CN105181960A - Fluorescent immunochromatography test paper and preparation method thereof - Google Patents
Fluorescent immunochromatography test paper and preparation method thereof Download PDFInfo
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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Abstract
The invention discloses fluorescent immunochromatography test paper and a preparation method thereof. The test paper is formed by sequentially overlapping a sample pad (1), an antibody carrying film (2) and absorbent paper (3) through a base plate (4) which is provided with binding agents, the antibody carrying film (2) is coated with a marker mixture joint line (5), a testing line (6) and a quality control line (7) in sequence from the end close to the sample pad (1), and the marker mixture joint line (5) is formed by arranging a marker mixture obtained by coupling fluorescent microspheres with an antibody 1 and biotin within a pretreatment area (8) which is pretreated with the antibody carrying film in a coating mode through a crossed film metal-spraying instrument. By means of the test paper, the problems that the result is not accurate caused by insufficient release amount due to the fact that excessive binding materials remain on glass fibers are solved, the tested fluorescence value is raised, and the sensitivity or detection upper limit is lifted. Meanwhile, in the production, the antibody carrying film is coated with the marker mixture obtained by coupling the fluorescent microspheres with the antibody 1 and the biotin, the marker mixture joint line, the T line and the C line can be coated simultaneously, time and processes are saved, and the production efficiency is improved.
Description
Technical field
The present invention relates to Test paper field, particularly relate to a kind of fluorescence immune chromatography Test paper and preparation method.
Background technology
Test paper major part used is clinically made up of 4 parts, namely formed by there being the base plate of bonding agent to overlap mutually successively successively by sample pad, pad, antibody carrier film, thieving paper successively, or sample pad directly uses glass fibre membrane, saves pad.Wherein, pad to be coated with near antibody carrier film side or to spray with fluorescent microsphere respectively with the potpourri of antibody 1 and biotin coupling label.On antibody carrier film, bag is detected T line and the Quality Control C line of the antibody (or antigen) of project, namely Cleaning Principle adds sample to sample pad, nitride layer to be checked in sample analyses joint line, compound is formed after fluorescent microsphere on thing to be checked and joint line and the potpourri specific reaction of antibody 1 coupling label, chromatography is to T line, form double antibodies sandwich or competition, biotinylated derivative chromatography on joint line forms specific reaction to C line and Avidin, fluorescent quantitation detection system is utilized to detect the fluorescence intensity of the enrichment in T line and C line region, the content of determinand in reflected sample.
This method has 3 shortcomings: 1, due to glass fibre membrane material factors, fluorescent microsphere can only be sprayed with the potpourri of antibody 1 and biotin coupling label respectively, or soak, if spray, have uneven shortcoming, affect precision, be not suitable for quantitative detection; If soak, not only uneven and a large amount of microballoons and antibody can be wasted, increase cost; 2, the label potpourri part be sprayed on glass fibre membrane can hook the solid space of glass fibre element film, cause each chromatography homogeneity poor, all have label potpourri to be in various degree trapped on glass fibre membrane at every turn, make preci-sion and accuracy become large.3, the homogeneity of spray membrane process by technique, causes the homogeneity of antibody-fluorescent bond poor less than bag.
Summary of the invention
The object of the present invention is to provide a kind of raising fluorescent value, precision, save material and the fluorescence immune chromatography Test paper of Simplified flowsheet.
For achieving the above object, the invention provides a kind of fluorescence immune chromatography Test paper, comprise sample pad (1), antibody carrier film (2), thieving paper (3), overlapped mutually by the base plate (4) with bonding agent successively and form, from wrapping the potpourri joint line (5) being labeled thing respectively successively near sample pad (1) one end on described antibody carrier film (2), detection line (6) and nature controlling line (7), it is characterized in that, the potpourri joint line (5) of described label is coated in the pretreated pretreatment zone of antibody carrier film (8) with drawing film gold spraying instrument with the potpourri of antibody 1 and biotin coupling label respectively by fluorescent microsphere.
Further, the pre-service of described antibody carrier film is using stroke film gold spraying instrument bag by the pretreatment fluid of upper width 1.8-2.2mm near one end of sample pad (1); Preferably, near sample pad (1) one end and apart from edge 4mm antibody carrier film (2) film on draw a film gold spraying instrument bag quilt; Described pretreatment fluid is that the 0.01MPBS of PH7.4 adds 20 quality volume % sucrose, 2 quality volume %BSA and 0.02 quality volume %NaN
3;
Further, the potpourri joint line (5) of described label is positioned at the middle position of the pretreated pretreatment zone of antibody carrier film (8).
Further, described detection line (6) bag is by the antibody 2 of detected project or antigen.
Further, potpourri joint line (5) width of the width greater than flag thing of described pretreatment zone (8); Preferably, the width of pretreatment zone (8) is 2 times of potpourri joint line (5) width of label.
Further, described fluorescent microsphere adopts rare earth elements europium or fluorescein to be wrapped on silica dioxide granule or latex particle; Preferably, described fluorescent microsphere is wrapped in silica dioxide granule for adopting rare earth elements europium or fluorescein.
Further, the potpourri joint line (5) of described label is positioned near sample pad (1) one end and apart from antibody carrier film (2) film of edge 4mm, the potpourri joint line (5) of label and detection line (6) are separated by potpourri joint line (5) distance nature controlling line (7) 15mm of 10mm, label.
On the other hand, the present invention also provides a kind of preparation method of described fluorescence immune chromatography Test paper, it is characterized in that, step is,
Pretreatment fluid prepare: get PH be 7.4 0.01MPBS solution add 20 quality volume % sucrose, add 2 mass volume ratio %BSA and 0.02 quality volume %NaN
3, fully after mixing;
The preparation of the potpourri of label: after selecting the amino of fluorescent microsphere and the aldehyde radical coupling of glucosan, 1:1 adds antibody 1 and mixes in mass ratio, on glucosan, other aldehyde radicals with the amino coupled on antibody 1, can form the potpourri of fluorescent microsphere and antibody 1 coupling label; Preferably, select the 1:1 mixing in mass ratio of fluorescent microsphere and antibody 1, form the potpourri of fluorescent microsphere and antibody 1 coupling label; In kind, after the amino of fluorescent microsphere and the aldehyde radical coupling of glucosan, 1:1 adds biotin mixing in mass ratio, and on glucosan, other aldehyde radicals with the amino coupled on biotin, can form the label of fluorescent microsphere and biotin coupling; By the 1:1 mixing by volume of described two kinds of labels, and with containing the pH7.4 of 1 mass ratio %BSA, 0.01MPBS500 times of dilution mixture thing;
The preparation of T line coating buffer: the monoclonal antibody of mouse-anti test item is diluted to working concentration with pH7.4,0.01MPBS dilution;
The preparation of C line coating buffer: Avidin is diluted to working concentration with pH7.4,0.01MPBS dilution;
The pre-service of antibody carrier film (2): select nitrocellulose membrane as antibody carrier film, near the one end will pasting glass fibre membrane with draw film gold spraying instrument bag by width be about the above-mentioned pretreatment fluid prepared of 2mm, the region that this width is about 2mm is called pretreatment zone; 45 DEG C of 2h are dried;
Bag is by line: wrapped with the potpourri fluorescence of antibody 1 and biotin coupling label, T line coating buffer, C line coating buffer respectively by fluorescent microsphere by drawing film gold spraying instrument by above-mentioned pretreated nitrocellulose membrane simultaneously, form potpourri joint line, T line, the C line of label, described 3 lines, live width is 1mm; Potpourri joint line (5) and detection line (6) the distance about 10mm of label, potpourri joint line (5) and nature controlling line (7) the distance about 15mm of label; Afterwards by this bag by after NC film dry in 45 DEG C of 2h; The potpourri joint line (5) of described label is positioned at the middle position of pretreatment zone;
Be assembled into test card: by glass fibre membrane and dry bag by after nitrocellulose membrane, thieving paper paste assembling successively, assemble rear cutting cutter and be cut into the examination bar that every bar width is 4mm, put into examination bar card, pressure is blocked, and is prepared into test card.
Also on the one hand, the present invention also provides a kind of described fluorescence immune chromatography Test paper for the purposes of the detection of antigen or antibody.
Further, add sample in the sample pad of fluorescence immune chromatography Test paper described in claim 1, nitride layer to be checked in sample analyses joint line, compound is formed after antibody 1 label specific reaction on the joint line of thing to be checked and described fluorescence immune chromatography Test paper, chromatography is to T line, form double antibodies sandwich or competition, biotinylated derivative chromatography on joint line forms specific reaction to C line and Avidin, fluorescent quantitation detection system is utilized to detect the fluorescence intensity of the enrichment in T line and C line region, the content that namely typical curve draws determinand in sample is brought into by after the process of T/C fluorescence intensity level,
Preferably, the method that described typical curve obtains is, with pH7.4,0.01MPBS dilution country or international standard substance are to required linear concentration, test the corresponding fluorescence intensity level of each concentration with fluorescence immune chromatography Test paper, standard concentration and the matching of fluorescence intensity level ratio obtain related trend lines.
Fluorescence immune chromatography Test paper of the present invention be used for antigen, antibody detection, comprise human body, animals and plants, in antigen and antibody, antigen and antibody etc. in food, food additives.
Described sample pad (1) can be glass fibre element film; Base plate (4) with bonding agent can be the liner plate (PVC board) of band bonding agent.The pretreatment zone (8) at potpourri joint line (5) place of described label maintains with a certain distance from T line (6) position, ensures that pretreatment fluid does not affect T line.
Detection line of the present invention (6) is also called T line (6).
The potpourri joint line (5) of described label, the membranous part position at place is being wrapped by front through pre-service, T line (6) is detected the antibody 2 (or antigen) of project for detection line bag, C line is that nature controlling line (7) bag is by Avidin, by forming compound after the potpourri specific reaction of the fluorescent microsphere on thing to be checked and joint line and antibody 1 coupling label, chromatography is to T line (6), form double antibodies sandwich or competition, biotinylated derivative chromatography on joint line forms specific reaction to C line and Avidin, fluorescent quantitation detection system is utilized to detect the fluorescence intensity of the enrichment in T line and C line region, the content of determinand in reflected sample.
Fluorescence immune chromatography Test paper of the present invention has the efficiency of the combination of enhancing fluorescent grain microballoon and coated antibody, improves fluorescent value, improves precision, saves material, and Simplified flowsheet also compensate for the conventional advantage trying bar defect in the detection.
Script fluorescent microsphere changes into the potpourri mode be sprayed on glass fibre membrane of antibody 1 and biotin coupling label and being coated on the pretreated position of carrier film with drawing a film gold spraying instrument with the potpourri of antibody 1 and biotin coupling label respectively by fluorescent microsphere by the present invention respectively.
Fluorescence immune chromatography Test paper of the present invention avoids the following shortcoming in production:
1, the fluorescent microsphere wrapped by getting on compares evenly with the potpourri of antibody 1 and biotin coupling label respectively, and wrap by simple process, waste is few.
2, bag by fluorescent microsphere respectively with the potpourri of antibody 1 and biotin coupling label before, first wrapping by width is the pretreatment fluid of 2 times of the width that this bond is drawn, and this pretreatment fluid can play the micropore of blocking antibody carrier film, level and smooth face, forms diaphragm.After to be dried, then draw in pretreated region fluorescent microsphere respectively with the potpourri of antibody 1 and biotin coupling label, avoid bond and hook the bond that causes damage in carrier film, make directly all to discharge from pre-service face at chromatography process.Loss amount is reduced to minimum, therefore instant invention overcomes because bond is detained too much on glass, the inaccurate problem of result that burst size deficiency causes, and the fluorescent value of detection is drawn high, improves sensitivity or upper limit of detection.
3, in production, fluorescent microsphere is coated on antibody carrier film with the potpourri of antibody 1 and biotin coupling label respectively, can simultaneously by the potpourri joint line of label, T line, C line bag, time and labour saving skill.Enhance productivity.
Accompanying drawing explanation
Fig. 1 is the horizontal sectional drawing of fluorescence immune chromatography Test paper of the present invention.
Fig. 2 is the plan view from above of fluorescence immune chromatography Test paper of the present invention.
Embodiment
Be described below in detail embodiments of the invention, the example of described embodiment is shown in the drawings, and wherein same or similar label represents same or similar element or has element that is identical or similar functions from start to finish.Be exemplary below by the embodiment be described with reference to the drawings, be intended to for explaining the present invention, and can not limitation of the present invention be interpreted as.Unreceipted concrete technology or condition person in embodiment, according to the technology described by the document in this area or condition or carry out according to product description.Agents useful for same or the unreceipted production firm person of instrument, being can by the conventional products of commercial acquisition.
The structure of following examples composition graphs 1 and Fig. 2.
Embodiment 1: the preparation of fluorescence immune chromatography Test paper
A kind of fluorescence immune chromatography Test paper, overlapped mutually successively by the base plate (4) with bonding agent successively by sample pad (1), antibody carrier film (2), thieving paper (3) and form, wherein said sample pad (1) is glass fibre element film; Base plate (4) with bonding agent is the liner plate (PVC board) of band bonding agent.
Preparation method's following steps:
Prepared by pretreatment fluid: preparation PH is the 0.01MPBS solution of 7.4, for subsequent use.Get the PBS partly prepared and add 20 quality volume % sucrose, add 2 mass volume ratio %BSA and 0.02 quality volume %NaN
3, fully after mixing, 2-8 DEG C for subsequent use.
The preparation of the potpourri of label: after selecting the amino of fluorescent microsphere and the aldehyde radical coupling of glucosan, 1:1 adds antibody 1 and mixes in mass ratio, on glucosan, other aldehyde radicals with the amino coupled on antibody 1, can form the potpourri of fluorescent microsphere and antibody 1 coupling label; Preferably, select the 1:1 mixing in mass ratio of fluorescent microsphere and antibody 1, form the potpourri of fluorescent microsphere and antibody 1 coupling label; In kind, after the amino of fluorescent microsphere and the aldehyde radical coupling of glucosan, 1:1 adds biotin mixing in mass ratio, and on glucosan, other aldehyde radicals with the amino coupled on biotin, can form the label of fluorescent microsphere and biotin coupling; Both label volume ratio 1:1 are mixed, and with containing the pH7.4 of 1 mass ratio %BSA, 0.01MPBS500 times of dilution mixture thing.The preparation of T line coating buffer: with pH7.4,0.01MPBS dilution, the monoclonal antibody of mouse-anti test item is diluted to working concentration (0.5-5mg/ml adjusts according to actual experiment), 2-8 DEG C for subsequent use.
The preparation of C line coating buffer: Avidin is diluted to working concentration (0.5-5mg/ml adjusts according to actual experiment) with pH7.4,0.01MPBS dilution, 2-8 DEG C for subsequent use.
The pre-service of antibody carrier film (2): select commercially available nitrocellulose membrane and NC film as antibody carrier film, near the one end will pasting glass fibre membrane with draw film gold spraying instrument bag by width be about the above-mentioned pretreatment fluid prepared of 2mm, the region that this width is about 2mm is called pretreatment zone.45 DEG C of 2h are dried.
Bag is by line: wrapped with the potpourri of antibody 1 and biotin coupling label, T line coating buffer, C line coating buffer respectively by fluorescent microsphere by drawing film gold spraying instrument by above-mentioned pretreated NC film simultaneously, form potpourri joint line, T line, the C line of label, described 3 lines, live width is 1mm.Potpourri joint line (5) and T line (detection line 6) the distance about 10mm of label, potpourri joint line (5) and C line (nature controlling line 7) the distance about 15mm of label.Afterwards by this bag by after NC film dry in 45 DEG C of 2h.The potpourri joint line (5) of described label is positioned at the middle position of pretreatment zone;
Be assembled into test card: according to shown in Fig. 1 by the bag of glass fibre membrane and oven dry by after NC film, thieving paper paste assembling successively, assembled rear cutting cutter and be cut into the examination bar that every bar width is 4mm, put into examination bar card, pressure card, is prepared into test card.Under dry environment, lucifuge is for subsequent use.
Detection method: take out test card, after application of sample, nitride layer to be checked in sample analyses joint line, compound is formed after antibody 1 label specific reaction on the joint line of thing to be checked and described fluorescence immune chromatography Test paper, chromatography is to T line, form double antibodies sandwich or competition, the biotinylated derivative chromatography on joint line forms specific reaction to C line and Avidin, utilizes fluorescent quantitation detection system to detect the fluorescence intensity of the enrichment in T line and C line region.
Fit standard curve: be diluted to required linear concentration with the PBS of country or international standard substance 0.01MpH7.4, test the corresponding fluorescence intensity level of each concentration with fluorescence immune chromatography Test paper, standard concentration and the matching of fluorescence intensity level ratio obtain related trend lines.
Pattern detection and result: detect by above-mentioned detection method, gained fluorescence intensity level is brought in matched curve, can obtain corresponding concentration of specimens value.
The preparation of contrast test paper
Fluorescent microsphere is be sprayed on glass fibre membrane with the potpourri of antibody 1 and biotin coupling label respectively, and other all with embodiment 1, no longer describe in detail.
Embodiment 2: compliance test result
Standard curve fit: get bought progesterone standard items 0.01MpH7.4PBS and be diluted to linear concentration, and test corresponding fluorescence intensity with progesterone Test paper.
Table 1 control group and experimental group quantitative measurement standard curve data table of the present invention
Note: control group is embodiment contrast test paper; Experimental group of the present invention is the fluorescence immune chromatography Test paper of embodiment of the present invention gained.
Trendline control group y=-0.13ln (x)+0.638, R is simulated with concentration and T/C
2=0.994>=0.9801 correlativity meets standard; Experimental group y=-0.15ln (x)+0.747, R of the present invention
2=0.997>=0.9801 correlativity meets standard.
Sample: Concentration of Progesterone value is respectively 4 parts of different serum of 50ng/ml, 25ng/ml, 5ng/ml and 0ng/ml as sample;
Prepare progesterone detection kit, consistent with the preparation method of embodiment 1 fluorescence immune chromatography Test paper, its T line encrusting substance is progesterone antigen.
Method: each concentration utilizes kit to test 10 times respectively, detects and draws fluorescent value and test concentrations, and obtain the coefficient of variation and contrast.Testing result is as shown in table 1-4.Wherein experimental group is for adopting embodiment 1 gained fluorescence immune chromatography Test paper.Control group is the contrast test paper prepared adopting contrast test paper.
The Comparative result table of control group and experimental group during table 20ng/ml
The Comparative result table of control group and experimental group during table 35ng/ml
The Comparative result table of control group and experimental group during table 425ng/ml
The Comparative result table of control group and experimental group during table 550ng/ml
The above CV calculates: measure 10 results mean concentration (Mean,
), standard deviation (SD), the coefficient of variation
Wherein, X represents the measured value of some results,
represent the average measurement of n result, n represents number.
Lowest detectable limit calculates: replicate determination 10 relative fluorescence signal, calculates the mean value of relative fluorescence signal transacting value (T/C:T is detection line fluorescence signal, and C is nature controlling line fluorescence signal)
and standard deviation (SD), will
substitute into metering-response curve, the concentration value calculated is lowest detectable limit.
As can be seen from table 2-5: 1, fluorescence is analyzed, the control group result of same concentration sample and Comparative result of the present invention, the fluorescent value surveyed with the present invention is more homogeneous and higher than the fluorescent value of contrast examination, illustrate that present invention process makes the release of joint line label more thorough, low concentration is visible, improves fluorescent value and can improve sensitivity, make testing result more have clinical value.2, show through calculating, the fluorescent value CV of testing result of the present invention is less than the fluorescent value CV of control group detection paper, and can find out that test paper of the present invention obviously can reduce CV, burst size is stablized, and is a guarantee to the accuracy of clinical detection result.
In sum, the present embodiment have employed different sample range concentration as experimental subjects, experimental result be of the present invention group comparatively control group performance to have greatly improved lifting, concentration level is described, does not affect superiority of the present invention.
Involved Cleaning Principle is competition law above, but is not limited to described method.Immunochromatography sandwich method also within the scope of the invention.
Although illustrate and describe embodiments of the invention above, be understandable that, above-described embodiment is exemplary, can not be interpreted as limitation of the present invention, those of ordinary skill in the art can change above-described embodiment within the scope of the invention when not departing from principle of the present invention and aim, revising, replacing and modification.
Claims (10)
1. a fluorescence immune chromatography Test paper, comprise sample pad (1), antibody carrier film (2), thieving paper (3), overlapped mutually by the base plate (4) with bonding agent successively and form, from wrapping the potpourri joint line (5) being labeled thing respectively successively near sample pad (1) one end on described antibody carrier film (2), detection line (6) and nature controlling line (7), it is characterized in that, the potpourri joint line (5) of described label is coated in the pretreated pretreatment zone of antibody carrier film (8) with drawing film gold spraying instrument with the potpourri of antibody 1 and biotin coupling label respectively by fluorescent microsphere.
2. fluorescence immune chromatography Test paper described in claim 1, is characterized in that, the pre-service of described antibody carrier film is using stroke film gold spraying instrument bag by the pretreatment fluid of upper width 1.8-2.2mm near one end of sample pad (1); Preferably, near sample pad (1) one end and apart from edge 4mm antibody carrier film (2) film on draw a film gold spraying instrument bag quilt; Described pretreatment fluid is that the 0.01MPBS of PH7.4 adds 20 quality volume % sucrose, 2 quality volume %BSA and 0.02 quality volume %NaN
3.
3. fluorescence immune chromatography Test paper described in claim 1, is characterized in that, the potpourri joint line (5) of described label is positioned at the middle position of the pretreated pretreatment zone of antibody carrier film (8).
4. fluorescence immune chromatography Test paper described in claim 1, is characterized in that, described detection line (6) bag is by the antibody 2 of detected project or antigen.
5. fluorescence immune chromatography Test paper described in claim 1, is characterized in that, potpourri joint line (5) width of the width greater than flag thing of the potpourri joint line pretreatment zone (8) of described label; Preferably, the width of pretreatment zone (8) is 2 times of potpourri joint line (5) width of label.
6. fluorescence immune chromatography Test paper described in claim 1, is characterized in that, described fluorescent microsphere adopts rare earth elements europium or fluorescein to be wrapped on silica dioxide granule or latex particle; Preferably, described fluorescent microsphere is wrapped in silica dioxide granule for adopting rare earth elements europium or fluorescein.
7. fluorescence immune chromatography Test paper described in claim 1, it is characterized in that, the potpourri joint line (5) of described label is positioned near sample pad (1) one end and apart from antibody carrier film (2) film of edge 4mm, the potpourri joint line (5) of label and detection line (6) are separated by potpourri joint line (5) distance nature controlling line (7) 15mm of 10mm, label.
8. a preparation method for fluorescence immune chromatography Test paper described in claim 1, is characterized in that, step is,
Pretreatment fluid prepare: get PH be 7.4 0.01MPBS solution add 20 quality volume % sucrose, add 2 mass volume ratio %BSA and 0.02 quality volume %NaN
3, fully after mixing;
The preparation of the potpourri of label: after selecting the amino of fluorescent microsphere and the aldehyde radical coupling of glucosan, 1:1 adds antibody 1 and mixes in mass ratio, on glucosan, other aldehyde radicals with the amino coupled on antibody 1, can form the potpourri of fluorescent microsphere and antibody 1 coupling label; Preferably, select the 1:1 mixing in mass ratio of fluorescent microsphere and antibody 1, form the potpourri of fluorescent microsphere and antibody 1 coupling label; In kind, after the amino of fluorescent microsphere and the aldehyde radical coupling of glucosan, 1:1 adds biotin mixing in mass ratio, and on glucosan, other aldehyde radicals with the amino coupled on biotin, can form the label of fluorescent microsphere and biotin coupling; By the 1:1 mixing by volume of described two kinds of labels, and with containing the pH7.4 of 1 mass ratio %BSA, 0.01MPBS500 times of dilution mixture thing;
The preparation of T line coating buffer: the monoclonal antibody of mouse-anti test item is diluted to working concentration with pH7.4,0.01MPBS dilution;
The preparation of C line coating buffer: Avidin is diluted to working concentration with pH7.4,0.01MPBS dilution;
The pre-service of antibody carrier film (2): select nitrocellulose membrane as antibody carrier film, near the one end will pasting glass fibre membrane with draw film gold spraying instrument bag by width be about the above-mentioned pretreatment fluid prepared of 2mm, the region that this width is about 2mm is called pretreatment zone; 45 DEG C of 2h are dried;
Bag is by line: wrapped with the potpourri fluorescence of antibody 1 and biotin coupling label, T line coating buffer, C line coating buffer respectively by fluorescent microsphere by drawing film gold spraying instrument by above-mentioned pretreated nitrocellulose membrane simultaneously, form potpourri joint line, T line, the C line of label, described 3 lines, live width is 1mm; Potpourri joint line (5) and detection line (6) the distance about 10mm of label, potpourri joint line (5) and nature controlling line (7) the distance about 15mm of label; Afterwards by this bag by after NC film dry in 45 DEG C of 2h; The potpourri joint line (5) of described label is positioned at the middle position of pretreatment zone;
Be assembled into test card: by glass fibre membrane and dry bag by after nitrocellulose membrane, thieving paper paste assembling successively, assemble rear cutting cutter and be cut into the examination bar that every bar width is 4mm, put into examination bar card, pressure is blocked, and is prepared into test card.
9. fluorescence immune chromatography Test paper described in a claim 1 is used for the purposes of antigen or antibody test.
10. fluorescence immune chromatography Test paper described in claim 9 is used for the purposes of antigen or antibody test, it is characterized in that, add sample in the sample pad of fluorescence immune chromatography Test paper described in claim 1, nitride layer to be checked in sample analyses joint line, compound is formed after antibody 1 label specific reaction on the joint line of thing to be checked and described fluorescence immune chromatography Test paper, chromatography is to T line, form double antibodies sandwich or competition, biotinylated derivative chromatography on joint line forms specific reaction to C line and Avidin, fluorescent quantitation detection system is utilized to detect the fluorescence intensity of the enrichment in T line and C line region, the content that namely typical curve draws determinand in sample is brought into by after the process of T/C fluorescence intensity level,
Preferably, the method that described typical curve obtains is, with pH7.4,0.01MPBS dilution country or international standard substance are to required linear concentration, test the corresponding fluorescence intensity level of each concentration with fluorescence immune chromatography Test paper, standard concentration and the matching of fluorescence intensity level ratio obtain related trend lines.
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| CN109856407A (en) * | 2018-12-26 | 2019-06-07 | 北京勤邦生物技术有限公司 | A kind of canine distemper virus antibody fluorescence test strip and its preparation method and application |
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