CN202433382U - Solid phase carrier for enzyme-linked immunosorbent assay and kit - Google Patents
Solid phase carrier for enzyme-linked immunosorbent assay and kit Download PDFInfo
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- CN202433382U CN202433382U CN2011205430239U CN201120543023U CN202433382U CN 202433382 U CN202433382 U CN 202433382U CN 2011205430239 U CN2011205430239 U CN 2011205430239U CN 201120543023 U CN201120543023 U CN 201120543023U CN 202433382 U CN202433382 U CN 202433382U
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- solid phase
- phase carrier
- linked immunosorbent
- immunosorbent assay
- enzyme
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Abstract
The utility model discloses a solid phase carrier for enzyme-linked immunosorbent assay and a kit containing the solid phase carrier. According to the solid phase carrier, a cellulose membrane which can be combined with protein, particularly a nitrocellulose membrane is adopted to replace the conventional polystyrene porous plate, so that detection sensitivity can be improved, and the using amount of enzyme markers and substrates can be reduced, and thus, the cost is saved, and the price of products is reduced.
Description
Technical field
The utility model relates to the enzyme linked immunosorbent assay (ELISA) field, particularly, relates to a kind of solid phase carrier and kit that is used for enzyme linked immunosorbent assay (ELISA).
Background technology
Sweden scholar Engvail in 1971 and Perlmann; Holland scholar Van Weerman and Schuurs report respectively immunological technique are developed into the solid-phase immunoassay method that detects micro substance in the body fluid; Be enzyme-linked immunosorbent assay (enzyme-linked immunosorbent assay, ELISA).ELISA has become the advanced subject in the present analytical chemistry field now, and it is a kind of special reagent analysis method, is a kind of novel immunoassay that on the basis of immunoenzyme technics (immunoenzymatic techniques), grows up.
Ultimate principle:
It adopts antigen with the idiosyncrasy of antibody determinand to be connected with enzyme, produces color reaction through enzyme-to-substrate then, is used for quantitative measurement.The object of measuring can be that antibody also can be antigen.
The reagent that 3 kinds of necessity are arranged in this assay method:
1. the antigen of solid phase or antibody (immunosorbent)
2. the antigen of enzyme labeling or antibody (label)
3. the substrate of enzyme effect (developer)
During measurement; Antigen (antibody) is combined on the solid phase carrier earlier, but still keeps its immunocompetence, adds the conjugate (label) that a kind of antibody (antigen) and enzyme are combined into then; This conjugate still keeps its former immunocompetence and enzymatic activity; After the antigen on conjugate and the solid phase carrier (antibody) reaction bonded, add the corresponding substrate of enzyme, promptly play catalyzing hydrolysis or redox reaction and be color.The shade that it generated is directly proportional with antigen (antibody) content that desire is surveyed.This coloured product can be with the naked eye, optical microscope, electron microscope observation, also can use spectrophotometer (ELIASA) to measure.Its method is simple, convenient news speed, high specificity.
ELISA kit of the prior art generally adopts polystyrene micropore plate (40 or 96 hole) as solid phase carrier; With specific antigen or antibody sandwich in the micropore of polystyrene micropore plate; Because this solid phase carrier is to exist with the porous mode, when detecting, need to add enzyme labeling thing and substrate by the hole; Cause the use amount of enzyme labeling thing and substrate big, cost of products is high, sensitivity is low.
The utility model content
The purpose of the utility model is to overcome the shortcoming and defect of above-mentioned prior art; A kind of solid phase carrier that is used for enzyme linked immunosorbent assay (ELISA) is provided, can reduces the use amount of enzyme labeling thing and substrate, thereby practice thrift cost; Reduce product price, and the sensitivity that can improve mensuration.
Another purpose of the utility model is to provide a kind of kit that is used for enzyme linked immunosorbent assay (ELISA).
The technical scheme that realizes above-mentioned purpose is following:
A kind of solid phase carrier that is used for enzyme linked immunosorbent assay (ELISA), said solid phase carrier are can protein-bonded cellulose membrane.
Further, said cellulose membrane is a nitrocellulose filter.
Further, on the said nitrocellulose filter detection line can be set, said detection line position encapsulate can with the specificity anaphylactogen of IgE antibodies in the blood.
Further, also can be provided with nature controlling line on the said nitrocellulose filter.
A kind of kit that is used for enzyme linked immunosorbent assay (ELISA) comprises above-mentioned solid phase carrier.
Owing to adopt technique scheme, the utlity model has following beneficial effect:
1, adopt can protein-bonded cellulose membrane, especially nitrocellulose filter for the utility model, replaces traditional polystyrene micropore plate, can reduce the use amount of enzyme labeling thing and substrate, thus the saving cost, the reduction product price.
2, the utility model adopts nitrocellulose filter as solid phase carrier; Can improve detection sensitivity; Even the specific IgE antibody that the content in blood sample is extremely low through specific allergen being combined in the detection line position of nitrocellulose filter, also can detect efficiently.
3, nature controlling line being set on nitrocellulose filter, whether correct or whether reagent invalid, improve to be detected as power if being used for detecting operation.
Through accompanying drawing and embodiment, the technical scheme of the utility model is done further detailed description below.
Description of drawings
Fig. 1 is the solid phase carrier structural representation that is used for enzyme linked immunosorbent assay (ELISA) of the utility model.
Embodiment
Describe below in conjunction with the preferred embodiment of accompanying drawing, should be appreciated that preferred embodiment described herein only is used for explanation and explains the utility model, and be not used in qualification the utility model the utility model.
Present embodiment is that example describes with the extremely low specific IgE antibody of content that detects in the blood sample.
As shown in Figure 1; The solid phase carrier that is used for enzyme linked immunosorbent assay (ELISA) of the utility model is for can protein-bonded cellulose membrane 1; Preferred nitrocellulose filter; Detection line 2 and nature controlling line 3 can be set on the nitrocellulose filter, detection line 2 positions encapsulate can with the specificity anaphylactogen (the spray film encapsulates processing) of IgE antibodies in the blood.
Wherein the disposal route of anaphylactogen spray nitrocellulose filter is following:
(1) anaphylactogen is dissolved in the PBS (PH:8.0) of 100mM, makes its concentration reach 1mg/ml.
(2) BioDot Membrane jetter parameter is set to Rate value 1.0ul/cm.
(3) nitrocellulose filter is lain on the operator's console, in humidity 30%-50% scope, spray film.
(4) place 1-2 hour at ambient temperature, make its drying.
The spray film method of nature controlling line is the same.
The kit that is used for enzyme linked immunosorbent assay (ELISA) of the utility model comprises above-mentioned solid phase carrier (containing detection line and nature controlling line) and cleaning fluid (PBST), ELIAS secondary antibody (the mouse-anti human IgE monoclonal antibody of horseradish peroxidase-labeled) and substrate (tetramethyl benzidine TMB), stop buffer.
The process of utilizing the mentioned reagent box to detect is following:
The nitrocellulose filter that (1) will spray anaphylactogen places incubation slot, adds the 1ml blood serum sample, hatches 30 minutes waving to rock on the shaking table under the room temperature.
(2) liquid in the incubation slot is removed in suction.Every groove adds the 1.5ml cleaning fluid, rocks 5 minutes on the shaking table waving under the room temperature, inhales and removes liquid in the incubation slot.Repeated washing is 2 times again.
(3) liquid in the incubation slot is removed in suction.Every groove adds the 1ml ELIAS secondary antibody, hatches 30 minutes waving to rock on the shaking table under the room temperature.
(4) repeating step (2) washing.
(5) liquid in the incubation slot is removed in suction.Every groove adds the 1ml substrate, hatches 10 minutes waving slightly to rock on the shaking table under the room temperature.
(6) liquid in the incubation slot is removed in suction.Every groove adds 1.5ml distilled water, rocks washing 1 minute under the room temperature.
(7) liquid in the incubation slot is removed in suction.Add the 1ml stop buffer in every groove, rock under the room temperature and hatch 5 minutes.
(8) nitrocellulose filter is placed drying on the filter paper, test strip (detection line district) and positive band (nature controlling line district) are compared judged result.
The result explains
Positive: respectively in the detection line district and the nature controlling line district a blue band respectively appears;
Negative: as a blue ribbon only to occur in the nature controlling line district;
Blue ribbon does not appear in the nature controlling line district, shows that incorrect operation or reagent are invalid, need detect test again.
AC is at 0.7IU/ml or when above, and this reagent testing result is the critical positive or the positive; At 0.35IU/ml or when following, this reagent testing result is negative; In the time of between 0.35-0.7IU/ml, this reagent testing result is critical positive or negative.
What should explain at last is: the above is merely the preferred embodiment of the utility model; Be not limited to the utility model; Although the utility model has been carried out detailed explanation with reference to previous embodiment; For a person skilled in the art, it still can be made amendment to the technical scheme that previous embodiment is put down in writing, and perhaps part technical characterictic wherein is equal to replacement.All within the spirit and principle of the utility model, any modification of being done, be equal to replacement, improvement etc., all should be included within the protection domain of the utility model.
Claims (5)
1. a solid phase carrier that is used for enzyme linked immunosorbent assay (ELISA) is characterized in that, said solid phase carrier is can protein-bonded cellulose membrane.
2. the solid phase carrier that is used for enzyme linked immunosorbent assay (ELISA) according to claim 1 is characterized in that, said cellulose membrane is a nitrocellulose filter.
3. the solid phase carrier that is used for enzyme linked immunosorbent assay (ELISA) according to claim 2 is characterized in that, on the said nitrocellulose filter detection line can be set, said detection line position encapsulate can with the specificity anaphylactogen of IgE antibodies in the blood.
4. the solid phase carrier that is used for enzyme linked immunosorbent assay (ELISA) according to claim 3 is characterized in that, also can be provided with nature controlling line on the said nitrocellulose filter.
5. a kit that is used for enzyme linked immunosorbent assay (ELISA) is characterized in that, comprises each described solid phase carrier of claim 1-4.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011205430239U CN202433382U (en) | 2011-12-22 | 2011-12-22 | Solid phase carrier for enzyme-linked immunosorbent assay and kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011205430239U CN202433382U (en) | 2011-12-22 | 2011-12-22 | Solid phase carrier for enzyme-linked immunosorbent assay and kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN202433382U true CN202433382U (en) | 2012-09-12 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011205430239U Expired - Fee Related CN202433382U (en) | 2011-12-22 | 2011-12-22 | Solid phase carrier for enzyme-linked immunosorbent assay and kit |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111122861A (en) * | 2020-01-20 | 2020-05-08 | 浙江大学 | Kit for detecting specific IgE of serum mycoplasma pneumoniae |
-
2011
- 2011-12-22 CN CN2011205430239U patent/CN202433382U/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111122861A (en) * | 2020-01-20 | 2020-05-08 | 浙江大学 | Kit for detecting specific IgE of serum mycoplasma pneumoniae |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120912 Termination date: 20131222 |