CN106248974A - A kind of hypersensitivity immunity chromatography detection test paper - Google Patents
A kind of hypersensitivity immunity chromatography detection test paper Download PDFInfo
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- CN106248974A CN106248974A CN201610540862.2A CN201610540862A CN106248974A CN 106248974 A CN106248974 A CN 106248974A CN 201610540862 A CN201610540862 A CN 201610540862A CN 106248974 A CN106248974 A CN 106248974A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
- G01N33/76—Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/575—Hormones
- G01N2333/59—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
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Abstract
A kind of hypersensitivity immunity chromatography detection test paper, it is characterized in that: include base plate, and it is successively set on the sample pad on described base plate, pad, nitrocellulose filter, sample suction pad, described sample pad, described nitrocellulose filter, described sample suction pad overlap successively, constitute one for the passage that liquid stream to be measured leads to, the i.e. first horizontal channel；Its advantage is: sample reaches to detect line by the first horizontal channel, and do not combine with the traget antibody/antigen (conjugate) on pad and produce conjugate, traget antibody/antigen (conjugate) is migrated by the second horizontal channel and reaches to detect line, the dual pathways design of Test paper the most of the present invention；The dual pathways of present invention design chromatographs Avidin, the application of biotin in path and the selection passing through tracer label thing gold colloidal, cellulose nano-particle, latex particle or gold colloidal cellulose nano-particle etc. and A pad, B pad and pad; overcome the gathering/coagulation problems between analyte in conjugate and sample; enhance the ability that detection signal amplifies, thus the height that embodies the present invention is special and hypersensitivity.
Description
Technical field
The present invention relates to technical field of medical detection, specifically a kind of hypersensitivity immunity chromatography detection test paper.
Background technology
The detection method of application immunochromatography establishes its status in instant detection day by day.For different analytes
Immunochromatography colloidal gold strip, have simple to operate, detection quickly, reaction result directly perceived, can the advantage of Site Detection,
Be widely used in environmental monitoring, food inspection, field such as medical treatment detection etc. so that it become the most instant a kind of detection or from
My diagnostic means.If the detection instrument of currently used detection human chorionic gonadotropin (HCG) is blood or urine HCG gold colloidal
Immunity chromatography detection test paper, has become as the main method of the instant checkout and diagnosis of early pregnancy.
Colloidal gold immunochromatographimethod technology is to be applied to a kind of immunity mark of antigen-antibody using gold colloidal as tracer label thing
Note technology, the immunochromatography colloidal gold strip made with this technology, its weak point is that the sensitivity of detection is poor, works as sample
In as relatively low in determinand content in human blood, urine, saliva, perspiration etc. or trace time be especially apparent, during such as detection urine specimen
Result shows as false positive, and the background noise of colloidal gold strip occurs now and then, and detects saliva with colloidal gold strip equally
During liquid sample, often having false negative result to occur, the interference effect of the anti-saliva of common colloidal gold strip is poor, especially deposits in saliva
Become apparent from when a large amount of mucin；And for example haemolysis sample, when especially a large amount of haemolysis specimen colloidal gold strips detect, to inspection
Survey result has a significant impact.
In order to improve the detection sensitivity of immunochromatography colloid gold test paper, people start substantial amounts of exploration and research.Someone
The gold colloidal using oarse-grained gold colloidal, such as larger than 40nm carrys out labelling, but the poor stability of larger particles gold colloidal；For colloid
The low problem of reagent paper sensitivity of gold labelling, someone selects to apply two-sided god's granule (Janus), quantum dot etc. to substitute tracer label
Thing improves the sensitivity of detection, such as the preparation of a patent cn104655836A immuno-chromatographic test paper strip of Jiang Xingyu etc. and make
With, patent cn105004869A quantum dot immune chromatograph test strip of Wang Bing etc. and preparation method, the patent such as Hu Min
The patent cn102667482B organic coloring granule of cn203432969U carbon quantum dot test strips and Jitian dawn etc., contains
Its diagnostic medicament box and the method for in-vitro diagnosis, all highly sensitive than existing immunochromatography colloidal gold strip technology；
But these method disadvantages are background noise not to be solved, additionally need extra equipment (such as Fluorescent reader etc.),
And cost is high；Also have with different gold grain composition double combination pads in tradition chromatographic test paper, or have single two combinations
Pad, such as patent cn102507929A signal enhancement mode immuno-chromatographic test paper strip and the preparation method of Chen Wei etc., can carry
The sensitivity of high detected material, but the application of these methods also has its limitation, when in detected analyte exist interference because of
During element, in the presence of the mucin in hemocyte, saliva, antibacterial etc., sample exists or defines undesirable
Solid constituent, it has blocked labelled analyte and has led to the passage of detection zone, thus has reduced the sensitivity of detection；The most useful enzyme
Method, silver staining enhancement, magnetic bead etc. improve sensitivity, but are also to there is the problem that operation is complicated, cost is high；The one of Zhao Fuquan etc.
The test strips (patent cn204154724U) of detection sample, because label pad is covered by sample pad so that gold mark rate of release slows down,
Make the concentration proportion of analyte and gold labeling antibody in sample increase, thus improve the sensitivity of test strips；The one such as Sun Xuejiao
Highly sensitive test strips (patent cn203385721U) adds sample in sample pad, makes material to be checked directly first be combined with T line,
On buffer Sample application pad, add sample buffer again, make colloid gold label granule discharge, colloid gold label granule be combined in T line
On material to be checked combine colour developing, so on the one hand can reduce the consumption of sample, another aspect compared with traditional test strips,
If sample rests on detection line one step ahead with the antigen on detection line or antibodies, inspection can be improved to a certain extent
The sensitivity surveyed.Above by the method changing path, improve the sensitivity of detection to a certain extent, but play crucial or core
If the tracer label thing gold colloidal of heart effect does not change, the immune chromatography test paper of this single change passage, its sensitivity
Property improve limited.Above-mentioned situation shows, the most not yet has from tracer label thing, sample pad, pad, the structure of reagent paper or material
And many-sided research comprehensively optimizing immune chromatography test paper such as path and patent, if optimize immune chromatography test paper and carry comprehensively
The sensitivity of high immune chromatography test paper, it will make immunochromatography technique or detection method be more widely used.
Therefore, how to find the combination of a kind of structure by Test paper and material etc., improve reagent paper sensitivity, compel
At the eyebrows and eyelashes.
Summary of the invention
In order to solve above-mentioned technological deficiency, the present invention provides quick, simple, susceptiveness is high and a kind of Gao Min of high specificity
Perception immunity chromatography detection test paper.
In view of above-mentioned present situation, it is an object of the invention to provide one and optimize comprehensively and carry high sensitive immunity-chromatography test
Paper.It is i.e. a kind of quick, simple and result immune chromatography test paper accurately, is again when existence in the detected analyte of one
During interference factor and determinand content is relatively low or can accurately detect during trace, be simultaneously suitable for different types of body fluid (as
Blood, urine, saliva, perspiration etc.) the instant immunologic diagnosis chromatographic test paper of sample.
One hypersensitivity immunity chromatography detection test paper of the present invention, including: base plate, and be successively set on described base plate
Sample pad, pad, nitrocellulose filter, sample suction pad, described sample pad, described nitrocellulose filter, described sample suction pad depend on
Secondary overlap joint, constitutes one and is used for the passage that liquid stream to be measured is logical, be i.e. referred to as the first horizontal channel；
Wherein, also including that a barrier film, described barrier film one end are overlapped in described sample pad, the other end is overlapped on described nitric acid
On cellulose membrane, and described pad one end is overlapped on described barrier film, and the other end is overlapped on described nitrocellulose filter, institute
State and between pad, described nitrocellulose filter, described sample suction pad, constitute another passage (being referred to as the second horizontal channel), make
Dual channel mode Test paper must be constituted.
One hypersensitivity immunity chromatography detection test paper of the present invention, its advantage is: sample is reached by the first horizontal channel
Detection line, and do not combine with traget antibody/antigen (conjugate) and produce conjugate, traget antibody/antigen (conjugate) passes through
Second horizontal channel migrates and reaches to detect line, the dual pathways design of Test paper the most of the present invention；
The dual pathways of present invention design chromatographs path and by tracer label thing gold colloidal, cellulose nano-particle, breast
The selection of glue granule or gold colloidal cellulose nano-particle etc., A pad, B pad and pad, nitrocellulose filter
Middle Avidin, the application of biotin, overcome the gathering/coagulation problems between analyte in conjugate and sample, also carries simultaneously
The high excellent marker of tracer label thing, enhances the ability that detection signal amplifies, thus the height that embodies the present invention is special and
Hypersensitivity.
Accompanying drawing explanation
Fig. 1 is the first form structure schematic diagram of one hypersensitivity immunity chromatography detection test paper of the present invention；
Fig. 2,4,6 are one hypersensitivity immunity chromatography detection test paper the five, the six of the present invention, seven kind of form structure signal
Figure；
Fig. 3 is one hypersensitivity immunity chromatography detection test paper the second form structure schematic diagram of the present invention；
Fig. 5 is the third form structure schematic diagram of one hypersensitivity immunity chromatography detection test paper of the present invention；
Fig. 7 is 4th kind of form structure schematic diagram of one hypersensitivity immunity chromatography detection test paper of the present invention；
Fig. 8 is 8th kind of form structure schematic diagram of one hypersensitivity immunity chromatography detection test paper of the present invention；
Fig. 9 is the structural representation that the present invention a kind of hypersensitivity immunity chromatography detection test paper is outside equipped with shell；
Wherein:
1, base plate, 2, sample pad, 3, pad, 31, B pad, 32, A pad,
4, nitrocellulose filter, 41, detection line, 42, nature controlling line；
5, sample suction pad, 6, barrier film, 7, filter pad, 8, buffer liquid pad, 9, shell,
10, well, 11, liquid filling hole, 12, detection form, 13, sample suction pad form.
Detailed description of the invention
The first form concrete structure of Test paper of the present invention is:
A kind of hypersensitivity immunity chromatography detection test paper, including base plate 1, and is successively set on the sample on described base plate 1
Product pad 2, pad 3, nitrocellulose filter 4, sample suction pad 5, described sample pad 2, described nitrocellulose filter 4, described sample suction pad 5
Overlap successively, constitute one for the passage that liquid stream to be measured leads to, the i.e. first horizontal channel；
Wherein, also including that a barrier film 6, described barrier film 6 one end are overlapped in described sample pad 2, the other end is overlapped on described
On nitrocellulose filter 4, and described pad 3 one end is overlapped on described barrier film 6, and the other end is overlapped on described celluloid
On film 4, constituting another passage between described pad 3, described nitrocellulose filter 4, described sample suction pad 5, the i.e. second level is led to
Road so that constitute dual channel mode Test paper.(as shown in Figure 1)
Further, described sample pad 2 is about 2mm with described nitrocellulose filter 4 lap of splice.
Test paper the second form of the present invention is with the first form difference:
A filter pad 7 it is provided with, for filtering the chaff interference in detected material in described sample pad 2.Further, described
The chaff interference that filter pad 7 filters is antibacterial, mucoprotein, haemolysis, blood cell etc., and the structure in described filter pad 7 and design of material
Change according to the material filtered.(as shown in Figure 3)
The third form of Test paper of the present invention is with the first form difference:
Also it is pasted with a buffer liquid pad 8 on described barrier film 6, and described buffer liquid pad 8 one end is overlapped on described barrier film (6)
On, the other end is overlapped on described pad 3, is used for accelerating described pad 3 internal labeling thing and fully dissolves, is discharged into described nitre
On acid cellulose film 4.(as shown in Figure 5)
4th kind of form of Test paper of the present invention is with the second form difference:
Also it is pasted with a buffer liquid pad 8 on described barrier film 6, and described buffer liquid pad 8 one end is overlapped on described barrier film (6)
On, the other end is overlapped on described pad 3, is used for accelerating described pad 3 internal labeling thing and fully dissolves, is discharged into described nitre
On acid cellulose film 4.(as shown in Figure 7)
Test paper of the present invention 5th kind, the 6th kind, the 7th kind of form respectively with the first, the second, the third detection
The difference of reagent paper is all:
It is fine that described pad 3 includes that B pad 31, A pad 32, described B pad 31 one end are overlapped on described nitric acid
On dimension element film 4, described B pad 31 other end is overlapped by one end of described A pad 32, and another of described A pad 32
End is overlapped on described barrier film 6, thus constitutes double combination pad, the detection when detected material trace, improves capture ability；
(as shown in Fig. 2 or Fig. 4 or Fig. 6)
In Fig. 6, described buffer liquid pad 8 one end is overlapped on described barrier film 6, and the other end is overlapped on described A pad 32
On.
8th kind of form of Test paper of the present invention is with the 4th kind of form difference:
It is fine that described pad 3 includes that B pad 31, A pad 32, described B pad 31 one end are overlapped on described nitric acid
On dimension element film 4, described B pad 31 other end is overlapped by one end of described A pad 32, and another of described A pad 32
End is overlapped on described barrier film 6, thus constitutes double combination pad, the detection when detected material trace, improves capture ability.
(as shown in Figure 8)
In Fig. 8, described buffer liquid pad 8 one end is overlapped on described barrier film 6, and the other end is overlapped on described A pad 32
On.
Further, the 1/3-1/ of a length of described barrier film 6 total length that described buffer liquid pad 8 overlaps with described barrier film 6
4,1/4-1/5 or 2mm of a length of described pad 3 total length that described buffer liquid pad 8 overlaps with described pad 3.
Further, described nitrocellulose filter 4 is coated with detection line 41, nature controlling line 42；And described detection line 41, institute
State and on nature controlling line 42, have immobilized antigen or antibody or other ligand binding molecules, such as aptamers, nucleic acid etc..
Preferably, described pad 3 is marked with specific antibody or the gold colloidal of antigen, cellulose nano-particle, breast
Glue granule or gold colloidal cellulose nano-particle, or/and be added with Avidin on described nitrocellulose filter 4 or/and biological
Element.
Preferably, as shown in Figure 9: described Test paper also includes a shell 9, and described shell 9 includes offering successively
Well 10, liquid filling hole 11, detection form 12, sample suction pad form 13, and described well 10 is corresponding with described sample pad 2, when
When being provided with described filter pad 7 on Test paper, described well 10 is corresponding with described filter pad 7 so that sample is by described
Chromatograph after carrying out after well 10 chromatographing or pass sequentially through described filter pad 7, described sample pad 2；
Described liquid filling hole 11 is corresponding with described pad 3, when described pad 3 is divided into A pad 32, B pad 31,
Described liquid filling hole 11 is corresponding with described A pad 32, when being provided with buffer liquid pad 8 on described pad 3, and described liquid filling hole 11
Corresponding with described buffer liquid pad 8, enter described pad 3 or A pad 32 for buffer by described buffer liquid pad 8
Chromatograph after on；
Described detection form 12 with on described nitrocellulose filter 4 coated described detection line 41 right with described nature controlling line 42
Should, for observing testing result by described detection form 12；
Described sample suction pad form 13 is corresponding with described sample suction pad 5, is used for observing whether sample reaches described sample suction pad 5.
Further, described shell 9 can arrange numeral and/or alpha code, to indicate that described sample pad 2 adds sample
Described well 10, described liquid filling hole 11, described detection form 12 and the described sample suction pad form 13 etc. at place.According to by sample
Different, it is also possible to use after selecting first to be diluted by sample or mix with diluent, according to different types of sample and
Different testing sample in sample, reagent paper of the present invention may select different diluent, buffer with the use of or sample
With the mixture of buffer, and should be first by.And described pad 3 includes adding at A, B pad 31, buffer liquid pad 8
With the liquid filling hole 11 of buffer position for receiving buffer.
Described sample pad 2, described buffer liquid pad 8, described filter pad 7 are glass fibre membrane or non-woven fabrics；Described sample suction pad 5
It is made up of absorbent filter；
The present invention can be according to the different purposes of different field, and carry out that the actual selection present invention checks required for reagent paper is described
Sample pad 2, described pad 3, described nitrocellulose filter 4 and the size of described sample suction pad 5, thickness, density and material.
When the pad 3 in Test paper of the present invention is one, described pad 3 is marked with specific antibody or anti-
Former gold colloidal or cellulose nano-particle or latex particle or gold colloidal cellulose nano-particle or be added with Avidin or
Biotin, it combines with the labelling of detection line 41 on described nitrocellulose filter 4 following different modes:
1, it is marked with the gold colloidal of specific antigen or antibody on pad 3, nitrocellulose filter 4 has another special
Property antigen or antibody；
2, it is marked with the cellulose nano-particle of specific antigen or antibody on pad 3, nitrocellulose filter 4 has
Another specific antigen or antibody；
3, the gold colloidal-cellulose nano-particle of specific antigen or antibody, celluloid it are marked with on pad 3
Another specific antigen or antibody is had on film 4；
4, it is marked with gold colloidal+the biotin of specific antigen or antibody on pad 3, nitrocellulose filter 4 has
Another specific antigen or antibody+Avidin；
5, it is marked with gold colloidal+the Avidin of specific antigen or antibody on pad 3, nitrocellulose filter 4 has
Another specific antigen or antibody+biotin；
6, the gold colloidal+Avidin of specific antigen or antibody, nitrocellulose filter 4 subscript it are marked with on pad 3
Note has the gold colloidal+Avidin antibody of another specific antigen or antibody；
7, the gold colloidal+Avidin antibody of specific antigen or antibody, nitrocellulose filter 4 it are marked with on pad 3
On have another specific antigen or antibody+Avidin；
8, it is marked with gold colloidal+the biotin of specific antigen or antibody on pad 3, nitrocellulose filter 4 has
Another specific antigen or antibody+biotin antibody；
9, the gold colloidal+biotin antibody of specific antigen or antibody, nitrocellulose filter 4 it are marked with on pad 3
On have another specific antigen or antibody+biotin；
10, the cellulose nano-particle+biotin of specific antigen or antibody, cellulose nitrate it are marked with on pad 3
Another specific antigen or antibody+Avidin is had on element film 4；
11, the cellulose nano-particle+Avidin of specific antigen or antibody, cellulose nitrate it are marked with on pad 3
Another specific antigen or antibody+biotin is had on element film 4；
12, the cellulose nano-particle+Avidin of specific antigen or antibody, cellulose nitrate it are marked with on pad 3
Another specific antigen or antibody+Avidin antibody is had on element film 4；
13, the cellulose nano-particle+Avidin antibody of specific antigen or antibody, nitric acid it are marked with on pad 3
Another specific antigen or antibody+Avidin is had on cellulose membrane 4；
14, the cellulose nano-particle+biotin of specific antigen or antibody, cellulose nitrate it are marked with on pad 3
Another specific antigen or antibody+biotin antibody is had on element film 4；
15, the cellulose nano-particle+biotin antibody of specific antigen or antibody, nitric acid it are marked with on pad 3
Another specific antigen or antibody+biotin is had on cellulose membrane 4；
16, the gold colloidal-cellulose nano-particle+biotin of specific antigen or antibody, nitre it are marked with on pad 3
Another specific antigen or antibody+Avidin is had on acid cellulose film 4；
17, the gold colloidal-cellulose nano-particle+Avidin of specific antigen or antibody, nitre it are marked with on pad 3
Another specific antigen or antibody+biotin is had on acid cellulose film 4；
18, the gold colloidal-cellulose nano-particle+Avidin of specific antigen or antibody, nitre it are marked with on pad 3
Another specific antigen or antibody+Avidin antibody is had on acid cellulose film 4；
19, the gold colloidal-cellulose nano-particle+Avidin being marked with specific antigen or antibody on pad 3 resists
Body, nitrocellulose filter 4 has another specific antigen or antibody+Avidin；
20, the gold colloidal-cellulose nano-particle+biotin of specific antigen or antibody, nitre it are marked with on pad 3
Another specific antigen or antibody+biotin antibody is had on acid cellulose film 4；
21, the gold colloidal-cellulose nano-particle+biotin being marked with specific antigen or antibody on pad 3 resists
Body, nitrocellulose filter 4 has another specific antigen or antibody+biotin；
Above-mentioned various combination is equally applicable to latex particle and being mutually combined between them.
When the structure of the described pad 3 in Test paper of the present invention is divided into described A pad 32, described B pad 31
Time, described A pad 32, described B pad 31 be marked with following manner:
1, it is marked with biotinylated antibody+cellulose nano-particle on A pad 32, B pad 31 has Avidin+fibre
Dimension element nano-particle.
2, it is marked with biotinylated antibody+cellulose nano-particle on B pad 31, A pad 32 has Avidin+fibre
Dimension element nano-particle.
3, it is marked with biotinylated antibody on A pad 32, B pad 31 has Avidin+cellulose nano-particle.
4, it is marked with biotinylated antibody on B pad 31, A pad 32 has Avidin+cellulose nano-particle.
5, it is marked with biotinylated antibody+gold colloidal on A pad 32, B pad 31 has Avidin+cellulose nanometer
Granule.
6, it is marked with biotinylated antibody+gold colloidal on B pad 31, A pad 32 has Avidin+cellulose nanometer
Granule.
7, it is marked with anti-biotin antibodies+cellulose nano-particle on A pad 32, B pad 31 has biotinylation
Antibody.
8, it is marked with anti-biotin antibodies+cellulose nano-particle on A pad 32, B pad 31 has biotinylation
Antibody+cellulose nano-particle.
9, it is marked with anti-biotin antibodies+cellulose nano-particle on A pad 32, B pad 31 has biotinylation
Antibody+gold colloidal.
10, it is marked with anti-biotin antibodies+cellulose nano-particle on B pad 31, A pad 32 has biotinylation
Antibody.
11, it is marked with anti-biotin antibodies+cellulose nano-particle on B pad 31, A pad 32 has biotinylation
Antibody+cellulose nano-particle.
12, it is marked with anti-biotin antibodies+cellulose nano-particle on B pad 31, A pad 32 has biotinylation
Antibody+gold colloidal.
13, double-stranded DNA connection biotinylated antibody+cellulose nano-particle, B pad 31 it are marked with on A pad 32
On have Avidin+cellulose nano-particle.
14, double-stranded DNA connection biotinylated antibody+cellulose nano-particle, A pad 32 it are marked with on B pad 31
On have Avidin+cellulose nano-particle.
15, it is marked with biotinylated antibody+gold colloidal on A pad 32, B pad 31 has Avidin+gold colloidal.
16, it is marked with biotinylated antibody+gold colloidal on B pad 31, A pad 32 has Avidin+gold colloidal.
17, it is marked with anti-biotin antibodies+gold colloidal on A pad 32, B pad 31 has biotinylated antibody.
18, it is marked with anti-biotin antibodies+gold colloidal on B pad 31, A pad 32 has biotinylated antibody.
19, it is marked with anti-biotin antibodies+gold colloidal on A pad 32, B pad 31 has biotinylated antibody+glue
Body gold.
20, it is marked with anti-biotin antibodies+gold colloidal on B pad 31, A pad 32 has biotinylated antibody+glue
Body gold.
21, it is marked with double-stranded DNA connection biotinylated antibody+gold colloidal on A pad 32, B pad 31 has affine
Element+gold colloidal.
22, it is marked with double-stranded DNA connection biotinylated antibody+gold colloidal on B pad 31, A pad 32 has affine
Element+gold colloidal.
Above-mentioned various combination is equally applicable to latex particle and being mutually combined between them.
The present invention can be used for different types of sample, such as blood, urine, saliva, perspiration, cerebrospinal fluid etc., and can be used for surveying
The existence of what part of trial, preferably the dual pathways in the present invention ought be provided with described filter pad 7, described buffer liquid pad 8 pattern is entered
During row test, use described filter pad 7 to accept sample, reach after described sample pad 2 completes again by sample after described filter pad 7
Subsequent layers is analysed.For convenience or sample suction pad 5 described in more accurate observation has no specimen to chromatograph, described sample suction pad 5 can add coloring agent
Or painted stripes, develop the color when described sample suction pad 5 has sample to chromatograph, or can clearly be observed by described sample suction pad form 13, this
Time just can add buffer at described buffer liquid pad 8 or described pad 3, the method be better than 5 seconds-5 minutes after again combine
The mode of pad 3 addition buffer.
One hypersensitivity immunity chromatography detection test paper of the present invention, is so work:
Sample drop is added to described sample pad 2 (when for being provided with described filter pad 7, drop on described filter pad 7),
Sample by the first horizontal channel chromatography on described nitrocellulose filter 4, with the specific antigen on described detection line 41 or
Antibodies；When sample chromatography is to described sample suction pad 5, after i.e. 5 seconds-5 minutes or described sample suction pad form 13 can clearly be observed
When having sample chromatography developing, buffer is dropped to described pad 3 and (when being divided into A pad 32, B pad 31, drops to
On A pad 32；When being provided with buffer liquid pad 8 on pad 3, drop on buffer liquid pad 8) on, described pad 3 (person A
Pad 32) in label (conjugate) by second bottom horizontal flow sheet passage chromatography flow on described nitrocellulose filter 4,
Anti-in macroscopic color (male/female) with after the detectable substance specific antigen on described detection line 41 or antibodies
Should, the about several seconds to after several minutes, when there is macroscopic color in described detection line 41, described nature controlling line 42, i.e. positive detection
As a result, the antibody containing detection or antigen in detected material sample are shown, when the most described nature controlling line 42 occurs macroscopic
During color, i.e. negative result, show antibody or antigen without detection in detected material sample, when described nature controlling line 42 does not has
When macroscopic color occurs, show that reagent paper quality control goes wrong, unrelated with positive or negative testing result.
Below according to specific embodiment, the invention will be further described:
Embodiment one
The preparation of people's saliva chorionic-gonadotropin hormone (HCG) Rapid immunodiagnosis reagent paper is (when pad is one, then
The 17th kind of pattern that the labelling of pad and nitrocellulose membrane is combined as in the disclosure above)
One,
1, primary raw material is as follows:
Reagent: gold chloride, trisodium citrate, anti alpha-HCG or β-HCG antibody, cellulose nano-particle (Asahi Kasei
Company produce cellulose CNB nano-particle), BSA, trehalose, NaCl, Na2HPO4·12H2O、NaH2PO4·2H2O, boric acid, boron
Sand, tween 20, Triton X-100, sucrose, ultra-pure water；
Consumptive material: nitrocellulose filter (NC film), 8975 glass fibre, GF-06 glass fibre, adsorptive pads, base plate, capacity
Bottle, conical flask, EP pipe, rifle head.
2, the preparation before preparation
Prepared by 2.1 gold colloidals
Weigh 0.105g citrate three sodium in 15mlEP pipe, add 5ml pure water, mix to being completely dissolved and be 2% lemon
Solution is received in lemon acid three.
Take 40ml pure water in 50ml volumetric flask, in volumetric flask, add 0.5ml 1% chlorauric acid solution, use pure water constant volume
To 50ml, mixing is 0.01% chlorauric acid solution.
50ml 0.01% chlorauric acid solution is poured in 100ml conical flask, is placed on electric furnace and is heated to boiling with 3-4 shelves,
Draw 380 μ L 2% citrate three sodium solution with liquid-transfering gun, be quickly driven in the chlorauric acid solution of boiling, continue heating, i.e.
40-55nm gold colloidal is prepared complete.
2.2PBS buffer:
Weigh Na2HPO4·12H2O 1.615-2g, NaH2PO4·2H2O 0.225-5g, NaCl 4.0-5.0g is in beaker
In, add the dissolving of 400ml ultra-pure water and be settled to 500ml, standby after 0.22 μm membrane filtration.
3, Optimal pH determines:
Take 1mL gold colloidal respectively in 6 1.5mLEP pipes, be separately added into 0.2M K to often pipe2CO3 1μL、2μL、3μL、5
μ L, 7 μ L, 10 μ L, corresponding pH respectively may be about 7.0,7.5,8.0,8.5,9.0,9.5, is separately added into 150-200 μ to often pipe
L0.1mg/mL antibody-solutions, mixing, 37 degree stand 20min, are separately added into 100-130 μ L10%NaCl solution to often pipe, mixed
Even, stand and observe color change, 0.2M K2CO3Addition be 1-3 μ L EP pipe in gold colloidal color still for aubergine, other pipes
Becoming blue, optimum pH is 7.0-8.0.
4, minimum antibody labeling amount determines:
Take 4 EP pipes, take 1mL gold colloidal respectively and add in each pipe, then be separately added into K to often pipe2CO32-4 μ L, mixing, add
Enter the antibody-solutions of the 0.08-0.1mg/ml of 30 μ L (3 μ g), 50 μ L (5 μ g), 70 μ L (7 μ g), 100 μ L (10 μ g), mixing, 37
DEG C stand 20min, to often pipe add 10%NaCl solution, mixing, stand observe color change, antibody addition is 7-10 μ g
In EP pipe, gold colloidal color does not occurs significant change to be aubergine, antibody addition be 3,5 μ g EP pipe in gold colloidal become indigo plant, say
The minimum antibody addition of bright 1ml gold colloidal is 5-8 μ g.
5, solution preparation
0.2M boric acid solution is prepared: weigh boric acid 12.37g in beaker, adds after 800ml ultra-pure water dissolves and is settled to
1L, standby after 0.22 μm membrane filtration.
0.05M borax soln is prepared: weigh Borax 19.07g in beaker, adds after 800ml ultra-pure water dissolves and is settled to
1L, standby after 0.22 μm membrane filtration.
10mM pH8.0 boric acid-borate buffer solution preparation: take 3ml 0.05M borax soln, 7ml 0.2M boric acid respectively molten
Liquid, in beaker, adds 190ml ultra-pure water, mixes standby.
10mM pH8.4 boric acid-borate buffer solution preparation: take 4.5ml 0.05M borax soln, 5.5ml0.2M boric acid respectively
Solution, in beaker, adds 190ml ultra-pure water, mixes standby.
6, preparation method
1., the preparation of sample pad 2:
Preparation sample liquid: weigh 0.9g ± 0.05gNaCl, 0.2g ± 0.05 urea peroxide respectively in 100ml beaker, add
Enter 70ml 10mM pH8.4 borate buffer solution to dissolve, be separately added into 5ml horse serum, 1ml tween 20 and 1ml Triton X-
100, mixing, it is settled to 100ml, standby after 0.22 μm membrane filtration；
Again the glass fibre membrane after being cut into 0.7cm and 3.2cm width strip is placed in sample liquid, soaking at room temperature 4h,
37 DEG C of drying, save backup in drying basin.
2., the preparation of pad 3:
Step one, preparation pad 3 treatment fluid: weigh 0.1g ± 0.05gBSA, 0.5g ± 0.05g trehalose in 100ml
In beaker, add 70ml 10mM pH8.0 borate buffer solution and dissolve, add 0.3ml tween 20, mixing, be settled to 100ml,
After 0.22 μm membrane filtration standby；
Step 2, the preparation of gold labeling antibody: take 1mL 40-55nm gold colloidal in EP pipe, add 2 μ L0.2M K2CO3Molten
Liquid, mixing, add 70 μ L 0.08-0.1mg/ml β-HCG antibody-solutions, mixing, 37 DEG C stand 20min, add 100 μ L 10%
BSA solution (final concentration 1%), mixing, place 1.5h for 37 DEG C, 4 DEG C, 12000g is centrifuged 20min, abandons supernatant, adds 1ml PBS multiple
Molten, 4 DEG C, 12000g is centrifuged 20min, abandons supernatant, adds 100 μ L redissolution liquid and redissolves, and 4 DEG C save backup；
Prepared by step 3, CNB Nanoparticle labeling antibody: take the most ultrasonic 0.1%CNB nano-particle, first takes the centrifugal of 5ml
Pipe, is sequentially added into the β-hCG antibody 500-600ul of 20-30mM, 9.0pH borate 600-800ul and 1mg/ml, finally distinguishes
Add standby ultrasonic nano material 300ul, mix homogeneously.Hatch 2h for 37 DEG C, take out mixing every 10 minutes for a moment.Hatch
Cheng Hou, adds 2% mice serum confining liquid (or 0.5%CNB granule confining liquid) in centrifuge tube, is 6-8ml.37 DEG C of closings
2h, took out mixing for a moment, closes and terminate rear centrifugal treating every 10 minutes.14000g is centrifuged 15-25min, removes supernatant as far as possible,
Precipitation 30-50mM, pH9-10.0 borate dissolves and recovers volume, mix homogeneously.Precipitation is with preserving liquid, borate buffering
Liquid, each 300ul of trehalose dissolves, is placed in 4 DEG C of preservations.
Pad 3 raw material is cut into strip wide for 0.7cm again and is placed on soaking at room temperature 4h in pad 32 treatment fluid, 37 DEG C
Dry, then with drawing film metal spraying machine by golden labeling antibody-CNB Nanoparticle labeling antibody+Avidin (1mg/ml/) according to the amount of 12 μ L/cm
It is sprayed on pad 3, after 37 degree of drying, drying basin saves backup.
3., nitrocellulose filter 4 is coated:
Step one, preparation are coated buffer: weigh 3g trehalose in beaker, add after 80-100mlPBS dissolves and add
3-5ml methanol, mixing, it is settled to 100ml, standby after 0.22 μm membrane filtration；
Prepared by step 2, biotin antibody: take а-HCG antibody about 100 μ L in EP pipe with liquid-transfering gun；Take out from-20 degree
Biotin solution is redissolved, and 20mg/ml takes 1 μ L biotin solution respectively and joins in а-HCG antibody-solutions, mixing (biotin: а-
HCG antibody ≈ 20:1), room temperature is placed 1h and is proceeded in bag filter, by 4 DEG C of dialysed overnight of PBS；From bag filter take out biotin-
Antibody, 4 DEG C save backup.
Step 3, it is coated detection line 41: be diluted to 0.8-with being coated the biotin antibody that step 2 prepared by buffer
1.3mg/ml, draws on described nitrocellulose filter 4 by the amount of 0.9-1.5 μ L/cm, preserves standby after 37 DEG C of drying in drying basin
With.It is coated nature controlling line 42: with being coated buffer, sheep anti mouse polyclonal antibody is diluted to 1.2-1.5mg/ml, by 0.7-1.4 μ L/
The amount of cm is drawn on described nitrocellulose filter 4, saves backup after 37 DEG C of drying in drying basin；
7, the preparation of reagent paper:
In temperature 20-30 DEG C, the humidity hothouse less than 30-40%, take base plate 1, the most coated described nitric acid is fine
Dimension element film 4 is placed on the middle part of base plate 1 and pastes；
Paste described sample suction pad 5 in described nitrocellulose filter 4 side, and described sample suction pad 5 side is fine with described nitric acid
Dimension element film 4 overlaps, and opposite side edge is concordant with described base plate 1, and described nitrocellulose filter 4 opposite side is taken with described sample pad 2
Connect so that sample to be tested passes sequentially through described sample pad 2, described nitrocellulose filter 4, described sample suction pad 5 form the first level
Flow channel；
Described barrier film 6 two ends are overlapped on above described sample pad 2, described nitrocellulose filter 4 respectively, described pad
3 two ends are overlapped on above described barrier film 6, described nitrocellulose filter 4 respectively, and during detection, buffer passes sequentially through described knot
Conjunction pad 3, described nitrocellulose filter 4, described sample suction pad 5 form the second bottom horizontal flow sheet passage, finally with cutter by preparation
Reagent paper cuts into the test strips that 4mm is wide, and the test strips cut can load in plastic clip or rod (pen), forms quick diagnosis test card
Or rod (pen).
8, reagent paper test and result judge
Test strips is put into liquid to be measured (making well 10 be immersed under sample liquid level) stops 5s, take out and lie in table top
On, when, after the color change of described sample suction pad 5, taking 50 μ LPBS buffer and drop on buffer liquid pad, after 5min, read result,
In 30min effectively.
Result judges:
When two red stripes occur, one be positioned at detection line 41, another be positioned at nature controlling line 42, show conceived；When
One red stripes occurs, is only located at nature controlling line 42, shows there is no pregnancy；At nature controlling line 42 redfree band when 5-10 minute
Occur, show that incorrect operating process or test strips are rotten and damage, need correctly to operate by new test strips and/or weight
New test.
Two, the sensitivity to people's saliva chorionic-gonadotropin hormone (HCG) reagent paper of above-mentioned preparation is tested
1, saliva gathers: puts in saliva dilution liquid after gathering saliva with saliva acquisition rod, is existed respectively on acquisition rod two sides
Rub 5-6 time equipped with on the tube wall of saliva dilution liquid.
2, mark-on saliva: taking 98 μ L salivas in micropore, the HCG standard substance adding 2.5 μ L 1IU/ml i.e. obtain mark-on
The saliva of 25mIU/ml；Take 98 μ L, 95 μ L salivas respectively in micropore, add 2 μ L, the HCG standard substance of 5 μ L 100m IU/ml
Obtain the saliva of mark-on 2mIU/ml, 5mIU/ml；
3, test: keep flat on desktop by dual pathways test strips, takes the sample pad 2 that 50 μ L salivas dropwise put at sample channel and pads
On, room temperature stands 1-5min, after sample suction pad 5 has color change, then takes 50 μ LPBS buffer and dropwise puts the buffering in reagent passage
On fluid cushion, room temperature stands 5min, and observed result, saliva sensitivity can reach 2mIU/ml.Retest 10 times (such as table 1).
Table 1 people's saliva chorionic-gonadotropin hormone test strips sensitivity test
Note: " +++ " represents it is apparent that " ++ " represents visible, "+" represent faint visible, " " represents invisible.(table 1
In, C line represents that nature controlling line 42, T line represent detection line 41)
Three, 37 DEG C of stability of people's saliva chorionic-gonadotropin hormone (HCG) reagent paper of above-mentioned preparation are tested
Sample diluting liquid is 10mM, pH7.4PBS, 1%BSA, 0miu/ml hCG standard substance, 25miu/ml hCG standard
Product.6 test strips are taken from exsiccator, standby.Prepare 6 parts of detection sample (0miu/mlhCG standard substance；25miu/ml hCG marks
Quasi-product), test strips test 15min.Test result is as shown in table 2:
Table 2 people's 37 DEG C of stability tests of saliva chorionic-gonadotropin hormone test strips
Note: " +++ " represents clearly visible.(in table 2, C line represents that nature controlling line 42, T line represent detection line 41)
Result indicating signal is normal, does not has false positive, test strips to have good stability.
Four, the specificity of people's saliva chorionic-gonadotropin hormone (HCG) reagent paper of above-mentioned preparation is tested
Choose following reagent respectively:
HCG, A liquid: 0miu/ml hCG, 10mM PBS, pH7.4,1%BSA；HCG, B liquid: 25miu/ml hCG, 10mM
PBS, pH7.4,1%BSA；
HLH, A liquid: 0miu/ml hCG, 500miu/ml LH, 10mM PBS, pH7.4,1%BSA；HLH, B liquid: 25miu/
Ml hCG, 500miu/ml LH, 10mM PBS, pH7.4,1%BSA；
HFSH, A liquid: 0miu/ml hCG, 1000miu/ml FSH, 10mM PBS, pH7.4,1%BSA；HFSH, B liquid:
25miu/ml hCG, 1000miu/ml FSH, 10mM PBS, pH7.4,1%BSA；
HTSH, A liquid: 0miu/ml hCG, 1000uiu/ml TSH, 10mM PBS, pH7.4,1%BSA；HTSH, B liquid:
25miu/ml hCG, 1000uiu/ml TSH, 10mM PBS, pH7.4,1%BSA.
8 test strips are taken from exsiccator, standby.Prepare 4 groups of AB liquid detection samples, test strips test 15min (country's mark
Accurate: test A liquid T line does not develops the color；Test b liquid T line develops the color.), test result is as shown in table 3, table 4:
Table 3 people's saliva chorionic-gonadotropin hormone test strips feminine gender specificity is tested
Note: " +++ " represents it is apparent that " " represents invisible.
Table 4 people's saliva chorionic-gonadotropin hormone test strips positive specificity is tested
Note: " +++ " represents clearly visible.(in table 3,4, C line represents that nature controlling line 42, T line represent detection line 41)
Test result all meets Standard, and specificity is good, and test hCG with LH/FSH/TSH is equal for test strips the most of the present invention
Without intersecting.Result is consistent in triplicate.
Embodiment two: in double combination pad, signal during Avidin-Biotin system amplifies test
For reagent paper of the present invention uses the signal of pad (being divided into A pad and B pad in structure) amplify test, side
Method with reference to previous embodiment one, reagent source: 1. 100nm gold colloidal (Nano Composix, LN:lXW0093), 2. 1
R2CNB (cellulose nano-particle Asahi Kasei, LN130212-R2-1), 3. NHS-PE4-Biotin (Thermo-
Scientific, LN05192370), 4. streptaridin (Thermo-scientific, LNOE185707), 5. ESE reading
Instrument, model ESLF34-MB-4501.Test sample is the variable concentrations HCG standard substance with lXPBS, 1%BSA preparation.During test
Pad is divided into different combinations, such as following table:
Table 5A is the combination of different pads
It is following (table 5B, table 5C) that the signal of different pad combinations amplifies test result:
The signal amplification of the pad combination that table 5B systematic comparison is different
The abstract of table 5C average T line peak value
Conclusion: 1. group is the combination susceptiveness of A pad (CNB+ biotin antibody)+B pad (CNB+ Avidin)
The highest, and resoluting signal is best between HCG concentration 0-0.5MIU/ml.
Embodiment three: the clinical experiment of people saliva HCG
People's saliva chorionic-gonadotropin hormone (HCG) immune chromatography test paper of Application Example one preparation, carry out hospital and face
Bed detection test, uses this test strips to detect according to using method, with hospital clinical blood the clinical saliva sample collected
Fluid inspection result compares, and hospital's blood examination result shows that the blood-prostatae barrier value having 5 parts of samples in 50 parts of samples is less than 1.2mIU/ml,
Being diagnosed as not conceived, for negative sample, the blood-prostatae barrier value of other 45 parts of samples is more than 5mIU/ml, is diagnosed as pregnancy, for positive sample
This；The saliva of these 50 parts of samples is used respectively this ELISA test strip, 5 parts of negative sample of test strips detection and hospital clinical result
Unanimously and false positive do not occur, other 45 parts of samples of test strips detection are positive consistent with hospital clinical result and do not occur
False negative, HCG line coincidence rate 100%, test strips clinical trial see table 6.
Table 6 test strips clinical test results
Note: "-" represents invisible, " +++ " represents clearly visible, " ++ " expression is visible "+" represent faint visible.(table 6
In, C line represents that nature controlling line 42, T line represent detection line 41)
Embodiment four: the detection (detection of the 8th kind of form of one pad of preparation of people's saliva ELISA HBV
Reagent paper)
Mixing in healthy human saliva by the ELISA HBV of various variable concentrations to be detected, the present embodiment is not only
Successfully be detected ELISA HBV, and sensitivity is high, the least concentration wherein recorded is that hepatitis B surface antigen HbsAg is
1ng/ml；Surface antibody HbsAb is 10miu/ml；E antigen HbeAg is 1NCU/ml；E antibody HbeAb is 2NCU/ml；Core resists
Body HbcAb is 1NCU/ml.Meanwhile, comparing with enzyme linked immunosorbent assay (Elisa), detect above-mentioned sample, the two positive findings is such as
Following table:
| The present invention | Elisa | |
| HbsAg | 33 | 35 |
| HbeAg | 6 | 7 |
| HbsAb | 15 | 15 |
| HbeAb | 11 | 12 |
| HbcAb | 29 | 30 |
From upper table, the two testing result is essentially identical.
Described A pad (32), described B pad (31) or described pad (3), described nitrocellulose filter (4)
Middle application Avidin or biotin.
Above, tracer label thing selectivity application gold colloidal, cellulose nano-particle, latex particle or gold colloidal are fine
Dimension element nano-particle etc..
The foregoing is only presently preferred embodiments of the present invention, not in order to limit the present invention, all spirit in the present invention and
Within principle, any modification, equivalent substitution and improvement etc. made, should be included within the scope of the present invention.
Claims (10)
1. a hypersensitivity immunity chromatography detection test paper, it is characterised in that: base plate (1), and it is successively set on described base plate
(1) sample pad (2) on, pad (3), nitrocellulose filter (4), sample suction pad (5), described sample pad (2), described nitric acid are fine
Dimension element film (4), described sample suction pad (5) overlap successively, constitute one for the passage that liquid stream to be measured leads to, the i.e. first horizontal channel；
Wherein, also including that a barrier film (6), described barrier film (6) one end are overlapped in described sample pad (2), the other end is overlapped on institute
Stating on nitrocellulose filter (4), and described pad (3) one end is overlapped on described barrier film (6), the other end is overlapped on described nitre
On acid cellulose film (4), constitute another between described pad (3), described nitrocellulose filter (4), described sample suction pad (5) and lead to
Road, the i.e. second horizontal channel so that constitute dual channel mode Test paper.
A kind of hypersensitivity immunity chromatography detection test paper, it is characterised in that: described sample pad (2)
On be provided with a filter pad (7), for filtering the chaff interference in detected material.
A kind of hypersensitivity immunity chromatography detection test paper, it is characterised in that: on described barrier film (6)
Also being pasted with a buffer liquid pad (8), and described buffer liquid pad (8) one end is overlapped on described barrier film (6), the other end is overlapped on institute
State on pad (3).
A kind of hypersensitivity immunity chromatography detection test paper, it is characterised in that: on described barrier film (6)
Also being pasted with a buffer liquid pad (8), and described buffer liquid pad (8) one end is overlapped on described barrier film (6), the other end is overlapped on institute
State on pad (3).
5. according to hypersensitivity immunity chromatography detection test paper a kind of described in claim 1 or 2 or 3, it is characterised in that: described combination
Pad (3) also includes that B pad (31), A pad (32), described B pad (31) one end are overlapped on described nitrocellulose filter
(4), on, described B pad (31) other end is overlapped by one end of described A pad (32), and described A pad (32) is another
One end is overlapped on described barrier film (6), thus constitutes double combination pad.
A kind of hypersensitivity immunity chromatography detection test paper, it is characterised in that: described pad (3)
Also include that B pad (31), A pad (32), described B pad (31) one end are overlapped on described nitrocellulose filter (4),
Described B pad (31) other end is overlapped by one end of described A pad (32), and the other end of described A pad (32) is taken
It is connected on described barrier film (6), thus constitutes double combination pad.
A kind of hypersensitivity immunity chromatography detection test paper, it is characterised in that: described Test paper is also
Including a shell (9), and described shell (9) include offering successively well (10), liquid filling hole (11), detection form (12),
Sample suction pad form (13), and described well (10) is corresponding with described sample pad (2), described detection form (12) and described nitric acid
The upper coated described detection line (41) of cellulose membrane (4) is corresponding with described nature controlling line (42), for by described detection form
(12) testing result is observed；
Described sample suction pad form (13) is corresponding with described sample suction pad (5), is used for observing whether sample reaches described sample suction pad (5).
A kind of hypersensitivity immunity chromatography detection test paper, it is characterised in that: when setting on Test paper
When being equipped with described filter pad (7), described well (10) is corresponding with described filter pad (7) so that sample passes through described well
(10) carry out chromatographing or pass sequentially through described filter pad (7) after, described sample pad chromatographs after (2).
9. according to hypersensitivity immunity chromatography detection test paper a kind of described in claim 7 or 8, it is characterised in that: described liquid filling hole
(11) corresponding with described pad (3), when described pad (3) is divided into A pad (32), B pad (31), described liquid feeding
Hole (11) is corresponding with described A pad (32), when being provided with buffer liquid pad (8) on described pad (3), and described liquid filling hole
(11) corresponding with described buffer liquid pad (8), for buffer by described buffer liquid pad (8) enter described pad (3) or
Chromatograph after A pad (32) is upper.
A kind of hypersensitivity immunity chromatography detection test paper, it is characterised in that:
Tracer label thing on described pad (3), described nitrocellulose filter (4) be gold colloidal or cellulose nano-particle or
Latex particle or gold colloidal cellulose nano-particle；Described A pad (32), described B pad (31) or described combination
Application Avidin or biotin in pad (3), described nitrocellulose filter (4).
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