CN202049160U - COxB (Coxsackievirus B) IgG/IgM (immunoglobulin G/immunoglobulin M) detection reagent kit by enzyme-linked immunosorbent assay method - Google Patents
COxB (Coxsackievirus B) IgG/IgM (immunoglobulin G/immunoglobulin M) detection reagent kit by enzyme-linked immunosorbent assay method Download PDFInfo
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- CN202049160U CN202049160U CN2011201373650U CN201120137365U CN202049160U CN 202049160 U CN202049160 U CN 202049160U CN 2011201373650 U CN2011201373650 U CN 2011201373650U CN 201120137365 U CN201120137365 U CN 201120137365U CN 202049160 U CN202049160 U CN 202049160U
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Abstract
The utility model discloses a COxB (Coxsackievirus B) IgG/IgM (immunoglobulin G/immunoglobulin M) detection reagent kit by an enzyme-linked immunosorbent assay method, which comprises a kit body. Eight recessed bottle slots, eight reagent bottles provided with sealing caps, a porous enzyme-linked plate and a plate sealing membrane are arranged in the kit body, the eight reagent bottles containing sample diluents, positive control liquid, negative control liquid, an IgG enzyme-labeled antibody, a substrate A, a substrate B, concentrate washing liquid and stopping solution are correspondingly arranged in the eight recessed bottle slots respectively, the porous enzyme-linked plate comprises a plurality of micro reaction holes with bottom surfaces sealed and top surfaces open, and COxB IgG/IgM antigen layers are coated on the inner walls of the micro reaction holes. The reagent kit can be used for detecting COxB IgG/IgM by the enzyme-linked immunosorbent assay method.
Description
Technical field
The utility model relates to CoxB IgG/IgM enzyme-linked immunosorbent assay (ELISA) kit.
Background technology
Coxsackie virus is a kind of of enterovirus, behind the Coxsackie virus infection, aseptic meningitis, flesh fiber crops low, herpangina, myocarditis and pericarditis, acute hemorrhagic conjunctivitis etc. can occur.This virus is divided into two groups of A, B again, and the B group has 6 serotypes.Whether the mensuration of CoxB IgG/IgM can detect the patient early and be infected recently, especially the myocarditis of acute stage is carried out the generaI investigation of CoxB IgG/IgM, to provide diagnosis basis to cardiomyopathies, the positive of CoxB IgM particularly, to point out the patient to be infected in early days by cardiac muscle, the favourable index that can cooperate simultaneously clinical diagnosis and observation has great significance to patient's treatment and rehabilitation.
The diagnostic method of Coxsackie virus infection mainly contains three kinds, viral isolation identification, Serological testing and Protocols in Molecular Biology.Virus separation and Culture and molecular biology method is complicated bothersome, and there is the risk of cross pollution in traditional RT-PCR technology length consuming time, can't satisfy the needs of handling great amount of samples during the viral prevalence simultaneously.Serological progress (express method) opened up should virus the quick diagnosis technology, it has the advantage of high sensitivity and high specific, brought into play more and more important effect in the diagnosis of virus infections.
The utility model content
The purpose of this utility model is to overcome defective of the prior art, and a kind of CoxB IgG/IgM enzyme-linked immunosorbent assay (ELISA) kit is provided.
The following technical proposals that adopted the utility model solves above-mentioned technical matters:
A kind of CoxB IgG/IgM enzyme-linked immunosorbent assay (ELISA) kit, comprise box body, box body is built-in with 8 recessed bottle, 8 bottles and is provided with the reagent bottle of gland bonnet, a porous elisa plate and a shrouding film, wherein, corresponding respectively each one bottle of the reagent bottle that sample diluting liquid, positive control, negative control, IgG enzyme labelled antibody, substrate A, substrate B, concentrated cleaning solution and stop buffer are housed of placing in described 8 recessed bottle positions, described porous elisa plate comprises several bottom surface sealing and uncovered little reacting holes of end face, and the inwall of little reacting hole is coated with CoxB IgG/IgM antigen layer.
CoxB IgG/IgM enzyme-linked immunosorbent assay (ELISA) kit is CoxB IgG or CoxB IgM enzyme-linked immunosorbent assay (ELISA) kit.
CoxB IgG/IgM antigen layer is CoxB IgG antigen layer or CoxB IgM antigen layer.
Described recessed bottle position can be made by polyfoam or cardboard.
Described porous elisa plate pad is under the reagent bottle or be positioned on the reagent bottle described shrouding film and described porous elisa plate placed adjacent.The size of described shrouding film is consistent with the end face size of porous elisa plate.
Further, described porous elisa plate comprises bracing frame (1), and is placed on several the removable micro reaction plate bars (2) on the bracing frame, and every micro reaction plate bar is provided with several removable little reacting holes (3).
Further, be provided with 48 or 96 little reacting holes on the described porous elisa plate altogether.
Further, described porous elisa plate is provided with big envelope outward.
Sample diluting liquid in the kit also can adopt conventional ELISA sample diluting liquid, as contains the sample diluting liquid of phosphate buffer, perhaps also can for contain the 0.01M phosphate buffer (PBS, pH7.2-7.5) and＜10% calf serum of 0.1%NaN3.
Positive control in the kit is the positive serum that contains CoxB IgG antibody or CoxB IgM antibody.
The negative serum of negative control in the kit.
IgG enzyme labelled antibody in the kit is the IgG antibody of horseradish peroxidase (HRP) mark, and IgG antibody can be various types of IgG antibody, as mouse-anti human IgG, goat anti-human igg or rabbit anti-human igg.
Substrate A in the kit, substrate B are conventional ELISA substrate A and substrate B.
Concentrated cleaning solution in the kit can adopt the conventional ELISA cleansing solution that concentrates, as contains the phosphate buffer of tween, perhaps also can be the concentrated 0.01MPBS of 20X.
Stop buffer in the kit can adopt conventional ELISA stop buffer.
The CoxB IgG/IgM antigen of bag quilt is purified CoxB IgG/IgM antigen (being CoxB IgG antigen or CoxB IgM antigen) on the porous elisa plate.
Wherein, the porous elisa plate that is coated with CoxB IgG/IgM antigen layer can adopt the mode of sealing with confining liquid behind the conventional antigen coated elisa plate to prepare.
Except that comprising above-mentioned substance, also can comprise needed reagent, instrument or container when conventional ELISA tests, in the kit as distilled water or deionized water, washing lotion drop bottle, test tube and instructions etc.
CoxB IgG/IgM enzyme-linked immunosorbent assay (ELISA) kit specificity of the present utility model and highly sensitive, the running time is short, can be used for many person-portions qualitative detection CoxB IgG/IgM.
Description of drawings
Fig. 1: the inner decomposition texture synoptic diagram of kit
Fig. 2: 48 hole elisa plate structural representations
Fig. 3: 96 hole elisa plate structural representations
Embodiment
Embodiment 1
CoxB IgG/IgM enzyme-linked immunosorbent assay (ELISA) kit, comprise box body, be equipped with 8 adjacent recessed bottle positions (11-18) in the box body as shown in Figure 1,8 bottles of reagent bottles that are provided with gland bonnet, a porous elisa plate 3 and a shrouding film 4, wherein, the corresponding respectively reagent bottle 21 that sample diluting liquid is housed of placing in described 8 recessed bottle positions, the reagent bottle 22 of positive control is housed, the reagent bottle 23 of negative control is housed, the reagent bottle 24 of IgG enzyme labelled antibody is housed, the reagent bottle 25 of substrate A is housed, the reagent bottle 26 of substrate B is housed, the reagent bottle 27 and respectively one bottle of the reagent bottle 28 that stop buffer is housed of concentrated cleaning solution are housed, described porous elisa plate 3 comprises several bottom surface sealing and uncovered little reacting holes of end face, and the inwall of little reacting hole is coated with CoxBIgG/IgM antigen layer.
Described porous elisa plate 3 fills up under each reagent bottle, described shrouding film 4 and described porous elisa plate 3 neighbouring placements, and the size of described shrouding film 4 is consistent with the end face size of porous elisa plate 3.The described porous elisa plate 3 outer big envelopes 5 that are provided with.
Described porous elisa plate 3 comprises bracing frame 31 as shown in Figures 2 and 3, and is placed on several the removable micro reaction plate bars 32 on the bracing frame, and every micro reaction plate bar is provided with several removable little reacting holes 33.
To 48 person-portion kits, be provided with 48 little reacting holes on the described porous elisa plate altogether, described sample diluting liquid is 5ml, positive control and negative control are 0.5ml, and the IgG enzyme labelled antibody is 5ml, and substrate A and substrate B are 2.5ml, concentrated cleaning solution is 20ml, and stop buffer is 2.5ml.
To the kit of 96 person-portions, be provided with 96 little reacting holes on the described porous elisa plate altogether, described sample diluting liquid is 10ml, positive control and negative control are 1ml, and the IgG enzyme labelled antibody is 10ml, and substrate A and substrate B are 5ml, concentrated cleaning solution is 30ml, and stop buffer is 5ml.
Porous elisa plate and all ingredients preparation in the kit:
1. sample diluting liquid: the 0.01M phosphate buffer (PBS, pH7.2-7.5),＜0.1%NaN
3, 10% calf serum.
2. positive control: contain＜0.1%NaN
3Positive human serum.The deactivation human serum is dissolved in protein-contg phosphate buffer; And having passed through hepatitis B virus surface antigen, human immunodeficiency virus (1,2 type) antibody, hepatitis C virus antibody, syphilis helicoid antibody are all negative.
3. negative control: contain＜0.1%NaN
3Negative human serum.The deactivation human serum is dissolved in protein-contg phosphate buffer; And having passed through hepatitis B virus surface antigen, human immunodeficiency virus (1,2 type) antibody, hepatitis C virus antibody, syphilis helicoid antibody are all negative.
4.IgG enzyme labelled antibody: be dissolved in PBS (pH7.2-7.5), contain (HRP) goat-anti people Ig and the carrier protein that contains antiseptic.
5. substrate A: contain the 0.1M sodium citrate, 0.01% hydrogen peroxide.
6. substrate B: contain tetramethyl benzidine, dimethyl sulfoxide, sodium acetate solution.
7. concentrated cleaning solution: (20X) concentrated 0.01MPBS.
8. stop buffer: 2mol/L H
2SO
4
9. be coated with the porous elisa plate of CoxB IgG/IgM antigen layer: adopt the mode of sealing with confining liquid behind the conventional antigen coated elisa plate to prepare.
The condition of storage of the utility model kit: 2～8 ℃ of preservations.
The using method of kit:
Kit can detect serum sample, collects serum by standard method, and is to be measured.As not measuring immediately, be placed on below-20 ℃ frozenly, and avoid multigelation.
1. reagent preparation
1.1 the preparation of solution: get 1 bottle of concentrated cleaning solution in the kit, with stand-by in the wash bottle (providing for oneself) of packing into after 1: 20 dilution mixing, the dissolving.
1.2 experiment is prepared: sample to be measured is taken out from refrigerator, put after room temperature thaws, sample to be measured is numbered by requirement of experiment.
1.3 after the careful confirmation in sample to be measured numbering back, with test serum with sample diluting liquid (diluting at 1: 100) promptly 10 μ l serum be added in the 1ml sample diluent mixing.
2. must satisfied experiment condition
2.1 kit is taken out in 2～8 ℃ of refrigerators, put room temperature and placed 30 minutes, the abundant mixing of reagent can be used.
2.2 the wavelength that uses microplate reader to measure is 450nm.
2.3 adjusting the temperature of water bath is 37 ℃.
3. the yin and yang attribute reference substance in the kit directly uses, and need not to dilute again, and every hole adds 100 μ l.
4. operation steps
Application of sample amount (μ l)
Add each 50 μ l/ hole of chromogenic substrate A liquid and B liquid, the mixing that gently shakes, the room temperature lucifuge was placed 10-15 minute, added stop buffer 50 μ l/ holes, slightly vibrated with mixed solution, placed room temperature 5 minutes.
In 15 minutes, detect, read each hole OD value in wavelength 450 nanometers with microplate reader.
5. quality control
The kit positive reference substance should be not less than 3 with the average relative OD ratio of negative control product.
6. the result calculates
Calculate the average OD value of each reference substance, sample, and judge the yin and yang attribute of sample according to the ratio of sample and negative control OD.
[kit Cut off value]
The average OD value of Cut off value=negative control X 2.1
Serum sample and ratio negative control OD value 〉=Cut off value positive (+);
Serum sample and ratio negative control OD value≤Cut off value positive (-).
Claims (7)
1. CoxB IgG/IgM enzyme-linked immunosorbent assay (ELISA) kit, comprise box body, it is characterized in that, box body is built-in with 8 recessed bottle positions, 8 bottles of reagent bottles that are provided with gland bonnet, a porous elisa plate and a shrouding film, corresponding respectively the placement is equipped with sample diluting liquid in described 8 recessed bottle positions, positive control, negative control, the IgG enzyme labelled antibody, substrate A, substrate B, each one bottle of the reagent bottle of concentrated cleaning solution and stop buffer, described porous elisa plate comprises several bottom surface sealing and uncovered little reacting holes of end face, and the inwall of little reacting hole is coated with CoxB IgG/IgM antigen layer.
2. CoxB IgG/IgM enzyme-linked immunosorbent assay (ELISA) kit according to claim 1 is characterized in that, described porous elisa plate pad is under the reagent bottle or place on the reagent bottle.
3. CoxB IgG/IgM enzyme-linked immunosorbent assay (ELISA) kit according to claim 1 is characterized in that described shrouding film and described porous elisa plate placed adjacent.
4. CoxB IgG/IgM enzyme-linked immunosorbent assay (ELISA) kit according to claim 1 is characterized in that, the size of described shrouding film is consistent with the end face size of porous elisa plate.
5. CoxB IgG/IgM enzyme-linked immunosorbent assay (ELISA) kit according to claim 1, it is characterized in that, described porous elisa plate comprises bracing frame, and is placed on several the removable micro reaction plate bars on the bracing frame, and every micro reaction plate bar is provided with several removable little reacting holes.
6. CoxB IgG/IgM enzyme-linked immunosorbent assay (ELISA) kit according to claim 1 is characterized in that, is provided with 48 or 96 little reacting holes on the described porous elisa plate altogether.
7. CoxB IgG/IgM enzyme-linked immunosorbent assay (ELISA) kit according to claim 1 is characterized in that described porous elisa plate is provided with big envelope outward.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011201373650U CN202049160U (en) | 2011-05-03 | 2011-05-03 | COxB (Coxsackievirus B) IgG/IgM (immunoglobulin G/immunoglobulin M) detection reagent kit by enzyme-linked immunosorbent assay method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011201373650U CN202049160U (en) | 2011-05-03 | 2011-05-03 | COxB (Coxsackievirus B) IgG/IgM (immunoglobulin G/immunoglobulin M) detection reagent kit by enzyme-linked immunosorbent assay method |
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| CN202049160U true CN202049160U (en) | 2011-11-23 |
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| CN2011201373650U Expired - Fee Related CN202049160U (en) | 2011-05-03 | 2011-05-03 | COxB (Coxsackievirus B) IgG/IgM (immunoglobulin G/immunoglobulin M) detection reagent kit by enzyme-linked immunosorbent assay method |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102590504A (en) * | 2012-01-31 | 2012-07-18 | 上海尧浩生物技术有限公司 | CoxB IgG/Ig/M test kit using enzyme linked immunosorbent assay |
| CN108519483A (en) * | 2018-05-26 | 2018-09-11 | 江苏力博医药生物技术股份有限公司 | Based on the platelet antibody detection kit and its detection method of solid agglutination method and application |
| CN111999495A (en) * | 2020-08-18 | 2020-11-27 | 北京贝尔生物工程股份有限公司 | Kit for detecting Coxsackie group B virus IgM antibody by magnetic particle chemiluminescence method and preparation method thereof |
| CN112630453A (en) * | 2020-12-23 | 2021-04-09 | 贵州金域医学检验中心有限公司 | Constant temperature kit for gynecological tumor marker detection and detection method thereof |
| CN115032151A (en) * | 2022-06-10 | 2022-09-09 | 杨富松 | Efficient multifunctional enzyme-labeling instrument and use method thereof |
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2011
- 2011-05-03 CN CN2011201373650U patent/CN202049160U/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102590504A (en) * | 2012-01-31 | 2012-07-18 | 上海尧浩生物技术有限公司 | CoxB IgG/Ig/M test kit using enzyme linked immunosorbent assay |
| CN102590504B (en) * | 2012-01-31 | 2015-01-14 | 上海尧浩生物技术有限公司 | CoxB IgG/Ig/M test kit using enzyme linked immunosorbent assay |
| CN108519483A (en) * | 2018-05-26 | 2018-09-11 | 江苏力博医药生物技术股份有限公司 | Based on the platelet antibody detection kit and its detection method of solid agglutination method and application |
| CN111999495A (en) * | 2020-08-18 | 2020-11-27 | 北京贝尔生物工程股份有限公司 | Kit for detecting Coxsackie group B virus IgM antibody by magnetic particle chemiluminescence method and preparation method thereof |
| CN112630453A (en) * | 2020-12-23 | 2021-04-09 | 贵州金域医学检验中心有限公司 | Constant temperature kit for gynecological tumor marker detection and detection method thereof |
| CN115032151A (en) * | 2022-06-10 | 2022-09-09 | 杨富松 | Efficient multifunctional enzyme-labeling instrument and use method thereof |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20111123 Termination date: 20200503 |