CN201965131U - Quick test paper of hepatitis A virus (HAV) IgM antibody - Google Patents
Quick test paper of hepatitis A virus (HAV) IgM antibody Download PDFInfo
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- CN201965131U CN201965131U CN2010206658394U CN201020665839U CN201965131U CN 201965131 U CN201965131 U CN 201965131U CN 2010206658394 U CN2010206658394 U CN 2010206658394U CN 201020665839 U CN201020665839 U CN 201020665839U CN 201965131 U CN201965131 U CN 201965131U
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- 108091007433 antigens Proteins 0.000 claims abstract description 24
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Images
Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The utility model relates to the technical field of biological application, in particular to a quick test paper of a hepatitis A virus (HAV) IgM antibody, which comprises an upper sample pad, a colloidal gold pad which is marked with an HAV antibody, an HAV antigen pad, a nitrocellulose (NC) membrane and a sample absorption pad; the NC membrane is coated with a test (T) line and a quality control (C) line which are separated from each other; and an indicating line is sprayed between the C line and the T line. The T line is an anti-human IgM antibody, and the C line is a goat anti-mouse IgG antibody. When the quick test paper of the HAV IgM antibody is used for testing, a sample is added on the indicating line, dilution liquid is added on the upper sample pad, and then the immune reaction results are observed. The quick test paper of the HAV IgM antibody has the advantages of convenience in operation, quick reaction, high sensitivity, strong specificity, applicability to on-site test, economy, practicality 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 hepatitis A virus (HAV) IgM antibody test paper.
Background technology
Hepatitis A is a kind of of epidemic hepatitis, because infecting, hepatitis A virus (HAV) causes, and be that a class is acute, self-limited disease, the prognosis bona is seldom changed into chronic.HAV propagates by fecal oral route, promptly by hepatitis A virus polluted source, food, apparatus and life in latent period of patient or acute stage ight soil, the blood closely the contact per os enter intestines and stomach and propagate.This virus infections can cause acute symptom in various degree, comprises explosive hepatitis.The early spring in 1988, the precipitate hepatitis A that Shanghai takes place is very popular, and has totally continued 2 months, and hepatitis A the infected surpasses 350,000 people, dead 31 people.The early diagnosis of hepatitis A is significant.
Anti-HAV-IgM is the specific index of diagnosis first type viral hepatitis.Anti--HAV IgM positive means that the patient is in the acute stage of viral hepatitis type A, and can be used as the reliability index of early diagnosis.
The immunochromatography colloidal gold technique is a kind of novel diagnostic techniques, obtained comparatively extensively using, its ultimate principle is for utilizing a kind of antigen of colloid gold label or antibody, bag is matched antigen or antibody accordingly on the NC of reagent film, 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 line, have or not the judgement of realization the result by detection line.Have easy and simple to handle, advantages such as reaction is quick, susceptibility is high, high specificity, suitable on-the-spot detection.
Summary of the invention
The purpose of this utility model provides a kind of have easy, accurate, quick, sensitivity and specificity height, economical and practical, the antihepatitis A virus IgM quick detection test paper that is suitable for characteristics such as on-the-spot detection.
Take following technical scheme in order to reach the purpose of this utility model: a kind of antihepatitis A virus IgM quick detection test paper, pasting the sample pad on the plastic support board, one end of last sample pad closely connects the collaurum pad that contains underlined HAV antibody, the close-connected antigen pad that contains HAV antigen of the other end of collaurum pad, the other end of antigen pad closely connects nitrocellulose filter, nitrocellulose filter is coated with detection line and the nature controlling line that is separated from each other, detection line is the anti-human IgM antibody that is coated on the nitrocellulose filter, nature controlling line is the sheep anti-mouse igg antibody that is coated on the nitrocellulose filter, drawing between detection line and nature controlling line and apart from detection line 2.5～3mm place has index line to be used to identify the application of sample position, the other end of nitrocellulose filter closely connects to be inhaled the sample pad and forms test strips, and test strips also can be packed into and be formed the cassette packing in the plastic clip.
Described index line application of sample amount is 3～5ul.
Described upward sample pad is glass fibre membrane or nonwoven fabrics, and inhaling the sample pad is absorbent filter.
Described antihepatitis A virus IgM quick detection test paper, the method for coating that it is characterized in that described anti-human IgM antibody is: will resist human IgM antibody to be mixed with the solution of 1.5mg/ml with 0.01MpH7.2 phosphate buffer (PBS), there is Membrane jetter to rule with the parameter of 1.3ul/cm in NC film bottom, bag is by the T line, wrap by sheep anti-mouse igg on NC film top simultaneously as the C line, after the line with NC film dry 8～10 hours at drying room (20～25 ℃ of temperature, humidity is less than 30%).
Described antihepatitis A virus IgM quick detection test paper, the method that it is characterized in that described HAV antibody labeling 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 finished and be placed in the beaker, use 0.2MK
2CO
3Transfer to pH8.2, press the 100ml colloidal gold solution and add the 0.5mgHAV monoclonal antibody, stirring at room 2 hours, add final concentration 0.1% casein-sodium, 0.1% Macrogol 2000,0 sealing 20min, 12000 left the heart 30 minutes, abandoned supernatant, redissolve to 66.7ml with the collaurum working fluid, press 1ml solution shop 16cm
2Ratio be layered on equably on glass fibre membrane or the nonwoven fabrics, put 20～25 ℃ of drying room temperature again,, humidity is made the collaurum pad less than 30% drying 2～4 hours, and is standby.
Described antihepatitis A virus IgM quick detection test paper is characterized in that described HAV antigen pad preparation method for HAV antigen colloidal gold working fluid is diluted by (1: 4), presses 1ml solution shop 18.5cm
2Ratio be layered on equably on glass fibre membrane or the nonwoven fabrics, put 20～25 ℃ of drying room temperature again, humidity is made the antigen pad less than 30% drying 2～4 hours, and is standby.
The assembly method of test strips is: 20～25 ℃ of dry indoor temperatures, humidity is less than 30%, get plastic support board, paste at the middle part that the NC film that wraps quilt is placed on plastic support board, overlap the antigen pad in NC film T line one side apart from edge 1mm, opposite side at antigen overlaps the collaurum pad apart from edge 1mm, pastes the sample pad at collaurum pad opposite side apart from edge 2mm overlap joint; Inhale the sample pad in NC film C line one side apart from edge 1.5mm place overlap joint.With cutter the plastic plate that posts is cut into 3mm or the wide test strips of 4mm then.The test strips that cuts can reinstall in the plastic clip, forms HAV IgM antibody test reagent card.
Detection method: with tested serum or blood plasma balance to room temperature, the test strips or the reagent card that prepare are kept flat, on NC film sign line, add 3～5ul test sample, if containing anti-HAV IgM antibody in the sample is then caught by the anti-human IgM antibody on the NC film, and combine with the Ag of Ag-Ab-HAV-collaurum on being diffused into the NC film, form compound, when captive colloidal gold composite reaches some, then form a macroscopic 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 20 minutes with the naked eye, and judgement testing result.
The advantage of the utility model antihepatitis A virus IgM quick detection test paper: during detection sample is added on the index line 8, on last sample pad, adds dilution, observe the immune response result then.Have easy, accurate, quick, sensitivity and specificity height, economical and practical, be suitable for characteristics such as on-the-spot detection.
Description of drawings
Fig. 1 is the utility model antihepatitis A virus IgM quick detection test paper structural representation.
1: go up sample pad 2: the collaurum pad
3: antigen pad 4: nitrocellulose filter
5: inhale sample pad 6: detection line
7: nature controlling line 8: index line
9: plastic support board
Embodiment
Embodiment prepares the antihepatitis A virus IgM quick detection test paper:
1 main material
HAV native antigen, monoclonal antibody, anti-μ chain monoclonal antibody is the Shenzhen City Fapon Biotech Co., Ltd provides, and is respectively applied for hapten pad, mark colloid gold particle, bag by cellulose nitrate (NC) film; Gold chloride and trisodium citrate are Sigma company product, are used for blank gold preparation; Nitrocellulose filter is a Millipore company product; Casein-sodium, Macrogol 2000 0 are Sigma company product, are used for the collaurum sealing; Other reagent is analytical reagent.
2 methods
2.1HAV the mark of monoclonal antibody
Prepare the colloidal gold solution that diameter is 40nm with gold chloride-trisodium citrate reduction method, get three parts of collaurums after preparation is finished, use 0.2MK respectively
2CO
3PH value of solution can be transferred to pH7.0, pH8.2, pH9.2, solution be placed on the magnetic stirring apparatus and slowly stir then, slowly be added drop-wise in the colloidal gold solution by every 100ml solution 0.5mg, 1.0mg, 1.5mgHAV monoclonal antibody, continue to stir 2 hours, adding final concentration again is 0.1% casein-sodium, 0.1% Macrogol 2000 0 sealed 20 minutes, and 12000 left the heart 30 minutes, abandoned supernatant, redissolve to 66.7ml with the collaurum working fluid, press 1ml solution shop 16cm
2Ratio be layered on equably on glass fibre membrane or the nonwoven fabrics, put 20～25 ℃ of drying room temperature again,, humidity is made the collaurum pad less than 30% drying 2～4 hours, and is standby.
2.2HAV antigen pad preparation method
HAV antigen is diluted by 1: 1,1: 2,1: 4,1: 8 with the collaurum working fluid, press 1ml solution shop 18.5cm
2Ratio be layered on equably on glass fibre membrane or the nonwoven fabrics, put 20～25 ℃ of drying room temperature again, humidity is made the antigen pad less than 30% drying 2～4 hours, and is standby.
2.3 anti-μ chain monoclonal antibody bag quilt
Be diluted to 1.0mg/ml, 1.5mg/ml, 2.0mg/ml respectively with the anti-μ chain monoclonal antibody of 0.01M pH7.2PBS bag, to resist the μ chain monoclonal antibody on the NC film, to wrap quilt with spray film instrument then by the 1.3ul/cm line, wrap simultaneously by sheep anti-mouse igg antibody, after bag is done the NC film was placed drying room dry 8-10 hour, standby.
2.4 the assembling of test strips
20～25 ℃ of dry indoor temperatures, humidity is less than 30%, getting plastic support analyses, paste at the middle part that the NC film that wraps quilt is placed on plastic support board, overlap the antigen pad in NC film T line one side apart from edge 1mm, opposite side at antigen overlaps the collaurum pad apart from edge 1mm, pastes the sample pad at collaurum pad opposite side apart from edge 2mm overlap joint; Inhale the sample pad in NC film C line one side apart from edge 1.5mm place overlap joint.With cutter the plastic plate that posts is cut into 3mm or the wide test strips of 4mm then.The test strips that cuts can reinstall in the plastic clip, forms HAV IgM antibody test reagent card.
2.4 detection method: with tested serum or blood plasma balance to room temperature, the test strips or the reagent card that prepare are kept flat, on NC film sign line, add 3～5ul test sample, if containing anti-HAV IgM antibody in the sample is then caught by the anti-human IgM antibody on the NC film, and combine with the Ag of Ag-Ab-HAV on being diffused into the NC film, form compound, when captive colloidal gold composite reaches some, then form a macroscopic 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 20 minutes with the naked eye, and judgement testing result.
3 results
3.1 testing result according to sample, the optimum mark pH value of determining test paper is 8.2, the optimum mark amount is the every 100ml colloidal gold solution of 0.5mg, best collaurum working fluid is the Tris-citrate buffer solution of pH9.0, contains 0.2% casein, 5% sucrose, 0.2% BSA, 0.2% tween, best anti-μ chain monoclonal antibody bag is 1.5mg/ml by concentration, best antigen diluent ratio is 1: 4.Sentence read result in 20 minutes.
3.2 clinical sample detects
With ELISA reagent is contrast, and 20 parts of positive serums, this reagent detect positive 19 parts, and sensitivity is in 95%, 50 part of negative sample, and this reagent detects negative 49 parts, and specificity is 98%.
Claims (3)
1. antihepatitis A virus IgM quick detection test paper, it is characterized in that on plastic support board (9), pasting sample pad (1), one end of last sample pad (1) closely connects the collaurum pad (2) that contains underlined HAV antibody, the close-connected antigen pad (3) that contains HAV antigen of the other end of collaurum pad (2), the other end of antigen pad (3) closely connects nitrocellulose filter (4), nitrocellulose filter (4) is coated with detection line (6) and the nature controlling line (7) that is separated from each other, detection line (6) is for being coated on the anti-human IgM antibody on the nitrocellulose filter, nature controlling line (7) is for being coated on the sheep anti-mouse igg antibody on the nitrocellulose filter, drawing between detection line (6) and nature controlling line (7) and apart from detection line (6) 2.5～3mm places has index line (8) to be used to identify the application of sample position, the other end of nitrocellulose filter (4) closely connects to be inhaled sample pad (5) and forms test strips, and test strips also can be packed into and be formed the cassette packing in the plastic clip.
2. antihepatitis A virus IgM quick detection test paper according to claim 1 is characterized in that described index line (8) application of sample amount is 3～5ul.
3. antihepatitis A virus IgM quick detection test paper according to claim 1 is characterized in that described upward sample pad (1) is glass fibre membrane or nonwoven fabrics, and inhaling sample pad (5) is absorbent filter.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010206658394U CN201965131U (en) | 2010-12-17 | 2010-12-17 | Quick test paper of hepatitis A virus (HAV) IgM antibody |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010206658394U CN201965131U (en) | 2010-12-17 | 2010-12-17 | Quick test paper of hepatitis A virus (HAV) IgM antibody |
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| Publication Number | Publication Date |
|---|---|
| CN201965131U true CN201965131U (en) | 2011-09-07 |
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| CN2010206658394U Expired - Lifetime CN201965131U (en) | 2010-12-17 | 2010-12-17 | Quick test paper of hepatitis A virus (HAV) IgM antibody |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102608313A (en) * | 2012-02-27 | 2012-07-25 | 中国疾病预防控制中心病毒病预防控制所 | Anti-hepatitis A virus IgM (immunoglobulin M) AlphaLISA detection kit |
| CN102680694A (en) * | 2012-05-24 | 2012-09-19 | 蓝十字生物药业（北京）有限公司 | Colloidal gold test strip for rapid detection of Human enterovirus 71 (EV71) IgM (immunoglobulin m) |
| CN102692502A (en) * | 2012-04-26 | 2012-09-26 | 北京北方生物技术研究所 | Rapid hepatitis A virus IgM antibody detection kit and preparation method thereof |
| CN103336119A (en) * | 2013-06-27 | 2013-10-02 | 潍坊市康华生物技术有限公司 | Anti-HAV (hepatitis A virus) colloidal gold test kit |
| CN103364561A (en) * | 2013-07-29 | 2013-10-23 | 武汉中博生物股份有限公司 | Porcine circovirus 2 (PCV2) antibody colloidal gold immunity chromatography detection test paper and making method thereof |
| CN103399154A (en) * | 2013-07-29 | 2013-11-20 | 武汉中博生物股份有限公司 | Test paper for colloidal gold immunochromatograohic assay of IgG (immunoglobulin G) antibody of dog anti-rabies virus and preparation method of test paper |
| CN103969440A (en) * | 2014-05-27 | 2014-08-06 | 武汉中博生物股份有限公司 | Porcine reproductive and respiratory syndrome virus (PRRSV) IgM antibody colloidal gold immunochromatographic assay test paper, and preparation method and application thereof |
| CN103969451A (en) * | 2014-05-27 | 2014-08-06 | 武汉中博生物股份有限公司 | Porcine circovirus type 2 (PCV2) IgM antibody colloidal gold immunochromatographic assay test paper, and preparation method and application thereof |
| CN103983781A (en) * | 2014-05-27 | 2014-08-13 | 武汉中博生物股份有限公司 | Porcine pseudorabies virus gE IgM antibody colloidal gold immunochromatograohic assay test strip as well as preparation method and application thereof |
| 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 |
| CN104515850A (en) * | 2013-09-30 | 2015-04-15 | 万冰 | Quick test device and quick test method |
| CN116087534A (en) * | 2022-12-30 | 2023-05-09 | 天津德祥生物技术股份有限公司 | Erythrocyte antigen detection method and detection kit |
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2010
- 2010-12-17 CN CN2010206658394U patent/CN201965131U/en not_active Expired - Lifetime
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102608313A (en) * | 2012-02-27 | 2012-07-25 | 中国疾病预防控制中心病毒病预防控制所 | Anti-hepatitis A virus IgM (immunoglobulin M) AlphaLISA detection kit |
| CN102692502B (en) * | 2012-04-26 | 2015-02-04 | 北京北方生物技术研究所 | Rapid hepatitis A virus IgM antibody detection kit and preparation method thereof |
| CN102692502A (en) * | 2012-04-26 | 2012-09-26 | 北京北方生物技术研究所 | Rapid hepatitis A virus IgM antibody detection kit and preparation method thereof |
| CN102680694A (en) * | 2012-05-24 | 2012-09-19 | 蓝十字生物药业（北京）有限公司 | Colloidal gold test strip for rapid detection of Human enterovirus 71 (EV71) IgM (immunoglobulin m) |
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