CN202153226U - Joint-detecting test paper for rubella virus IgM and IgG antibody - Google Patents
Joint-detecting test paper for rubella virus IgM and IgG antibody Download PDFInfo
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- CN202153226U CN202153226U CN2011201861033U CN201120186103U CN202153226U CN 202153226 U CN202153226 U CN 202153226U CN 2011201861033 U CN2011201861033 U CN 2011201861033U CN 201120186103 U CN201120186103 U CN 201120186103U CN 202153226 U CN202153226 U CN 202153226U
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Abstract
The utility model discloses a piece of joint-detecting test paper for a rubella virus IgM and an IgG antibody, which is characterized in that one end of a plastic support plate (8) is pasted with an upper sample pad (1), one end of the upper sample pad (1) is tightly connected with a collaurum pad (2) containing a antigen gpE1 signed with rubella virus specificity in a pressing way, one end of the collaurum pad (2) is tightly connected with a nitrocellulose NC film (3) in the pressing way, a detecting line T1 (5), a detecting line T2 (6) and a quality control C line which are mutually separate are coated on the film, the detecting line T1 (5) is an anti-human IgM antibody, the detecting line T2 (6) is another anti-human IgG antibody, the quality control C line (7) is an anti-rubella virus gpE1 antibody, and the other end of the NC film is connected with a piece of sample absorbing pad forming test paper (4). A sample is added on the upper sample pad during detection, and is directly observed by naked eyes within 15mins, and detection results are judged. The joint-detecting test paper for the rubella virus IgM and the IgG antibody has the advantages of being simple and fast in operation, suitable for site diction, economical and practical, and the like.
Description
Technical field
The utility model relates to the biologic applications technical field, particularly relates to a kind of with colloidal gold immunity chromatography fast detecting rubella virus IgM and the antibody combined detection test paper of IgG
Background technology
Rubella is a kind ofly to pass through airborne acute infectious disease by what rubella virus caused, can get in the fetus body through placenta, cause fetal anomaly.And the time that the pregnant woman infects rubella virus more early, and the possibility of teratogenesis is big more.Serological research shows, is infected even infect the women of child-bearing age of rubella virus again, also can not work the mischief to fetus after the pregnancy.But have 10% the women of child-bearing age to belong to the rubella susceptible person approximately, if the primary infection rubella virus, the fetus terateger rate is up to more than 80% in 3 months of pregnancy for they.Many in the world developed countries classify the inspection that rubella infects as pregnancy period routine inspection project, aspect prenatal and postnatal care, are bringing into play important effect.
The main method that at present rubella virus is detected is that the rubella virus specific antibody detects.Detect rubella virus IgM and take a decision as to whether the infection of existing disease, detect IgG and judge whether once infected.Detect two kinds of antibody and help the carrying out of checked object comprehensively judged, instruct prenatal and postnatal care.Rubella virus detection of antibodies method mainly is detection methods such as ELISA at present.In the detection method of collaurum, the report of independent detection rubella virus IgM or IgG is arranged, but detect rubella virus IgM simultaneously and IgG detection of antibodies test paper does not have relevant report as yet with collaurum para-immunity chromatographic technique.With detect IgM respectively with two kinds of reagent and compare with IgG antibody, detect the workload that IgM and IgG antibody capable reduce the testing staff simultaneously, practice thrift cost; Reduce blood sampling volume simultaneously, reduce patient's misery.
The immunochromatography colloidal gold technique is novel diagnostic techniques; Aspect antibody test, obtained comparatively extensively using, ultimate principle is following: utilize a kind of antigen of colloid gold label or antibody, on the NC of reagent film, encapsulate corresponding pairing antigen or antibody; During detection when containing corresponding specific antibody or antigen in the sample; The part formation compound that combines in colloid gold label particle and the sample, chromatography on the NC film then, coated again antigen or antibody capture; Form macroscopic detection T line, have or not the judgement of realization the result through detection line.Have easy and simple to handle, reaction fast, high, the high specificity of susceptibility, be fit to on-the-spot the detection and advantage such as economical and practical.
Summary of the invention
The purpose of the utility model provides a kind of rubella virus IgM and IgG antibody test test paper, have easy and simple to handle, reaction fast, high, the high specificity of susceptibility, be fit to on-the-spot the detection and advantage such as economical and practical.
The utility model provides a kind of rubella virus IgM and the antibody combined detection test paper of IgG; It is characterized in that plastic support board (8) one ends connect upward appearance pad (1); The tight crimping of one end of last appearance pad (1) contains the collaurum pad (2) of the specific antigen gpE1 of underlined rubella virus; Collaurum pad (2) one ends tight crimping cellulose nitrate NC films (3); Nitrocellulose filter is coated with detection line T1 (5), detection line T2 (6) and the Quality Control C line that is separated from each other, and detection line T1 (5) is for being coated on the anti-human IgM antibody on the NC film, and detection line T2 (6) is for being coated on the anti-human IgG antibody on the NC film; Quality Control C line (7) is for being coated on the wind resistance exanthema virus gpE1 antibody on the NC film, and the other end of nitrocellulose filter connects suction appearance pad (4) and forms test paper.
The antibody combined detection test paper of described rubella virus IgM and IgG is characterized in that the specific antigen gpE1 of described antigen rubella virus antigen is the antigen that utilizes genetic engineering recombinant expressed.
The antibody combined detection test paper of described rubella virus IgM and IgG is characterized in that the described appearance pad (1) of going up is glass fibre membrane or nonwoven fabrics, inhales appearance pad (4) and is made up of absorbent filter.
The antibody combined detection test paper of described rubella virus IgM and IgG; The method for coating of described antigen is: will resist human IgM antibody to be mixed with the solution of 1-2mg/ml with 0.01M pH 7.2 phosphate buffers (PBS), will resist the human IgG antibody to be mixed with the solution of 1-2mg/ml with 0.01M pH 7.2 phosphate buffers (PBS), and rule with the parameter of 1-1.5ul/cm in NC film bottom with spray film appearance; Encapsulate T1, T2 line; Simultaneously encapsulate wind resistance exanthema virus gpE1 antibody as the C line on NC film top, after the line with the NC film at drying room, temperature 20-25 ℃; Humidity is less than 30%, dry 2-5 hour.
The antibody combined detection test paper of described rubella virus IgM and IgG; The method of described antigenic mark colloid gold particle is: prepare the colloidal gold solution that diameter is 30-50nm with gold chloride-trisodium citrate reduction method; Get 100ml collaurum liquid after preparation is accomplished and be placed in the beaker, use 0.2M K
2CO
3Transfer to pH8.5, press the 100ml colloidal gold solution and add 0.5-2mg rubella virus gpE1 antigen, stirring at room 2 hours; Add 1% bovine serum albumin(BSA) BSA; 1% polyglycol PEG20000 seals 20min, and centrifugal 30 minutes of 12000r/m abandons supernatant; Redissolve to 100ml with the collaurum working fluid, press 1ml solution shop 20cm
2Ratio be layered on equably on glass fibre membrane or the nonwoven fabrics, put drying room again, temperature 20-25 ℃, humidity dry 2-5 hour, is processed the collaurum pad less than 30%.
The antibody combined detection test paper of described rubella virus IgM and IgG, the assembly method of described test paper is: in hothouse, temperature 20-25 ℃; Humidity is less than 40%; Get the plastic support board plate, paste at the middle part that the NC film that has encapsulated is placed on plastic support board, pastes at NC film T line one side overlap joint collaurum pad (take collaurum pad 1/3); Paste the appearance pad at collaurum pad opposite side overlap joint, take 1/5 of collaurum pad; Inhale the appearance pad at NC film C line one side overlap joint, take and inhale 1/10 of appearance pad; To post plastic plate with cutter at last and be cut into the wide test strips of 2-5mm, the test strips that cuts can reinstall in the plastic clip, forms test card.
The antibody combined detection test paper of described rubella virus IgM and IgG; Detection method is: seized serum or blood plasma balance to the greenhouse, are kept flat test strips or reagent card, on last appearance pad, add the 5-20ul test sample; The sample diluting liquid that adds 50-100ul again; Make collaurum dissolving and on the NC film chromatography, the appearance situation of Direct observation C, T1, T2 line in 15 minutes with the naked eye then, and judge testing result.
The beneficial effect of the utility model is: a kind of novel test paper that utilizes the immunochromatography colloidal gold technique to detect rubella virus IgM and IgG antibody is provided.Adopt colloid gold label rubella virus specific antigen gpE1, encapsulate anti-people IgM and IgG antibody and realize that to wind resistance exanthema virus detection of antibodies in the blood one-time detection obtains two kinds and detects index, saves time and cost, and alleviates patient's misery.Have easy and simple to handle, reaction fast, high, the high specificity of susceptibility, be fit to on-the-spot the detection and advantage such as economical and practical.
Description of drawings:
The antibody combined detection test paper of Fig. 1 rubella virus IgM and IgG structural representation.
The reference numeral explanation:
1: go up the appearance pad; 2: the collaurum pad; The 3:NC film; 4: inhale the appearance pad
5: detection line T1; 6: detection line T2; 7: Quality Control C line; 8: plastic support board
Embodiment
Embodiment: preparation rubella virus IgM and the antibody combined detection test paper of IgG
1 main material
1.1 rubella virus specificity gpE1 antigen, gpE1 antibody: Central Plains, Beijing Company products, be recombinant antigen, gpE1 antigen is used for colloid gold label, and gpE1 antibody is used for NC film nature controlling line and encapsulates; Mouse-anti people IgM and IgG antibody: the Arista Company products is used for the NC film and encapsulates; Gold chloride: Sigma Company products; Cellulose nitrate (NC) film: Millipore Company products; Bovine serum albumin(BSA) (BSA), polyglycol PEG20000, caseinhydrolysate: Sigma product.Other common agents is AR.
1.2 clinical serum is obtained in relevant hospital by company, wherein the rubella virus IgG antibody positive sample is 80 parts, and 20 parts in IgM antibody positive sample is comprising 15 parts of two kinds of equal positive sample of antibody.50 parts in two kinds of complete negative normal person's samples of antibody.
1.3 rubella virus IgM antibody detection kit (ELISA method), rubella virus IgG antibody detection kit (ELISA method): domestic registered commercial product.
2 methods
2.1 the colloid gold label gold chloride-trisodium citrate reduction method of rubella virus gpE1 recombinant antigen prepares the colloidal gold solution that diameter is 40nm, preparation is got three parts of collaurums after accomplishing, and uses 0.2M K respectively
2CO
3Solution is transferred to pH7.5, pH8.5 and pH9.0.Then solution is placed on the magnetic stirring apparatus and slowly stir; Add 0.5mg, 1mg, 1.5mg by every 100ml solution recombinant antigen slowly is added drop-wise to albumen in the colloidal gold solution, continue to stir 2 hours, be added dropwise to final concentration again and be 0.1% PEG2000 and 1% BSA and seal 20min; It is centrifugal with 12000r/m that mark finishes the back; Abandon supernatant, deposition is pressed original volume and is redissolved to the collaurum working fluid borate buffer solution of different proportionings, pH8.0; Contain BSA, sheep blood serum, in sucrose and the surfactant.Then the mark colloidal gold solution is pressed 1ml solution shop 20cm
2The ratio application of sample on nonwoven fabrics, at temperature 20-25 ℃, relative humidity is processed the collaurum pad dry 2-4 hour of＜30% drying room.
2.2NC film encapsulates with 0.01M pH7.2PBS mouse-anti people IgM is diluted to 0.5mg/ml, 1mg/ml, 2mg/ml respectively; The mouse-anti human IgG is diluted to 0.5mg/ml, 1mg/ml, 2mg/ml respectively; Ruling respectively by 1ul/cm in NC film bottom with spray film appearance then encapsulates, and encapsulates anti-gpE1 antibody on NC film top simultaneously, is used for the Quality Control of product; Encapsulate after the completion the NC film at temperature 20-25 ℃, relative humidity was dry 2-5 hour of＜30% drying room.
2.3 the antibody combined detection test paper of rubella virus IgM and IgG be assembled in the hothouse (temperature 20-25 ℃; Relative humidity＜30%) gets plastic support board; Paste at the middle part that the NC film that has encapsulated is placed on plastic support board; Paste at NC film T line one side overlap joint collaurum pad (take collaurum pad 1/3), paste appearance pad (take collaurum pad 1/5) at collaurum pad opposite side overlap joint; Inhale appearance pad (take inhale appearance pad 1/10) at NC film C line one side overlap joint; Plastic plate be will post with cutter then and 3mm or the wide test strips of 4mm will be cut into.The test strips that cuts can reinstall in the plastic clip, forms rubella virus IgM and the antibody combined detectable card of IgG.
2.4 detection method to the greenhouse, keeps flat seized serum or blood plasma balance with test strips for preparing or reagent card, on last appearance pad, add the 10ul test sample; If contain wind resistance exanthema virus IgM or IgG antibody in the sample, then with sample pad on the collaurum of mark gpE1 antigen combine, form compound; And be diffused on the NC film further chromatography; When running into the mouse-anti people IgM that is coated on T1 line place on the NC film, compound then again with encapsulate anti-IgM antibodies, be trapped in and encapsulate the place; When running into when being coated on the NC film T2 line IgG of place, compound then combines with coated antibody again, is trapped in and encapsulates line T2 place; When captive colloidal gold composite reaches some, then form a macroscopic T1 or T2 line; If do not contain specific antibody in the serum, then can not form immune complex, also can not form the T line, the C line is as the quality control standard of reagent, and positive and negative sample all can occur when detecting.The appearance situation of Direct observation C, T line in 5,10,15,20 and 30 minutes with the naked eye, and judgement testing result.
2.5 test paper technological parameter debugging with concentration not isolabeling, the reagent that encapsulates make up pairing, the preparation sample utilizes quality controlled serum that reagent is tested, and finds best of breed.
2.6 clinical serum detects and all clinical serum is detected by detection method with this reagent, carries out control test with the commercial ELISA reagent that detects simultaneously.
3 results
3.1 the test paper parameter is confirmed the testing result according to sample, the optimum mark pH value of having confirmed test paper is 8.0; The optimum mark amount of reorganization hybrid antigen is the 1mg/100ml colloidal gold solution; Best collaurum working fluid is a 10mM borosilicate damping fluid, and pH8.5 contains 0.5%BSA, 10% sheep blood serum, 2% sucrose, and 0.2%Tween 20; Best bag mouse-anti people IgM and IgG encapsulate concentration and are 1.5mg/ml.The optimal decision time of testing result is 5-15 minute.But above parameter possibly need suitably adjustment when preparation different batches product.
3.2 it is contrast that clinical serum detects with ELISA reagent, and IgG antibody is detected, this reagent IgG detects positive 78 parts, and ELISA detects positive 80 parts of IgG, sensitivity=78/80=97.5%; This reagent IgG negatives detects all negative, relative specificity 100%.P＞0.05。With ELISA reagent is contrast, and IgM antibody is detected, and this reagent IgM detects positive 19 parts, and ELISA detects positive 20 parts of IgM, sensitivity=19/20=95.0%; This reagent IgM negatives detects all negative, relative specificity 100%.P＞0.05。Simultaneously positive this reagent of sample of IgM and IgG detect and ELISA reagent in full accord.Clinical detection shows that the performance of this reagent compares there was no significant difference with ELISA reagent, is suitable for Clinical detection.
Claims (3)
1. rubella virus IgM and the antibody combined detection test paper of IgG; It is characterized in that plastic support board (8) one ends connect upward appearance pad (1); The tight crimping of one end of last appearance pad (1) contains the collaurum pad (2) of the specific antigen gpE1 of underlined rubella virus; Collaurum pad (2) one ends tight crimping cellulose nitrate NC films (3); Nitrocellulose filter is coated with detection line T1 (5), detection line T2 (6) and the Quality Control C line that is separated from each other, and detection line T1 (5) is for being coated on the anti-human IgM antibody on the NC film, and detection line T2 (6) is for being coated on the anti-human IgG antibody on the NC film; Quality Control C line (7) is for being coated on the wind resistance exanthema virus gpE1 antibody on the NC film, and the other end of NC connects suction appearance pad (4) and forms test paper.
2. the antibody combined detection test paper of rubella IgM according to claim 1 and IgG is characterized in that described antigen rubella virus specific antigen gpE1 is the antigen that utilizes genetic engineering recombinant expressed.
3. the antibody combined detection test paper of rubella virus IgM according to claim 1 and IgG is characterized in that the described appearance pad (1) of going up is glass fibre membrane or nonwoven fabrics, inhales appearance pad (4) and is made up of absorbent filter.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011201861033U CN202153226U (en) | 2011-06-03 | 2011-06-03 | Joint-detecting test paper for rubella virus IgM and IgG antibody |
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| Application Number | Priority Date | Filing Date | Title |
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| CN2011201861033U CN202153226U (en) | 2011-06-03 | 2011-06-03 | Joint-detecting test paper for rubella virus IgM and IgG antibody |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102928587A (en) * | 2012-11-16 | 2013-02-13 | 南京凯基生物科技发展有限公司 | Colloidal gold method detection test strip and reagent kit for IgM and IgG antibodies of mycoplasma pneumoniae and preparation method of reagent kit |
| CN104330569A (en) * | 2014-10-29 | 2015-02-04 | 武汉生命科技股份有限公司 | Test strip for quickly detecting antibody |
-
2011
- 2011-06-03 CN CN2011201861033U patent/CN202153226U/en not_active Expired - Lifetime
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102928587A (en) * | 2012-11-16 | 2013-02-13 | 南京凯基生物科技发展有限公司 | Colloidal gold method detection test strip and reagent kit for IgM and IgG antibodies of mycoplasma pneumoniae and preparation method of reagent kit |
| CN104330569A (en) * | 2014-10-29 | 2015-02-04 | 武汉生命科技股份有限公司 | Test strip for quickly detecting antibody |
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| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CX01 | Expiry of patent term |
Granted publication date: 20120229 |
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| CX01 | Expiry of patent term |