, nature controlling line and critical value line on the film bar, 14 detection lines successively by：Beef, chicken, cod
Fish, corn, crab, egg white/yolk, mushroom, milk, pork, rice, shrimp, soybean, tomato, wheat totally 14 kinds of common food spies
Different in nature allergy primordial covering.
Preferably, in addition to transparent PVC films, the film bar is fixed on the PVC films.
Preferably, the film bar is nitrocellulose filter.
Preferably, in addition to Sample dilution, concentration washing lotion, enzyme conjugates and substrate solution, the Sample dilution be containing
Have a Tris buffer solutions of lowlenthal serum, the concentration washing lotion is 10 × Tris buffer solutions, the enzyme conjugates is alkaline phosphatase
The goat anti-human igg of enzyme mark, the substrate solution is NBT/BCIP.
The beneficial effects of the utility model are：14 common food anaphylactogens of detection that the utility model embodiment is provided
The HP immunoblotting kit of IgG antibody, using linear immunoassay, by beef, chicken, cod, corn, crab, egg white/egg
Totally 14 kinds of common food specific IgG anaphylactogens and the Quality Control of Huang, mushroom, milk, pork, rice, shrimp, soybean, tomato, wheat
Line and critical value line is parallel is coated on nitrocellulose filter, passes through a clinical sample and one-time detection is reacted, you can detection
The IgG antibody of 14 kinds of food allergens in human serum, overcomes current Elisa methodologies kit associated antibodies independent one by one
Detection operationally cumbersome, greatly improves detection efficiency.
Brief description of the drawings
Fig. 1 is the wire structures schematic diagram of the film bar for the HP immunoblotting kit that the utility model is provided；
Fig. 2 is the wiring schematic cross-section of the film bar for the HP immunoblotting kit that the utility model is provided.
In figure, the implication of each symbol is as follows：
1 nature controlling line, 2 critical value lines, 3-16 is followed successively by the detection for being coated with following 14 kinds of common food specific allergens
Line：Beef, chicken, cod, corn, crab, egg white/yolk, mushroom, milk, pork, rice, shrimp, soybean, tomato, wheat,
17 PVC films, 18 double faced adhesive tapes, 19 film bars.
Embodiment
In order that the purpose of this utility model, technical scheme and advantage are more clearly understood, below in conjunction with accompanying drawing, to this reality
It is further elaborated with new.It should be appreciated that embodiment described herein is only to explain this practicality
It is new, it is not used to limit the utility model.
As shown in Figure 1-2, the utility model embodiment provides a kind of common 14 food allergen IgG antibodies of detection
HP immunoblotting kit, including：Film bar 19, is set in parallel in 14 detection line 3-16, nature controlling line 1 and critical value on film bar 19
2,14 detection line 3-16 of line successively by：It is beef, chicken, cod, corn, crab, egg white/yolk, mushroom, milk, pork, big
Rice, shrimp, soybean, tomato, wheat totally 14 kinds of common food specific allergy primordial coverings.
In mentioned reagent box, 14 kinds of common food anaphylactogen combinations meet compatriots' eating habit, carry out 14 individually detections
It is more directly perceived accurate, provide extensive guide for the prevention and prognosis of anaphylactic disease.
The kit that the present embodiment is provided, in addition to transparent PVC films 17, film bar 19 are fixed on PVC films 17.
During actual fabrication, it can use double faced adhesive tape 18 that film bar 19 is fixed on PVC films 17.
In mentioned reagent box, film bar 19 is nitrocellulose filter.
The coating of anaphylactogen can be coated in nitrocellulose filter in high precision by automating point sample instrument, can be according to detection need
Ask progress rational sorting and combination.
The HP immunoblotting kit that the utility model embodiment is provided, in addition to Sample dilution, concentration washing lotion, enzyme are combined
Thing and substrate solution, the Sample dilution be the Tris buffer solutions containing lowlenthal serum, the concentration washing lotion be 10 × Tris
Buffer solution, the enzyme conjugates is the goat anti-human igg of alkali phosphatase enzyme mark, and the substrate solution is NBT/BCIP.
The experimental principle of mentioned reagent box is to enter the film bar for being coated with 14 kinds of common food anaphylactogens with sample buffer
Nonspecific sites in row Seal treatment, close membrane, so as to prevent the generation of nonspecific reaction.During detection, film bar and dilution
Serum sample incubate jointly and make antibody in serum and the antigen binding on film bar.Utilize alkali phosphatase enzyme mark goat-anti people
IgG detects this binding antibody.Add after substrate, the substrate for enzymatic activity reaction marked on binding antibody, and it is coated in anaphylactogen
Position produces visible band.Whether developed the color according to band, can determine whether the antibody characteristic of subject, if substantially colour developing is determined as
The antibody positive, is determined as feminine gender if not developing the color.By being coated with by carrier of nitrocellulose filter, 14 kinds are realized often
See food allergen joint-detection, substantially increase detection efficiency.
Using mentioned reagent box, 14 kinds of common food allergy can be detected simultaneously by a clinical sample and one-time detection
Original, overcomes a variety of anaphylactogens and detects operationally cumbersome one by one, greatly improve operating efficiency, be beneficial to realize high flux.
The preparation method of the HP immunoblotting kit film bar of said structure, is comprised the steps of：
1) prepare anaphylactogen coating buffer, using coating buffer solution, will totally 14 kinds of food specific allergens and anti-human igg,
Human IgG is diluted to relative coating concentration.
2) anaphylactogen prepared coating liquid spotting is coated on nitrocellulose filter using High Precision Automatic point sample instrument,
Form detection line, critical value line and nature controlling line.
3) blocking non-specific is carried out using inert protein such as bovine serum albumin(BSA), casein or NBCS.
4) dried under conditions of the film bar closed being less than into 30% as humidity.
5) dried film bar is pasted by the double faced adhesive tape of customization and be fixed on by correlated series on PVC liner plates.
6) bar assembled is cut into single part using cutting machine.
The other components of this kit include：
The HP immunoblotting kit of said structure, when in use, first carries out following pretreatment：
Concentrate the dilution of washing lotion：Before dilution it should all dissolved.Will concentration washing lotion (10 ×) and distilled water or purified water
1:9 dilutions.For example：1 part of concentration washing lotion (10 ×) plus 9 parts of distilled water or purified water.
The preparation of enzyme conjugates：10 times of concentrations.Enzyme conjugates needed for drawing, and with sample buffer 1:10 dilutions.
Its application method comprises the following steps：
1. antigenic membrane bar is placed in incubation slot, the one side with word marking is upward；1.5ml Sample Buffers are added per groove
Liquid, shaking table weak vibrations are waved 5 minutes by film bar at room temperature, and liquid in incubation slot is discarded thereafter.
2. adding the blood serum sample of 1.5ml dilutions per groove, weak vibrations are incubated 30 minutes on shaking table is waved at room temperature.
3. discarding liquid in incubation slot, 1.5ml cleaning buffer solutions are added per groove, at room temperature the weak vibrations on shaking table is waved
5 minutes, discard liquid in incubation slot.Repeat washing 2 times.
4. discarding liquid in incubation slot, the enzyme conjugates added per groove after 1.5ml dilutions is light on shaking table is waved at room temperature
It is micro- to rock incubation 30 minutes.
5. repeat step 3.
6. discarding liquid in incubation slot, 1.5ml substrate solutions are added per groove, weak vibrations are incubated on shaking table is waved at room temperature
10 minutes.
7. discarding liquid in incubation slot, 1.5ml distilled water or purified water are added per groove, it is slight on shaking table is waved at room temperature
Rock 1 minute, discard liquid in incubation slot.Repeat washing 2 times..
After 8. film bar is dried, antigenic membrane bar can be pasted on result record, to preserve experimental result.
The HP immunoblotting kit of said structure, the method for its Performance Testing is：
Conversion zone has nature controlling line, critical value line successively below detection film bar mark.If nature controlling line occurs strong
Color reaction, critical value line has faint colour developing, and nature controlling line colour developing is better than critical value line, then illustrates that experimental result is effective.Such as
Fruit nature controlling line does not occur strong color reaction, then illustrates that experimental result is invalid.
The designation strip band of result interpretation card can be alignd with the respective strap on film bar, Determination pillar location.
Can be negative and positive by result interpretation according to antigen bands colour developing degree.
It is positive：Antigen bands colour developing is consistent compared with critical value line or deeper；
It is negative：Antigen bands colour developing is shallow compared with critical value line.
By using above-mentioned technical proposal disclosed in the utility model, following beneficial effect has been obtained：The utility model
The HP immunoblotting kit for 14 common food anaphylactogen IgG antibodies of detection that embodiment is provided, using linear immunoassay,
Beef, chicken, cod, corn, crab, egg white/yolk, mushroom, milk, pork, rice, shrimp, soybean, tomato, wheat is common
14 kinds of common food specific IgG anaphylactogens are parallel with nature controlling line and critical value line to be coated on nitrocellulose filter, passes through one
Part clinical sample and one-time detection reaction, you can the IgG antibody of 14 kinds of food allergens in detection human serum, overcome current
Elisa methodology kits associated antibodies individually detect operationally cumbersome one by one, greatly improve detection efficiency.
Each embodiment in this specification is described by the way of progressive, what each embodiment was stressed be with
Between the difference of other embodiment, each embodiment identical similar part mutually referring to.
Those skilled in the art should be understood that the sequential for the method and step that above-described embodiment is provided can be entered according to actual conditions
Row accommodation, also can concurrently be carried out according to actual conditions.
All or part of step in the method that above-described embodiment is related to can be instructed by program correlation hardware come
Complete, described program can be stored in the storage medium that computer equipment can be read, for performing the various embodiments described above side
All or part of step described in method.The computer equipment, for example：Personal computer, server, the network equipment, intelligent sliding
Dynamic terminal, intelligent home device, wearable intelligent equipment, vehicle intelligent equipment etc.；Described storage medium, for example：RAM、
ROM, magnetic disc, tape, CD, flash memory, USB flash disk, mobile hard disk, storage card, memory stick, webserver storage, network cloud storage
Deng.
Finally, in addition it is also necessary to explanation, herein, such as first and second or the like relational terms be used merely to by
One entity or operation make a distinction with another entity or operation, and not necessarily require or imply these entities or operation
Between there is any this actual relation or order.Moreover, term " comprising ", "comprising" or its any other variant meaning
Covering including for nonexcludability, so that process, method, commodity or equipment including a series of key elements not only include that
A little key elements, but also other key elements including being not expressly set out, or also include be this process, method, commodity or
The intrinsic key element of equipment.In the absence of more restrictions, the key element limited by sentence "including a ...", is not arranged
Except also there is other identical element in the process including the key element, method, commodity or equipment.
Described above is only preferred embodiment of the present utility model, it is noted that for the common skill of the art
For art personnel, on the premise of the utility model principle is not departed from, some improvements and modifications can also be made, these improve and
Retouching should also regard protection domain of the present utility model.
Claims (1)
1. a kind of HP immunoblotting kit for detecting common 14 food allergen IgG antibodies, it is characterised in that including：Film bar,
Be set in parallel in 14 detection lines, nature controlling line and critical value line on the film bar, 14 detection lines successively by：Beef,
Chicken, cod, corn, crab, egg white/yolk, mushroom, milk, pork, rice, shrimp, soybean, tomato, wheat totally 14 kinds it is common
Food specific allergy primordial covering；Also include Sample dilution, concentration washing lotion, enzyme conjugates and substrate solution, the Sample Dilution
Liquid be the Tris buffer solutions containing lowlenthal serum, the concentration washing lotion be 10 × Tris buffer solutions, the enzyme conjugates be alkali
The goat anti-human igg of acid phosphatase mark, the substrate solution is NBT/BCIP；
The film bar, is fixed on described by described HP immunoblotting kit, in addition to transparent PVC films using double faced adhesive tape
On PVC films；
The film bar is nitrocellulose filter.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201621035273.0U CN206583913U (en) | 2016-08-31 | 2016-08-31 | Detect the HP immunoblotting kit of common 14 food allergen IgG antibodies |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201621035273.0U CN206583913U (en) | 2016-08-31 | 2016-08-31 | Detect the HP immunoblotting kit of common 14 food allergen IgG antibodies |
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| CN206583913U true CN206583913U (en) | 2017-10-24 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115792248A (en) * | 2023-02-13 | 2023-03-14 | 江西赛基生物技术有限公司 | Food-specific IgG antibody detection kit and use method thereof |
-
2016
- 2016-08-31 CN CN201621035273.0U patent/CN206583913U/en active Active
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115792248A (en) * | 2023-02-13 | 2023-03-14 | 江西赛基生物技术有限公司 | Food-specific IgG antibody detection kit and use method thereof |
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