CN202794189U - Quick whole-course quantitative immunochromatographic detection kit for C-reactive protein - Google Patents
Quick whole-course quantitative immunochromatographic detection kit for C-reactive protein Download PDFInfo
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- CN202794189U CN202794189U CN 201220399463 CN201220399463U CN202794189U CN 202794189 U CN202794189 U CN 202794189U CN 201220399463 CN201220399463 CN 201220399463 CN 201220399463 U CN201220399463 U CN 201220399463U CN 202794189 U CN202794189 U CN 202794189U
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Abstract
The utility model discloses a quick whole-course quantitative immunochromatographic detection kit for C-reactive protein. The kit is characterized by consisting of a test paper card, a colorimetric card and an immunochromatographic result interpretation recording instrument. The test paper card adopt the structure as follows: a sample loading pad (2) is adhered to one end of a plastic bottom plate (1); one end of the sample loading pad is in tight pressure connection with a colloidal gold pad (3) containing a specific antibody marking anti-human C-reactive protein (CRP); one end of the colloidal gold pad is in tight pressure connection with a nitrocellulose (NC) membrane (4); the NC membrane is coated by a detection line T (5) and a quality control line C (6) which are separated mutually; the line T is a CRP antibody paired with labeled antibody; the line C is an anti-mouse IgG antibody; the other end of the NC membrane is connected with a sample absorbing pad (7) to form test paper; and the test paper is placed into a plastic card to form the test paper card. During detection, the immunochromatographic result interpretation recording instrument or the colorimetric card is used for performing quantitative or semiquantitative judgment on the result, so the kit has the advantages of simplicity, convenience and quickness in operation, suitability for on-site detection, economy, practicability and the like.
Description
Technical field
The utility model relates to the biologic applications technical field, particularly relates to the quick omnidistance quantitatively immunochromatographytest test kit of a kind of c reactive protein with the colloidal gold immunity chromatography preparation.
Background technology
Human in the acute stage of tissue damage, some synthetic plasma proteinss of liver significantly increase, and these protein are commonly referred to as acute phase protein, and c reactive protein (CRP) is that variation is the most significant a kind of in the acute phase protein.
CRP its content in normal human serum is atomic; Sustain damage at tissue, CRP can sharply rise within a few hours when inflammation, infection or tumor destruction, CRP is widely used in early diagnosis and the antidiastole of clinical disease, its rising is found in: 1, tissue damage, infection, tumour, miocardial infarction and a series of active chronic inflammation disease, such as rheumatic arthritis, systemic vasculitis, polymyalgia rheumatism; 2, the index of postoperative infection and complication: patients after surgery CRP raises, and the CRP level should descend in postoperative 7-10 days, does not reduce or again rising prompting possibility accompanying infection or thromboembolism such as CRP; 3, can be used as the antidiastole of bacterial infection and viral infection: most of bacterial infections can cause that patients serum CRP raises, and viral infection then majority does not raise.
It is also closely related with following generation angiocardiopathy that Recent study is sent out the CRP level, a large amount of research datas show, atheroma also is a chronic inflammation process, and the slight rising of CRP is relevant with coronary artery events, apoplexy and peripheral angiopathy, is independently hazards; The probability that acute apoplexy occurs high quick CRP (Hs-CRP) level rising person is 2 times of normal healthy people, and the probability that myocardial infarction occurs is 3 times of normal person.European hypertension prevention and control guide (ESH/ESC) formal recommendation in 2003, the hyperpietic need detect the hs-CRP level.
Domestic immune turbidimetry reagent C RP for full automatic biochemical apparatus has occurred two kinds at present: higher linearity is arranged common CRP but sensitivity is bad, and higher sensitivity is arranged high quick CRP but linearity is lower, and purposes is to some extent difference also; In recent years, along with the development of inspection technology, begin to have occurred a kind of omnidistance CRP, this CRP can satisfy higher sensitivity, can satisfy higher linearity again, and purposes is wider.But omnidistance CRP detects reagent and mainly adopts chemoluminescence method or emulsion process preparation, is only suitable for serum or plasma sample, is not suitable for whole blood sample.The omnidistance CRP detection kit that adopts the preparation of immunochromatography colloidal gold technique is not yet arranged.
The immunochromatography colloidal gold technique is novel diagnostic techniques, ultimate principle is as follows: utilize a kind of antigen of colloid gold label or antibody, at the coated corresponding pairing antigen of the NC of reagent film 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, then chromatography on the NC film, again coated antigen or antibody capture, form macroscopic detection T line, under certain conditions, power and the concentration in the sample of T line are proportionate or negative correlation, thereby realize detecting.Have easy and simple to handle, reaction fast, high, the high specificity of susceptibility, be fit to Site Detection and the advantage such as economical and practical.
Summary of the invention
The purpose of this utility model provides fast omnidistance quantitatively immunochromatographytest test kit of c reactive protein, have omnidistance quantitatively or the half-quantitative detection ability, easy and simple to handle, reaction fast, high, the high specificity of susceptibility, be fit to Site Detection and the advantage such as economical and practical.
C reactive protein is omnidistance quantitatively immunochromatographytest test kit fast, it is characterized in that kit by test card, colorimetric card and immunochromatography as a result the interpretation registering instrument consist of.
Described test card structure is: paste loading pad (2) at plastic bottom board (1) one end, the tight crimping of one end of loading pad contains the collaurum pad (3) of the anti-human c reactive protein specific antibody of mark, the tight crimping cellulose nitrate of collaurum pad one end NC film (4), be coated with the detection line T (5) and the nature controlling line C (6) that are separated from each other on the NC film, the CRP antibody that detection line is and labelled antibody matches, nature controlling line is dynamics, the other end of nitrocellulose filter connects to be inhaled sample pad (7) and forms test paper, and test paper is packed into and formed test card in the plastic clip.
The antibody of described test card is high quick CRP pairing antibody, identifies respectively CRP and identifies different epi-positions, and antibody can be monoclonal antibody or the polyclonal antibody of purifying.
The NC film of described test card is the porous spline structure film of aperture 8-12 micron, and the loading pad is glass fibre membrane or nonwoven fabrics, inhales the sample pad and is made of absorbent filter.
Described colorimetric card is the cardboard that is printed on the red lines of the serial different depth on the white background, is used for the contrast of testing result is judged.Totally 6 in the red lines of the different depths, respectively the CRP concentration of correspondence 1,3,10,20,50,200mg/L.
Described immunochromatography as a result interpretation registering instrument is a kind of Systems for optical inspection, is used for the quantitative judgement to testing result, is 0-200mg/L to the sensing range of CRP.
Described test card, the coated process of NC membrane antibody is: the solution that CRP antibody is mixed with 0.6-2mg/ml with 0.01M pH7.2 phosphate buffer (PBS), anti-mouse IgG is mixed with the solution of 1-2mg/ml, rule respectively with the parameter of 1-1.5ul/cm in NC film upper and lower with spray film instrument, coated C, T line, after the line with the NC film at drying room, temperature 20-25 ℃, humidity is less than 30%, dry 2-5 hour.
Described test card, collaurum pad preparation process is: prepare diameter as the colloidal gold solution of 30-50nm take gold chloride-trisodium citrate reduction method, get 100ml collaurum liquid after preparation is finished and be placed in the beaker, use 0.2M K
2CO
3Transfer to pH8.0, press another strain CRP antibody that the 100ml colloidal gold solution adds 0.5-2mg and coated CRP pairing, stirring at room 2 hours, adding final concentration is 0.5% bovine serum albumin(BSA), 0.5% PEG 20000 sealing 20min, centrifugal 30 minutes of 12000r/m abandons supernatant, redissolve to 50ml with the collaurum working fluid, press 1ml solution and spread 20cm
2Ratio be layered on equably on glass fibre membrane or the nonwoven fabrics, put again drying room, temperature 20-25 ℃, humidity dry 2-5 hour, is made the collaurum pad less than 30%.
The assembling process of described test card is: in hothouse, temperature 20-25 ℃, humidity is less than 40%, get plastic bottom board, paste at the middle part that coated NC film is placed on plastic bottom board, and the collaurum pad is cut into suitable width, at NC film T line one side overlap joint collaurum pad, take 1/5 of collaurum pad and paste, paste the loading pad at collaurum pad opposite side overlap joint, take 1/3 of collaurum pad and paste; Inhale the sample pad at NC film C line one side overlap joint, take 1/10 stickup of inhaling the sample pad; With cutter the plastic plate that posts is cut into the wide test strips of 3-5mm at last, reinstalls in the plastic clip, form the reagent test strip card.
Described kit, detection method is: 1) will detect reagent and sample balance to room temperature, and take out test card, and keep flat; 2) accurately draw 10 μ l serum, plasma sample, draw the 20ul sample when sample is whole blood, add in the clean centrifuge tube, add again 500ul sample dilution (physiological saline or PBS), fully mixing; 3) sample of drawing after the 10ul dilution with liquid-transfering gun joins in the sample aperture, adds immediately 100 μ L sample dilutions in the damping fluid hole of bottom again, in 15-20 minute with immunochromatography as a result interpretation registering instrument or colorimetric card (partly) quantitative judgement result; When 4) instrument is judged, set and test card is put into the storehouse behind the instrument correlation parameter and detect, instrument will demonstrate the quantitative measurement result of sample concentration; During 5) with the colorimetric card result of determination, the shade of standard lines on the color of test card T line and the colorimetric card is compared, sxemiquantitative judges that the concentration of sample is interval.
The beneficial effects of the utility model are: a kind of quick omnidistance quantitatively immunochromatographytest test kit of c reactive protein that utilizes the preparation of immunochromatography colloidal gold technique is provided, quantitatively sensing range is wide, comprise the mensuration interval that Gao Min and common CRP measure kit, be fit to simultaneously serum, blood plasma and whole blood sample, and be fit to clinically single part of detection.Have easy and simple to handle, reaction fast, high, the high specificity of susceptibility, be fit to Site Detection and the advantage such as economical and practical.
Description of drawings:
Fig. 1 is the fast omnidistance quantitatively test card of immunochromatographytest test kit of c reactive protein
The reference numeral explanation:
1: plastic bottom board; 2: the loading pad; 3: the collaurum pad; The 4:NC film;
5: detection line T; 6: nature controlling line C; 7: inhale the sample pad
Embodiment
Embodiment: c reactive protein is the preparation of omnidistance quantitatively immunochromatographytest test kit and detection fast
1 main material
1.1CRP specificity pairing antibody, CRP standard items: Finland MEDIX limit company product; Mouse-anti human IgG antibody: U.S. Arista company product; Gold chloride: Sigma company product; Cellulose nitrate (NC) film: Millipore company product; Bovine serum albumin(BSA) (BSA), polyglycol PEG20000, caseinhydrolysate: Sigma product.Other common agents is analytical reagent.
1.2 clinical sample is obtained in relevant hospital by company, totally 200 parts, wherein serum sample is 100 parts, each 50 parts of blood plasma and whole blood samples, and CRP content distribution interval is the definite value serum between the 0.1-300mg/L.
1.3 the immunochromatography result ties the interpretation registering instrument: model: NS001, Newscen Coast Bio-Pharmaceutical Co., Ltd.'s product.
2 methods
2.1CRP antibody colloidal gold mark gold chloride one trisodium citrate reduction method prepares the colloidal gold solution that diameter is 30-40nm, gets three parts of collaurums after preparation is finished, and uses respectively 0.2M K
2CO
3Solution is transferred to pH7.5, pH8.0 and pH8.5.Then solution is placed on the magnetic stirring apparatus and slowly stir, slowly be added drop-wise in colloidal gold solution with CRP antibody mark by the every adding of 100ml solution 0.5mg, 1mg, 1.5mg, continue to stir 2 hours, join final concentration again and be 0.5% PEG2000 and 0.5% BSA and seal 20min, mark is centrifugal with 12000r/m after finishing, and abandons supernatant, precipitation is redissolved (pH8.0 to the collaurum working fluid of different proportionings by 50% original volume, contain BSA, sheep blood serum, sucrose and surfactant).Then the mark colloidal gold solution is pressed 1ml solution and spread 20cm
2The ratio application of sample on nonwoven fabrics, at temperature 20-25 ℃, relative humidity is made the collaurum pad dry 2-5 hour of＜30% drying room.
2.2NC film is coated will to be coated with the CRP antibody dilution with 0.01M pH7.2PBS and to become 0.6mg/ml, 1mg/ml, 1.5mg/ml, the mouse-anti human IgG is diluted to respectively 1mg/ml, 2mg/ml, then on the NC film, respectively rule coated by 1.2ul/cm with spray film instrument, after coated the finishing with the NC film at temperature 20-25 ℃, relative humidity was dry 2-5 hour of＜30% drying room.
2.3 test card is assembled in the hothouse, temperature 20-25 ℃, humidity is less than 40%, get and mould base plate, paste at the middle part that coated NC film is placed on plastic bottom board, and the collaurum pad is cut into suitable width, at NC film T line one side overlap joint collaurum pad, take 1/5 of collaurum pad and paste, paste the loading pad at collaurum pad opposite side overlap joint, take 1/3 of collaurum pad and paste; Inhale the sample pad at NC film C line one side overlap joint, take 1/10 stickup of inhaling the sample pad; To post plastic plate with cutter at last and be cut into the wide test strips of 3-5mm, reinstall in the plastic clip, form test card.
2.4 test paper technological parameter debugging with concentration not isolabeling, coated reagent make up pairing, the preparation sample utilizes the CRP standard items that reagent is tested, and seeks best of breed.
After 2.5 colorimetric card preparation and the setting of instrument parameter of curve determine the test paper technological parameter, respectively with 1,3,10,20,50, the CRP standard items of 200mg/L measure test paper, the standard items of variable concentrations demonstrate the varying strength colour band, the colour band of respective strengths is printed onto on the colorimetric card, finishes the colorimetric card preparation; Input immunochromatography behind the color band digital of respective strengths as a result in the interpretation registering instrument, is finished the setting of instrument parameter of curve.
2.6 detection method 1) will detect reagent and sample balance to room temperature, take out test card, keep flat; 2) accurately draw 10 μ l serum, plasma sample, draw the 20ul sample when sample is whole blood, add in the clean centrifuge tube, add again 500ul sample dilution (PBS), fully mixing; 3) sample of drawing after 10ul dilutes with liquid-transfering gun joins in the sample aperture, in the damping fluid hole of bottom, add immediately again 100 μ L sample dilutions (PBS), in 15-20 minute with immunochromatography as a result interpretation registering instrument or colorimetric card (partly) quantitative judgement result; When 4) instrument is judged, set and test card is put into the storehouse behind the instrument correlation parameter and detect, instrument will demonstrate the quantitative measurement result of sample concentration; During 5) with the colorimetric card result of determination, the shade of standard lines on the color of test card T line and the colorimetric card is compared, sxemiquantitative judges that the concentration of sample is interval.
After 2.7 the preparation of clinical sample detection reagent is finished, by detection method all clinical samples are detected, and analyzing and testing result.
3 results
3.1 the test paper parameter is determined the testing result according to sample, the optimum mark pH value of having determined test paper is 8.0; CRP antibody optimum mark amount is the 1mg/100ml colloidal gold solution; Best collaurum working fluid is the 20mM borate buffer solution, and pH8.0 contains 0.5%BSA, 10% sheep blood serum, 2% sucrose, 0.2%Tween20; Best bag CRP antibody concentration is 1.5mg/ml.The optimal decision time of testing result is 15-20 minute.But above parameter may need suitable adjustment when preparation different batches product.
3.2 clinical sample detects 200 parts of clinical definite value pattern detection, when quantitatively judging as a result with instrument, 195 duplicate samples mean deviation values in sensing range are all less than 15%, and maximum deviation is less than 20%, R2＞0.97, consistency coefficient＞0.88.During with the colorimetric card result of determination, the coincidence rate of interval result of determination and sample value is 95.3%, consistency coefficient＞0.85.Testing result shows that the detection kit of preparation is functional, is suitable for clinical detection, satisfies the different differentiation needs that detect occasion of different clients.
Claims (6)
1. the quick omnidistance quantitatively immunochromatographytest test kit of a c reactive protein, it is characterized in that kit by test card, colorimetric card and immunochromatography as a result the interpretation registering instrument consist of.
2. kit according to claim 1, it is characterized in that the test card structure is: paste loading pad (2) at plastic bottom board (1) one end, the tight crimping of one end of loading pad contains the collaurum pad (3) of the anti-human c reactive protein of mark (CRP) specific antibody, the tight crimping cellulose nitrate of collaurum pad one end NC film (4), be coated with the detection line T (5) and the nature controlling line C (6) that are separated from each other on the NC film, the CRP antibody that detection line is and labelled antibody matches, nature controlling line is dynamics, the other end of nitrocellulose filter connects to be inhaled sample pad (7) and forms test paper, and test paper is packed into and formed test card in the plastic clip.
According to claim 1 with 2 described kits, the antibody that it is characterized in that described test card is high quick CRR pairing antibody, identifies respectively CRP and identifies different epi-positions, antibody can be monoclonal antibody or the polyclonal antibody of purifying.
According to claim 1 with 2 described kits, the NC film that it is characterized in that described test card is the porous spline structure film of aperture 8-12 micron, the loading pad is glass fibre membrane or nonwoven fabrics, inhales the sample pad and is made of absorbent filter.
5. kit according to claim 1 is characterized in that described colorimetric card is the cardboard that is printed on the red lines of the serial different depth on the white background, is used for the contrast of testing result is judged.Totally 6 in the red lines of the different depths, respectively the CRP concentration of correspondence 1,3,10,20,50,200mg/L.
6. kit according to claim 1, it is characterized in that immunochromatography as a result the interpretation registering instrument be a kind of Systems for optical inspection, be used for the quantitative judgement to testing result, be 0-200mg/L to the sensing range of CRP.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103543272A (en) * | 2013-10-17 | 2014-01-29 | 天津中新科炬生物制药有限公司 | Rapid and quantitative detection device and method for simultaneously detecting heart-type fatty acid-binding protein and cardiac troponin I |
| CN104198731A (en) * | 2014-08-28 | 2014-12-10 | 宁波瑞源生物科技有限公司 | C-reactive protein (CRP) semi-quantitative detection reagent and test paper using reagent |
| CN104897905A (en) * | 2015-06-01 | 2015-09-09 | 上海凯创生物技术有限公司 | C reactive protein colloidal gold detection kit |
| CN105424932A (en) * | 2015-12-30 | 2016-03-23 | 天津中新科炬生物制药有限公司 | Detection device for simultaneously and quantitatively detecting SAA/PCT/CRP |
| CN105467135A (en) * | 2015-12-30 | 2016-04-06 | 天津中新科炬生物制药有限公司 | Manufacturing method of detection device for quantitatively detecting SAA/PCT/CRP at same time |
| CN105572394A (en) * | 2016-01-20 | 2016-05-11 | 上海奥普生物医药有限公司 | Preparation method of urine HCG quantitative detection test paper strip |
| CN106749659A (en) * | 2017-01-19 | 2017-05-31 | 江苏众红生物工程创药研究院有限公司 | A kind of anti-human CRP antibody and its application |
| CN108226511A (en) * | 2016-12-14 | 2018-06-29 | 北京金华科生物技术有限公司 | A kind of novel saliva CRP Test papers and preparation method thereof |
| CN112485446A (en) * | 2020-11-18 | 2021-03-12 | 重庆中元汇吉生物技术有限公司 | Kit for measuring full-range C-reactive protein and preparation method thereof |
| CN112710844A (en) * | 2020-12-16 | 2021-04-27 | 北京开景基因技术有限公司 | Semi-quantitative kit and method for detecting novel coronavirus neutralizing antibody |
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2012
- 2012-08-07 CN CN 201220399463 patent/CN202794189U/en not_active Expired - Lifetime
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103543272A (en) * | 2013-10-17 | 2014-01-29 | 天津中新科炬生物制药有限公司 | Rapid and quantitative detection device and method for simultaneously detecting heart-type fatty acid-binding protein and cardiac troponin I |
| CN104198731A (en) * | 2014-08-28 | 2014-12-10 | 宁波瑞源生物科技有限公司 | C-reactive protein (CRP) semi-quantitative detection reagent and test paper using reagent |
| CN104897905A (en) * | 2015-06-01 | 2015-09-09 | 上海凯创生物技术有限公司 | C reactive protein colloidal gold detection kit |
| CN105424932A (en) * | 2015-12-30 | 2016-03-23 | 天津中新科炬生物制药有限公司 | Detection device for simultaneously and quantitatively detecting SAA/PCT/CRP |
| CN105467135A (en) * | 2015-12-30 | 2016-04-06 | 天津中新科炬生物制药有限公司 | Manufacturing method of detection device for quantitatively detecting SAA/PCT/CRP at same time |
| CN105572394A (en) * | 2016-01-20 | 2016-05-11 | 上海奥普生物医药有限公司 | Preparation method of urine HCG quantitative detection test paper strip |
| CN108226511A (en) * | 2016-12-14 | 2018-06-29 | 北京金华科生物技术有限公司 | A kind of novel saliva CRP Test papers and preparation method thereof |
| CN106749659A (en) * | 2017-01-19 | 2017-05-31 | 江苏众红生物工程创药研究院有限公司 | A kind of anti-human CRP antibody and its application |
| CN106749659B (en) * | 2017-01-19 | 2020-05-08 | 江苏众红生物工程创药研究院有限公司 | Anti-human CRP antibody and application thereof |
| CN112485446A (en) * | 2020-11-18 | 2021-03-12 | 重庆中元汇吉生物技术有限公司 | Kit for measuring full-range C-reactive protein and preparation method thereof |
| CN112710844A (en) * | 2020-12-16 | 2021-04-27 | 北京开景基因技术有限公司 | Semi-quantitative kit and method for detecting novel coronavirus neutralizing antibody |
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Address after: 300457 Tianjin Development Zone, Sixth Street, No. 65 Patentee after: NEWSCEN COAST BIO PHARMACEUTICAL CO.,LTD. Address before: 300457 Tianjin Development Zone, Sixth Street, No. 65 Patentee before: Tianjin NewScen Coast Bio-Pharmaceutical Co.,Ltd. |
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Granted publication date: 20130313 |