CN204203235U - Nettle rash vaccine effect of inoculation Fast Evaluation kit - Google Patents
Nettle rash vaccine effect of inoculation Fast Evaluation kit Download PDFInfo
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- CN204203235U CN204203235U CN201420636037.9U CN201420636037U CN204203235U CN 204203235 U CN204203235 U CN 204203235U CN 201420636037 U CN201420636037 U CN 201420636037U CN 204203235 U CN204203235 U CN 204203235U
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Abstract
The utility model belongs to field of biological detection, is specifically related to a kind of nettle rash vaccine effect of inoculation Fast Evaluation kit.Described kit comprises box body, the container of sample diluting liquid and Test paper is housed; described Test paper is made up of base plate (1) and mutual closely overlap joint sample pad (6), antigen pad (5), label pad (4), coated film (3) and the absorption pad (2) be pasted onto on base plate; according to the rubella antibody minimal protection titre levels that WHO recommends; the lowest detection line of kit is set as 10IU/ml, for detecting the rubella virus IgG antibody in serum/plasma/whole blood.Kit described in the utility model is with low cost, and preparation technology is simple, and the detection result of serum antibody and the result of ELISA kit are fitted without significant difference, can be used for the Fast Evaluation of the rear immune effect of nettle rash vaccine inoculation.
Description
Technical field
The utility model belongs to field of biological detection, is specifically related to a kind of nettle rash vaccine effect of inoculation Fast Evaluation kit.
Background technology
Rubella is a kind of common acute infectious disease caused by rubella virus, with low-heat, skin rash for feature, often with after ear, occipital lymph node enlargement.If first trimester pregnancy infects rubella virus, can cause congenital rubella syndrome (CRS), 90% there will be multiple fetal anomaly, and usually causes miscarriage and stillbirth, and risk declines afterwards.Infection after 16 pregnant weeks seldom can cause fetal anomaly, still can cause fetus sensorineural hearing loss although infect within 20 pregnant weeks.Rubella is global distribution, and have seasonality (as occurred frequently in Temperate Region in China winter-spring season), every 5-9 is once popular.Rubella virus is a kind of enveloped virus, belongs to rubella virus section, is single strand RNA virus, only has One serotype, with other enveloped virus no cross reaction.The mankind are known unique hosts.Rubella virus is by respiratory infectious, and copy at mucous membrane of nasopharynx and regional nodes, latent period is 12-23 days, average 18 days.After contact virus, within 5-7 days, there will be viremia virusemia, viral spread is to Different Organs.Virus also can diaplacental infection fetus.
The effective measures that prevention rubella is popular mainly inoculate nettle rash vaccine.The expansion National immunization Program of China lists nettle rash vaccine in routine immunization program, the nettle rash vaccine gone on the market at present has the Measles-Parotitis Rubella combined vaccine (MMR) of unit price, the divalence of combining with measles or mumps vaccine or trivalent, and annual nearly tens million of children accept immunity inoculation.This points out us, is necessary to evaluate the herd immunity effect of nettle rash vaccine.Evaluate the content that the most reliable method of immune effect of vaccine is exactly the specific antibody detected in inoculator's body in serum.Vehicles Collected from Market detects commercial kit many employings euzymelinked immunosorbent assay (ELISA) and the chemoluminescence method of rubella antibody, although these methods are highly sensitive, result is accurate, chemical illuminating reagent can also carry out quantitatively by antagonist, but in practical operation, there is inconvenience, as step is complicated, consuming time longer, also need to use specific instrument and equipment; Kit needs 4 DEG C of preservations; To sample requirement venous blood collection etc., not easily in the universal utilization of basic unit.
Immune colloid gold quick diagnosis technology is based upon in enzyme linked immunosorbent assay, latex agglutination test, monoclonal antibody technique and immuno-gold labeling technical foundation, take collaurum as label, utilize special antigen-antibody reaction iodine signal, just can the new technology of result of determination by directly observing.This technology has the advantages such as simple, quick, accurate and pollution-free, many diagnostic fields such as to detect fast develop rapidly at clinical medicine detection, animal medicine detection, hormone test, food safety detection, medicament residue and drugs.At present, the Test paper of the quick detection antibody of colloid gold label is all made up of sample pad, label pad, coated film (it is drawn and has detection line, nature controlling line) and absorption pad, difference according to Cleaning Principle is divided into two kinds substantially: one is by antigen standard gold, label pad wraps by the antigen of colloid gold label, coated film draws film with the antibody of test antibodies and obtains detecting T line; Another kind is that antigen is drawn film, label pad wraps by the antibody of the test antibodies of colloid gold label, coated film draws film with antigen and obtains detecting T line.Now there are some researches show: because antibody protein character is comparatively stable, so the colloid gold label of antagonist albumen is comparatively stable, there is a lot of problem in the colloid gold label of the antigen therefore described in above-mentioned first method, because antigen mostly is virus type, character is various, some sizes are even also large than colloid gold particle, and less stable, be therefore difficult to directly by antigenic mark on colloid gold particle; Even if by antigenic mark, can also need the purity that antigen reaches high reluctantly, and to resist proviral purifying be the engineering that a difficulty is comparatively large, cost is higher; Also someone finds the recombinant antigen of antigen to replace these native antigens, but a lot of antigen is difficult to seek the recombinant antigen that can substitute, even and if have, cost is also high, illustrates thus, and the limitation of above-mentioned first method is higher.Above-mentioned second method uses the antibody of test antibodies to carry out gold mark, the problem of antigen gold mark can be avoided, but on coated film, use antigen to draw film obtain detecting T line, need antigen to have higher concentration and purity, which increases cost and the difficulty of production; In addition because Antigen Stability is poor, antigen is difficult to long-time preservation after drawing film, and therefore the term of validity of test strips is extremely short.These are all the reasons of restriction antibody quick detection type kit fast development.
Summary of the invention
The utility model is for the deficiencies in the prior art, object is to provide a kind of nettle rash vaccine effect of inoculation Fast Evaluation kit, carry out colloidal gold method with this kit to the rubella virus IgG antibody in serum in inoculator's body to detect fast, the Fast Evaluation to nettle rash vaccine effect of inoculation can be realized.
The technical scheme that the utility model adopts is:
Nettle rash vaccine effect of inoculation Fast Evaluation kit, comprise box body, container and the Test paper of sample diluting liquid are housed, described sample diluting liquid is housed container and Test paper be positioned at box body, it is characterized in that, described Test paper is by base plate and mutually closely overlaps the sample pad be pasted onto on base plate, antigen pad, label pad, coated film and absorption pad composition, described antigen pad is coated with rubella antigens, described label pad is coated with the mouse-anti rubella monoclonal antibody of colloid gold label, described coated film is fixed with and detects T line and Quality Control C line, described detection T line is coated with mouse-anti human IgG antibody, described Quality Control C line is coated with sheep anti-mouse igg antibody, the lowest detection line of described Test paper is 10IU/ml.
By such scheme, described sample diluting liquid is the mixed aqueous solution of phosphoric acid disodium hydrogen, sodium dihydrogen phosphate, sodium chloride and Sodium azide.
By such scheme, described antigen pad is glass fibre, nonwoven fabrics or polyester film.
By such scheme, described base plate is PVC base plate, and the one side of described PVC base plate is double faced adhesive tape.
By such scheme, described coated film is nitrocellulose filter.
By such scheme, described label pad and sample pad are glass fibre, and described absorption pad is thieving paper.
By such scheme, the preparation method of described antigen pad is: use antigen protection liquid to be carried out by rubella antigens diluting rear uniform spreading on glass fibre, nonwoven fabrics or polyester film, be placed in 37 DEG C, relative humidity is lower than in the environment of 40%, dry 4 hours, make antigen pad; The formula of described antigen protection liquid is: Na
2hPO
45.72g, NaH
2pO
40.624g, trisodium citrate 8.5g, sucrose 10g, Sodium azide 1g, water is settled to 1L; Described rubella antigens is crude antigen, and wherein the content of antigen is 40wt% ~ 50wt%.
In the utility model, in described kit, the Cleaning Principle of Test paper is: add serum or blood plasma or whole blood sample in sample pad, add 2 ~ 3 sample diluting liquids again, if containing rubella virus IgG antibody in sample, it forms rubella antigens-rubella virus IgG antibody compound by with the rubella antigens in antigen pad, this compound moves forward, the mouse-anti rubella monoclonal antibody of the colloid gold label of the rubella antigens in compound again in label pad is combined, form the mouse-anti rubella monoclonal antibody immunity compound of rubella virus IgG antibody-rubella antigens-colloid gold label, this compound moves forward along paper slip due to chromatography effect, to detecting T line, rubella virus IgG antibody in compound with detect mouse-anti human IgG antibody T line wrapping quilt and react the mouse-anti rubella monoclonal antibody immunity compound forming mouse-anti human IgG antibody-rubella virus IgG antibody-rubella antigens-colloid gold label, this compound can be gathered in detection zone, when the compound assembled reaches certain quantity, then form a macroscopic band (T line), judged result is positive, if do not contain in sample or containing the rubella virus IgG antibody seldom measured, then can not form immune complex, namely can not form macroscopic band, judged result is negative, C line is as the quality control standard of reagent, and positive and negative detection sample all can produce band.
Simultaneously; in the serum recommended according to the World Health Organization (WHO) (WHO), the minimal protection antibody titer levels of rubella virus is 10IU/ml; namely as the concentration >=10IU/ml of Detecting Rubella Virus Antibodies In Human Sera in inoculator's body; illustrate that nettle rash vaccine is inoculated successfully; as the concentration < 10IU/ml of Detecting Rubella Virus Antibodies In Human Sera in inoculator's body, illustrate that nettle rash vaccine inoculation is unsuccessful.In the utility model, the sensitivity of kit is set to 10IU/ml, the testing result of rubella virus IgG antibody is that namely the positive shows Rubella vaccine immunization success, and the testing result of rubella virus IgG antibody is that namely feminine gender shows that Rubella vaccine immunization is unsuccessful.Therefore, kit described in the utility model can realize the effect of inoculation of Fast Evaluation nettle rash vaccine.
The using method of the utility model kit: Test paper is kept flat, 5 μ l blood samples are added in sample pad, add 2 ~ 3 sample diluting liquids again, with the naked eye directly observe the colour developing situation detecting T line and Quality Control C line in 20 minutes: Quality Control C line does not develop the color, and this test failure is described; Quality Control C line develops the color, and detection T line does not develop the color and illustrates that nettle rash vaccine inoculation is unsuccessful; Quality Control C line and detection T line all develop the color and illustrate that nettle rash vaccine is inoculated successfully.
The beneficial effects of the utility model:
(1) compared with prior art, Test paper described in the utility model adds the structure of antigen pad, by adding antigen protection liquid and wrap by rubella virus antigen on antigen pad, label pad wraps by mouse-anti rubella monoclonal antibody, solve the problem that rubella virus antigen is not easy due to poor stability to be marked by gold; On the other hand, the utility model adopts mouse-anti human IgG antibody to draw film and obtains detecting T line, because antibody is compared comparatively stable, therefore the term of validity of test strips is longer.
(2) rubella virus antigen described in the utility model is crude antigen, do not need to be further purified crude antigen just can reach reaction effect, this is because in test strips reaction chromatography process, by double antibody sandwich method, screening and purifying are carried out to crude antigen, ensure that the specificity of reagent reacting, therefore the preparation cost of Test paper is low, is beneficial to popularization; Experimental result shows, utilize Test paper described in the utility model to detect antibody, the result of detection is consistent with ELISA testing result.
(3) compared with prior art, kit described in the utility model has quick, easy, sensitive, special feature, break away from the particular/special requirement of reagent to equipment, instrument, personnel and Storage Box transportation environment, the sensitivity of kit is set to 10IU/ml, the rapid evaluation to nettle rash vaccine effect of inoculation can be realized.
Accompanying drawing explanation
Fig. 1 is kit one-piece construction schematic diagram of the present utility model, and wherein A-box body, B-is equipped with the container of sample diluting liquid, C-Test paper, D-instructions.
Fig. 2 is the structural representation of Test paper in kit of the present utility model, and wherein, 1-base plate, 2-absorption pad, 3-coated film, 4-label pad, 5-antigen pad, 6-sample pad, 7-Quality Control C line, 8-detects T line.
Embodiment
In order to understand the utility model better, illustrate content of the present utility model further below in conjunction with embodiment, but content of the present utility model is not only confined to example below.
As shown in Figure 1, nettle rash vaccine effect of inoculation Fast Evaluation kit, comprise box body A, container B and the Test paper C of sample diluting liquid are housed, described sample diluting liquid is housed container B and Test paper C be positioned at box body A, described Test paper is by base plate 1 and mutually overlaps the sample pad 6 be pasted onto on base plate, antigen pad 5, label pad 4, coated film 3 and absorption pad 2 form, described antigen pad is coated with rubella antigens, described label pad is coated with the mouse-anti rubella monoclonal antibody of colloid gold label, described coated film is fixed with and detects T line 8 and Quality Control C line 7, described detection T line is coated with mouse-anti human IgG antibody, described Quality Control C line is coated with sheep anti-mouse igg antibody, the lowest detection line of described Test paper is 10IU/ml.
Instructions D is placed with in box body A.
Described sample diluting liquid is the mixed aqueous solution of phosphoric acid disodium hydrogen, sodium dihydrogen phosphate, sodium chloride and Sodium azide.In following examples, the component of Sample dilution is sodium hydrogen phosphate 5.72g, and sodium dihydrogen phosphate 0.624g, sodium chloride 8g, Sodium azide 0.5g, ultrapure water is settled to 1L.
Described base plate is PVC base plate, and the one side of described PVC base plate is double faced adhesive tape.
Described coated film is nitrocellulose filter; Described antigen pad is glass fibre, nonwoven fabrics or polyester film.
Described label pad and sample pad are glass fibre, and described absorption pad is thieving paper.
Described antigen pad obtains on glass fibre, nonwoven fabrics or polyester film by using antigen protection liquid that rubella antigens is carried out diluting rear uniform spreading, and described antigen is crude antigen, and wherein the content of antigen is 40wt% ~ 50wt%.
Embodiment 1 nettle rash vaccine effect of inoculation Fast Evaluation kit
Main material
Antigen: rubella virus antigen, purchased from microbix company, the natural viral of cultivation, for slightly pure, but deactivation, not there is biohazard.For the preparation of antigen pad.
Mouse-anti human IgG antibody, purchased from Xiamen Bo Sheng Bioisystech Co., Ltd, for the bag quilt of nitrocellulose filter T line.
Mouse-anti rubella monoclonal antibody, purchased from Wuhan Saixin Biological Technology Co., Ltd., for marking collaurum.
Sheep anti-mouse igg antibody, purchased from Hangzhou Long Ji Bioisystech Co., Ltd, for the bag quilt of nitrocellulose filter Quality Control C line.
Gold chloride: sigma Products.
Nitrocellulose filter: millipore Products.
Bovine serum albumin(BSA) (BSA), casein: sigma Products.
All the other chemical reagent are analytical reagent.
Clinical serum is obtained at Wuhan City's Blood Center by company, wherein positive 192 parts of rubella virus IgG antibody, IgG antibody negative sample 108 parts.
Rubella virus IgG antibody detection kit (ELISA method): purchased from Beijing Beier Bioengineering Co., Ltd..Domestic registered commercial product.
2, the preparation of label pad
Gold chloride-trisodium citrate reduction method prepares the colloidal gold solution that diameter is 40nm, after having prepared, get 100ml colloidal gold solution to be placed on magnetic stirring apparatus and slowly to stir, regulate pH to 8.0, then mouse-anti rubella virus monoclonal antibody 0.5mg is added by every 100ml colloidal gold solution 0.5mg antibody, continue stir 30min, then add final concentration be 1% BSA close, continue stir 30min.After mark terminates, carry out centrifugal with 10000r/min to marking fluid, abandon supernatant, then precipitation redissolution solution is carried out redissolving to original volume.The formula of described redissolution solution is: Tris1.965g, Casein-Na2.5g, trisodium citrate 2.5g, Tween-20 0.50ml, sucrose 10.0g, and Sodium azide 1.0g, is settled to 1L with ultrapure water.The golden label solution of having redissolved is spread 18cm by every ml solution
2ratio uniform paving on the glass fibers, be placed in 37 DEG C, relative humidity, lower than in the environment of 20%, dry 12 ~ 18 hours, makes label pad.
3, the preparation of antigen pad
Used by the rubella virus antigen of acquisition antigen protection liquid to be diluted to 8 times and obtain antigen liquid, the antigen liquid diluted is spread 18cm by every ml solution
2ratio uniform paving on the glass fibers, be placed in 37 DEG C, relative humidity, lower than in the environment of 40%, dry 4 hours, makes antigen pad.The formula of described antigen protection liquid is: Na
2hPO
45.72g, NaH
2pO
40.624g, trisodium citrate 8.5g, sucrose 10g, Sodium azide 1g, purified water is settled to 1L.
4, the preparation of coated film
Use 0.02M, pH be 7.2 phosphate buffer mouse-anti human IgG antibody (T line solution) is diluted to 1.5mg/ml, sheep anti-mouse igg (C line solution) is diluted to 1.0mg/ml.Then use a stroke film instrument to be coated on NC film by T line and C line solution equably by the liquid outlet quantity of 1.5ul/cm, NC film sticks on plastic bottom board in advance, after bag is done, NC film is placed in 37 DEG C, relative humidity, lower than in the environment of 40%, dry 3 ~ 4 hours, makes coated film.
5, the assembling of rubella virus IgG antibody Test paper
(temperature 20 ~ 25 DEG C in dry environments, relative humidity is lower than 30%), coated film is placed in clean environment, at the T line end of NC film, label pad (pad in advance cutting becomes 6mm wide) is crimping NC film 2mm closely, antigen pad (antigen pad in advance cutting becomes 10mm wide) is crimping label pad opposite side 2mm closely, and sample pad closely crimps antigen pad opposite side 4 ~ 5mm, finally with cutting cutter, the base plate posted is cut into the wide test strips of 4mm.
6, the using method of rubella virus IgG antibody Test paper
By serum to be checked or blood plasma or whole blood (measuring samples) balance to room temperature, the above-mentioned Test paper prepared lies against on testing table, 5ul measuring samples is added at sample pad place, if containing rubella virus IgG antibody in sample, after sample adds, rubella IgG antibody first forms antigen-rubella virus IgG antibody compound with rubella antigens in antigen pad, compound moves forward, the mouse-anti rubella monoclonal antibody marked with the gold in label pad forms the mouse-anti rubella monoclonal antibody immunity compound that rubella IgG antibody-rubella antigens-Jin marks, this compound moves forward along paper slip due to chromatography effect, the mouse-anti rubella monoclonal antibody immunity compound forming mouse-anti human IgG antibody-people's rubella IgG antibody-rubella antigens-Jin and mark is reacted to detection T line and mouse-anti human IgG antibody, this compound can be gathered in detection zone, when the compound assembled reaches certain quantity, then form a macroscopic band (T line), judged result is positive, if do not contain in sample or containing the rubella virus IgG antibody seldom measured, then can not form immune complex, then can not form macroscopic band, judged result is negative.C line is as the quality control standard of reagent, and positive and negative detection sample all can produce band.With the naked eye directly observe the outlet situation of test strips in 20 minutes, and judge testing result.
The minimal protection titre of the rubella virus IgG antibody 7, specified according to the World Health Organization (WHO) is 10IU/ml, when rubella virus IgG antibody concentration >=10IU/ml illustrates that human body has repellence to rubella virus, when rubella virus IgG antibody concentration < 10IU/ml illustrates that human body does not also produce repellence to rubella virus, therefore the detection critical value detecting rubella virus IgG antibody is set to 10IU/ml by this test strips, T line shows concentration >=10IU/ml that line shows rubella virus IgG antibody, and nettle rash vaccine is inoculated successfully; The not aobvious line of T line shows the concentration < 10IU/ml of rubella virus IgG antibody, and nettle rash vaccine is not inoculated successfully.
8, clinical sample testing result contrast
Use Test paper described in the application and the commercial ELISA kit that detects to detect to the clinical serum sample that company collects simultaneously, then testing result is contrasted.
Take ELISA kit as contrast, rubella virus IgG antibody is detected, detects 192 parts of rubella IgG positive sample, 108 parts of rubella IgG negative sample, and the application's ELISA test strip goes out 191 parts of rubella IgG positive sample, 109 parts of rubella IgG negative sample, overall coincidence rate is 99.33%.Clinical detection result shows, the application's Test paper performance compare without significant difference with ELISA kit, illustrate that kit described in the utility model is suitable for clinical examination, there is practical value.
Obviously, above-described embodiment is only for the example done clearly is described, and the restriction not to embodiment.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here exhaustive without the need to also giving all embodiments.And therefore amplified apparent change or variation are still within the protection domain of the utility model creation.
Claims (5)
1. nettle rash vaccine effect of inoculation Fast Evaluation kit, comprise box body, container and the Test paper of sample diluting liquid are housed, described sample diluting liquid is housed container and Test paper be positioned at box body, it is characterized in that, described Test paper is by base plate and mutually closely overlaps the sample pad be pasted onto on base plate, antigen pad, label pad, coated film and absorption pad composition, described antigen pad is coated with rubella antigens, described label pad is coated with the mouse-anti rubella monoclonal antibody of colloid gold label, described coated film is fixed with and detects T line and Quality Control C line, described detection T line is coated with mouse-anti human IgG antibody, described Quality Control C line is coated with sheep anti-mouse igg antibody, the lowest detection line of described Test paper is 10IU/ml.
2. kit according to claim 1, is characterized in that, described antigen pad is glass fibre, nonwoven fabrics or polyester film.
3. kit according to claim 1, is characterized in that, described base plate is PVC base plate, and the one side of described PVC base plate is double faced adhesive tape.
4. kit according to claim 1, is characterized in that, described coated film is nitrocellulose filter.
5. kit according to claim 1, is characterized in that, described label pad and sample pad are glass fibre, and described absorption pad is thieving paper.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201420636037.9U CN204203235U (en) | 2014-10-29 | 2014-10-29 | Nettle rash vaccine effect of inoculation Fast Evaluation kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201420636037.9U CN204203235U (en) | 2014-10-29 | 2014-10-29 | Nettle rash vaccine effect of inoculation Fast Evaluation kit |
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| CN204203235U true CN204203235U (en) | 2015-03-11 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2585246C1 (en) * | 2015-06-15 | 2016-05-27 | Федеральное бюджетное учреждение науки "Московский научно-исследовательский институт эпидемиологии и микробиологии им. Г.Н. Габричевского" Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека | Method for differential diagnosis of primary and secondary immune response to rubella virus |
-
2014
- 2014-10-29 CN CN201420636037.9U patent/CN204203235U/en not_active Expired - Lifetime
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2585246C1 (en) * | 2015-06-15 | 2016-05-27 | Федеральное бюджетное учреждение науки "Московский научно-исследовательский институт эпидемиологии и микробиологии им. Г.Н. Габричевского" Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека | Method for differential diagnosis of primary and secondary immune response to rubella virus |
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