CN101949926A - Human echinococcosis colloidal gold immunochromatographic assay urine testing quick diagnosis test paper card - Google Patents
Human echinococcosis colloidal gold immunochromatographic assay urine testing quick diagnosis test paper card Download PDFInfo
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- CN101949926A CN101949926A CN201010244078XA CN201010244078A CN101949926A CN 101949926 A CN101949926 A CN 101949926A CN 201010244078X A CN201010244078X A CN 201010244078XA CN 201010244078 A CN201010244078 A CN 201010244078A CN 101949926 A CN101949926 A CN 101949926A
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Abstract
The invention discloses a human echinococcosis colloidal gold immunochromatographic assay urine testing quick diagnosis test paper card. The test paper card comprises a plastic card box and a detection module in the card box; the plastic card box is provided a sample loading hole and an observation window; the detection module comprises sample filtering paper, a nitric acid fibrous membrane and absorbent paper; echinococcosis resistance fluid antigen humanized single chain variable region antibody which is marked by colloidal gold is absorbed on the absorbent paper and nitric acid fibrous membrane; a quality control line and a detection line are marked on the echinococcosis resistance fluid antigen humanized single chain variable region antibody marked by the colloidal gold on nitric acid fibrous membrane; the sample filtering paper is opposite to the sample loading hole; and the nitric acid fibrous membrane is opposite to the observation window. The test paper card has the advantages of simplicity and convenience, fastness, no invasiveness, reliability and stability.
Description
Technical field
The present invention relates to the medical biotechnology field, particularly a kind of Urine test paper of quick diagnosis human body echinococcosis can utilize the observable change color diagnosis of naked eyes whether to infect echinococcosis fast.
Background technology
Echinococcosis has the onset concealment as a kind of zoonosis of serious harm human health, and slower development recurs frequent characteristics, needs early diagnosis and detection regularly.Popular serious at China's zoonosis, the Ministry of Public Health proposes the target of " to reaching district occurred frequently 60% population coverage in the echinococcosis epidemiology survey task " in April, 2007.Community hospital, epidemic prevention station, medical colleges and schools and scientific research institution need special scientific research apparatus that this disease is carried out serology, immunology and imaging diagnosis but the field investigation of popular district and grass-roots unit lack special instrument and equipment personnel, cause a lot of cases can not early find early to diagnose early treatment.
Summary of the invention
The objective of the invention is to overcome weak point of the prior art, the present invention is based on the immune response device that colloidal gold immunochromatographimethod detects principle, a kind of human body echinococcosis colloidal gold immunochromatographimethod examination urine fast diagnose test paper card is provided.
A kind of human body echinococcosis colloidal gold immunochromatographimethod examination urine fast diagnose test paper card of the present invention, it comprises the detection module in plastics cartridge and the cartridge; On the described plastics cartridge well and view window are arranged; Described detection module comprises filter sample paper, nitrocellulose membrane and thieving paper; Be adsorbed with strand variable region, the anti-echinococcosis cyst fluid antigen people source antibody of colloid gold label on described thieving paper and the nitrocellulose membrane; Colloid gold label on the described nitrocellulose membrane has on the antibody of strand variable region, anti-echinococcosis cyst fluid antigen people source and is marked with a nature controlling line and a detection line; Described filter sample paper is facing to well, and described nitrocellulose filter is facing to view window.
Described view window is a rectangle.
Described detection line is by marked by streptavidin.
Described nature controlling line is by the rabbit anti-mouse igg mark.
The preparation of described collaurum and mark may further comprise the steps: tri-distilled water dissolved chlorine auric acid, be heated to boiling, to final concentration be 0.01%, every 100ml adds 10% trisodium citrate aqueous solution 1.5ml; Boil 10min again and be orange red, use 0.2mol/LK after the cooling
2CO
3Transfer to pH8.5, stir the anti-echinococcosis cyst fluid antigen people source strand variable region antibody 2mg that adds purifying down fast, the protein 25 0mg that adds cow's serum, stir 10min fast, add 10%NaCl, the concentration that makes NaCl is 1%, shake up, respectively with 2000,4000, the 10000r/min gradient centrifugation, 10min/ time, obtain the antibody conjugates that sediment is the golden mark of preliminary purification at last, be dissolved in storage liquid, the antibody conjugates of golden mark is strand variable region, the anti-echinococcosis cyst fluid antigen people source antibody of golden mark.
The mark of described nitrocellulose filter may further comprise the steps: the nitrocellulose filter bag is by strand variable region, the anti-echinococcosis cyst fluid antigen people source antibody of colloid gold label, and mark respectively is wide 1: 100 Streptavidin of 1mm and rabbit anti-mouse igg on the nitrocellulose filter; Dry the back with the 0.1mol/LpH7.4PBS sealing that contains 1% NBCS, be bonded at successively on the white plastic sheet after the drying, be cut into 0.6cm * 10cm bar ,-4 ℃ store for future use.
The present invention has following advantage:
Utilize the Protocols in Molecular Biology development to have easy (without instrument and equipment, do not need the professional and technical personnel to operate, can use in patient family), fast (finishing analysiss within 1-5 minute), do not have to create and (in urine, test, do not need collecting blood sample), the reliable external quick diagnosis reagent kit of the human body echinococcosis of (but quantitative interpretation result, diagnostic accordance rate reaches more than 90%), stable (product is being stablized more than a year under the regulation preservation condition).The specificity of Echinococcus hydatid cyst antibody and susceptibility have determined the sensitivity and the specificity of this diagnostic tool.Employing, can reduce the cross reaction with other diseases, thereby improve sensitivity and specificity as the antibody of catching Echinococcus hydatid cyst antigen at strand variable region, the people source antibody of the reorganization holoprotein of echinococcosis cyst fluid antigen B, has reduced and has failed to pinpoint a disease in diagnosis or mistaken diagnosis.
Description of drawings
Fig. 1 is the structural representation of embodiments of the invention.
Fig. 2 is the external structure synoptic diagram of Fig. 1.
Embodiment
The present invention is further described by the following examples:
With reference to figure 1, Fig. 2, pick-up unit: be made up of the plastics cartridge of outside and the detection module of the inside, outside plastics cartridge comprises base 1 and shell 2, and well 3 and rectangle view window 4 are arranged on the shell 2; Inner detection module is made up of filter sample paper 5, nitrocellulose membrane 6, thieving paper 7.Strand variable region, the anti-echinococcosis cyst fluid antigen people source antibody 8 of colloid gold label is adsorbed on thieving paper 7 and the nitrocellulose membrane 6, forms a nature controlling line 9 and a detection line 10; Filter sample paper 5 is over against well 3, and nitrocellulose membrane 6 is over against view window 4; Detection line 10 is by marked by streptavidin; Nature controlling line 9 is by the rabbit anti-mouse igg mark.
Testing process: urine sample enters behind the well because capillarity is mobile to the thieving paper direction, middle through bag in advance by the zone of strand variable region, anti-echinococcosis cyst fluid antigen people source antibody of colloid gold label, golden label is redissolved fully, antigen and golden labelled antibody form golden labelled antibody-antigenic compound, continue to flow to forward on the nitrocellulose membrane, be fixed with the detection line place of capture antigen.When the result is positive, on golden labelled antibody-antigenic compound, will form golden labelled antibody-antigen-insolubilized antibody compound, present red positive band; As there not being antigen in the sample, then be not hunted down, just do not develop the color, then the result is negative.Unnecessary free gold labelled antibody continues to move forward to and is fixed with anti-golden marked body two anti-places, and being fixed presents red nature controlling line.
Make raw material: gold chloride: Beijing chemical reagent factory; Streptavidin: BNHS:Sigma; Strand variable region, anti-echinococcosis cyst fluid antigen people source antibody: (GENE BANK:AF407670), provide by Zhejiang University, can purchase, or Zhejiang University provides in Zhejiang University; Nitrocellulose membrane: Sigma; Rabbit anti-mouse igg, Sigma.
Preparation of Colloidal Gold and mark: tri-distilled water dissolved chlorine auric acid, be heated to boiling, to final concentration be 0.01%, every 100ml adds 10% trisodium citrate aqueous solution 1.5ml.Boil 10min again and be orange red, use 0.2mol/LK after the cooling
2CO
3Transfer to pH8.5, stir the anti-echinococcosis cyst fluid antigen people source strand variable region antibody 2mg that adds purifying down fast, add the protein 25 0mg of cow's serum, stir 10min fast, add 10%NaCl, the concentration that makes NaCl is 1%, shake up, respectively with 2000,4000, the 10000r/min gradient centrifugation, 10min/ time, obtain the antibody conjugates that sediment is the golden mark of preliminary purification at last, be dissolved in storage liquid.
The cellulose nitrate membrane marker: the nitrocellulose membrane bag is by strand variable region, the anti-echinococcosis cyst fluid antigen people source antibody of colloid gold label, 1: 100 Streptavidin (detection line) of mark and rabbit anti-mouse igg (control line) on the nitrocellulose membrane, and it is wide to be 1mm.Drying the back seals with the 0.1mol/L pH7.4PBS that contains 1% NBCS.Be bonded at successively on the white plastic sheet after the drying.Be cut into 0.6cm * 10cm bar ,-4 ℃ store for future use.
The result judges: 1 redness of position T appearance with detection line on the nitrocellulose membrane is the Echinococcus hydatid cyst positive; If have only the nature controlling line redness negative; Article two, line does not all have colour developing and is test-strips inefficacy.
At last, it is also to be noted that what more than enumerate only is specific embodiments of the invention.Obviously, the invention is not restricted to above examples of implementation, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.
Claims (6)
1. a human body echinococcosis colloidal gold immunochromatographimethod tries urine fast diagnose test paper card, and it is characterized in that: it comprises the detection module in plastics cartridge and the cartridge; On the described plastics cartridge well and view window are arranged; Described detection module comprises filter sample paper, nitrocellulose membrane and thieving paper; Be adsorbed with strand variable region, the anti-echinococcosis cyst fluid antigen people source antibody of colloid gold label on described thieving paper and the nitrocellulose membrane; Colloid gold label on the described nitrocellulose membrane has on the antibody of strand variable region, anti-echinococcosis cyst fluid antigen people source and is marked with a nature controlling line and a detection line; Described filter sample paper is facing to well, and described nitrocellulose filter is facing to view window.
2. according to the described test card of claim 1, it is characterized in that: described view window is a rectangle.
3. according to the described test card of claim 1, it is characterized in that: described detection line is by marked by streptavidin.
4. according to the described test card of claim 1, it is characterized in that: described nature controlling line is by the rabbit anti-mouse igg mark.
5. the preparation of the described collaurum of claim 1 and mark is characterized in that may further comprise the steps: tri-distilled water dissolved chlorine auric acid, be heated to boiling, to final concentration be 0.01%, every 100ml adds 10% trisodium citrate aqueous solution 1.5ml; Boil 10min again and be orange red, use 0.2mol/LK after the cooling
2CO
3Transfer to pH8.5, stir the anti-echinococcosis cyst fluid antigen people source strand variable region antibody 2mg that adds purifying down fast, the protein 25 0mg that adds cow's serum, stir 10min fast, add 10%NaCl, the concentration that makes NaCl is 1%, shake up, respectively with 2000,4000, the 10000r/min gradient centrifugation, 10min/ time, obtain the antibody conjugates that sediment is the golden mark of preliminary purification at last, be dissolved in storage liquid, the antibody conjugates of golden mark is strand variable region, the anti-echinococcosis cyst fluid antigen people source antibody of golden mark.
6. the mark of the described nitrocellulose filter of claim 1, it is characterized in that may further comprise the steps: the nitrocellulose filter bag is by strand variable region, the anti-echinococcosis cyst fluid antigen people source antibody of colloid gold label, and mark respectively is wide 1: 100 Streptavidin of 1mm and rabbit anti-mouse igg on the nitrocellulose filter; Dry the back with the 0.1mol/L pH7.4PBS sealing that contains 1% NBCS, be bonded at successively on the white plastic sheet after the drying, be cut into 0.6cm * 10cm bar ,-4 ℃ store for future use.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102608330A (en) * | 2012-03-12 | 2012-07-25 | 中国疾病预防控制中心寄生虫病预防控制所 | Immunochromatographic strip for detecting alveolar echinococcosis and preparation method thereof |
| CN103995121A (en) * | 2014-02-21 | 2014-08-20 | 大连大学 | Preparation method for colloidal-gold test paper based on single-chain antibody |
| CN107402298A (en) * | 2016-05-18 | 2017-11-28 | 上海新吉而生物科技有限公司 | A kind of dog excrement echinococcosis antigen quick detection kit and its application method |
| CN107884578A (en) * | 2017-11-01 | 2018-04-06 | 杭州微瑞科技有限公司 | For echinococcosis antibody test card in Quantitative detection serum |
| CN108181473A (en) * | 2017-12-25 | 2018-06-19 | 迈克生物股份有限公司 | Schistosome antibody detection kit |
| CN108535477A (en) * | 2018-04-11 | 2018-09-14 | 中国疾病预防控制中心寄生虫病预防控制所 | A kind of rapid detection method of echinococcosis intermediate host lesion internal organs |
| CN111521816A (en) * | 2020-04-21 | 2020-08-11 | 沈阳农业大学 | Time-resolved fluorescence immunochromatographic assay test strip for echinococcosis granulosus of cattle and preparation method thereof |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102608330A (en) * | 2012-03-12 | 2012-07-25 | 中国疾病预防控制中心寄生虫病预防控制所 | Immunochromatographic strip for detecting alveolar echinococcosis and preparation method thereof |
| CN103995121A (en) * | 2014-02-21 | 2014-08-20 | 大连大学 | Preparation method for colloidal-gold test paper based on single-chain antibody |
| CN103995121B (en) * | 2014-02-21 | 2015-11-18 | 大连大学 | Based on the preparation method of the gold test strip of single-chain antibody |
| CN107402298A (en) * | 2016-05-18 | 2017-11-28 | 上海新吉而生物科技有限公司 | A kind of dog excrement echinococcosis antigen quick detection kit and its application method |
| CN107884578A (en) * | 2017-11-01 | 2018-04-06 | 杭州微瑞科技有限公司 | For echinococcosis antibody test card in Quantitative detection serum |
| CN108181473A (en) * | 2017-12-25 | 2018-06-19 | 迈克生物股份有限公司 | Schistosome antibody detection kit |
| CN108535477A (en) * | 2018-04-11 | 2018-09-14 | 中国疾病预防控制中心寄生虫病预防控制所 | A kind of rapid detection method of echinococcosis intermediate host lesion internal organs |
| CN111521816A (en) * | 2020-04-21 | 2020-08-11 | 沈阳农业大学 | Time-resolved fluorescence immunochromatographic assay test strip for echinococcosis granulosus of cattle and preparation method thereof |
| CN111521816B (en) * | 2020-04-21 | 2023-03-21 | 沈阳农业大学 | Time-resolved fluorescence immunochromatographic assay test strip for echinococcosis granulosus of cattle and preparation method thereof |
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Application publication date: 20110119 |