CN206546369U - The HP immunoblotting kit of the comprehensive 42 food allergen IgG antibodies of detection - Google Patents

Abstract

The utility model discloses a kind of HP immunoblotting kit for detecting comprehensive 42 food allergen IgG antibodies, it is related to vitro diagnostic techniques field.The kit, using linear immunoassay, by beef, chicken, cod, corn, crab, egg, mushroom, milk, pork, rice, shrimp, soybean, tomato, wheat, watermelon, olive, blueberry, pineapple, orange, peach, apple, durian, mango, banana, "Hami" melon, grape, shaddock, strawberry, lobster, salmon, sardine, grass carp, hairtail, scallop, clam, sesame, spinach, green soya bean, turkey, mutton, cashew nut, totally 42 kinds of common food specific IgG anaphylactogens are parallel with nature controlling line and critical value line is coated on nitrocellulose filter for peanut, reacted by a clinical sample and one-time detection, it can detect the IgG antibody of 42 kinds of food allergens in human serum, overcome existing antibody and individually detect operationally cumbersome one by one, greatly improve detection efficiency.

Description

The HP immunoblotting kit of the comprehensive 42 food allergen IgG antibodies of detection

Technical field

The utility model is related to vitro diagnostic techniques field, more particularly to a kind of comprehensive 42 food allergen IgG of detection The HP immunoblotting kit of antibody.

Background technology

At present, 42 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 comprehensive 42 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 comprehensive 42 food allergen IgG antibodies, including：Film bar, be arranged in parallel 42 detection lines, nature controlling line and critical value line on the film bar, 42 detection lines successively by：Beef, chicken, cod Fish, corn, crab, egg, mushroom, milk, pork, rice, shrimp, soybean, tomato, wheat, watermelon, olive, blueberry, spinach Trailing plants, orange, peach, apple, durian, mango, banana, "Hami" melon, grape, shaddock, strawberry, lobster, salmon, sardine, grass carp, Hairtail, scallop, clam, sesame, spinach, green soya bean, turkey, mutton, cashew nut, peanut totally 42 kinds of common food specific allergen bags Quilt.

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：42 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, mushroom Mushroom, milk, pork, rice, shrimp, soybean, tomato, wheat, watermelon, olive, blueberry, pineapple, orange, peach, apple, durian, Mango, banana, "Hami" melon, grape, shaddock, strawberry, lobster, salmon, sardine, grass carp, hairtail, scallop, clam, sesame, spinach Totally 42 kinds of common food specific IgG anaphylactogens and nature controlling line and critical value line are flat for dish, green soya bean, turkey, mutton, cashew nut, peanut Row is coated on nitrocellulose filter, is reacted by a clinical sample and one-time detection, you can 42 kinds in detection human serum The IgG antibody of food allergen, overcomes current Elisa methodologies kit associated antibodies and individually detects one by one operationally It is cumbersome, greatly improve 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, 3-44 is followed successively by the detection for being coated with following 42 kinds of common food specific allergens Line：Beef, chicken, cod, corn, crab, egg, mushroom, milk, pork, rice, shrimp, soybean, tomato, wheat, west Melon, olive, blueberry, pineapple, orange, peach, apple, durian, mango, banana, "Hami" melon, grape, shaddock, strawberry, lobster, three texts Fish, sardine, grass carp, hairtail, scallop, clam, sesame, spinach, green soya bean, turkey, mutton, cashew nut, peanut, 45PVC films, 46 pairs Face glue, 47 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 comprehensive 42 food allergen IgG antibodies of detection HP immunoblotting kit, including：Film bar 47, is set in parallel in 42 detection line 3-44 on the film bar, nature controlling line 1 and critical Be worth line 2,42 detection line 3-44 successively by：Beef, chicken, cod, corn, crab, egg, mushroom, milk, pork, rice, Shrimp, soybean, tomato, wheat, watermelon, olive, blueberry, pineapple, orange, peach, apple, durian, mango, banana, "Hami" melon, Portugal Grape, shaddock, strawberry, lobster, salmon, sardine, grass carp, hairtail, scallop, clam, sesame, spinach, green soya bean, turkey, mutton, waist Really, peanut totally 42 kinds of common food specific allergy primordial coverings.

In mentioned reagent box, the combination of 42 kinds of common food anaphylactogens meets compatriots' eating habit, carries out 42 comprehensive inspections Survey more directly perceived accurate, extensive guide is provided for the prevention and prognosis of anaphylactic disease.

The kit that the present embodiment is provided, in addition to transparent PVC films 45, film bar 47 are fixed on PVC films 45.

During actual fabrication, it can use double faced adhesive tape 46 that film bar 47 is fixed on PVC films 45.

The kit that the present embodiment is provided, film bar 47 is nitrocellulose filter.

The coating of anaphylactogen is coated in nitrocellulose filter in high precision by automating point sample instrument, can be entered according to detection demand Row rational sorting and combination.

The kit that the present embodiment is provided, in addition to Sample dilution, concentration washing lotion, enzyme conjugates and substrate solution, it is described Sample dilution be the Tris buffer solutions containing lowlenthal serum, the concentration washing lotion be 10 × Tris buffer solutions, the enzyme knot Compound 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 42 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, 42 kinds are realized often See food allergen joint-detection, substantially increase detection efficiency.

Using mentioned reagent box, 42 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 HP immunoblotting kit of said structure, the preparation method of film bar, is comprised the steps of：

1) prepare anaphylactogen coating buffer, using coating buffer solution, will totally 42 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, carries out following pretreatment first：

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 42 common food anaphylactogen IgG antibodies of detection that embodiment is provided, using linear immunoassay, By beef, chicken, cod, corn, crab, egg, mushroom, milk, pork, rice, shrimp, soybean, tomato, wheat, watermelon, Olive, blueberry, pineapple, orange, peach, apple, durian, mango, banana, "Hami" melon, grape, shaddock, strawberry, lobster, salmon, Totally 42 kinds of common foods are special for sardine, grass carp, hairtail, scallop, clam, sesame, spinach, green soya bean, turkey, mutton, cashew nut, peanut IgG anaphylactogens are parallel with nature controlling line and critical value line is coated on nitrocellulose filter for property, by a clinical sample and once Detection reaction, you can the IgG antibody of 42 kinds of food allergens in detection human serum, overcomes current Elisa methodologies reagent Box 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 comprehensive 42 food allergen IgG antibodies, it is characterised in that including：Film bar, Be set in parallel in 42 detection lines, nature controlling line and critical value line on the film bar, 42 detection lines successively by：Beef, Chicken, cod, corn, crab, egg, mushroom, milk, pork, rice, shrimp, soybean, tomato, wheat, watermelon, olive, indigo plant The certain kind of berries, pineapple, orange, peach, apple, durian, mango, banana, "Hami" melon, grape, shaddock, strawberry, lobster, salmon, sardine, Grass carp, hairtail, scallop, clam, sesame, spinach, green soya bean, turkey, mutton, cashew nut, peanut totally 42 kinds of common food specific allergies Primordial covering；Also include Sample dilution, concentration washing lotion, enzyme conjugates and substrate solution, the Sample dilution is to contain blood of goats Clear Tris buffer solutions, the concentration washing lotion is 10 × Tris buffer solutions, the enzyme conjugates is alkali phosphatase enzyme mark Goat anti-human igg, 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.