CN103278637B - A kind of carbon nano-tube Test paper detecting helicobacter pylori and preparation method thereof - Google Patents
A kind of carbon nano-tube Test paper detecting helicobacter pylori and preparation method thereof Download PDFInfo
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- CN103278637B CN103278637B CN201310221645.3A CN201310221645A CN103278637B CN 103278637 B CN103278637 B CN 103278637B CN 201310221645 A CN201310221645 A CN 201310221645A CN 103278637 B CN103278637 B CN 103278637B
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 104
- 239000002041 carbon nanotube Substances 0.000 title claims abstract description 91
- 229910021393 carbon nanotube Inorganic materials 0.000 title claims abstract description 91
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 241000590002 Helicobacter pylori Species 0.000 title claims abstract description 15
- 229940037467 helicobacter pylori Drugs 0.000 title claims abstract description 15
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 47
- 239000000020 Nitrocellulose Substances 0.000 claims abstract description 38
- 229920001220 nitrocellulos Polymers 0.000 claims abstract description 38
- 239000002238 carbon nanotube film Substances 0.000 claims abstract description 24
- 206010019375 Helicobacter infections Diseases 0.000 claims abstract description 20
- 238000001514 detection method Methods 0.000 claims abstract description 14
- 239000002131 composite material Substances 0.000 claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 11
- 238000000926 separation method Methods 0.000 claims abstract description 7
- 239000000243 solution Substances 0.000 claims description 22
- 229910052799 carbon Inorganic materials 0.000 claims description 13
- 238000001035 drying Methods 0.000 claims description 11
- 239000012528 membrane Substances 0.000 claims description 10
- 239000012467 final product Substances 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 7
- 239000000725 suspension Substances 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 239000004342 Benzoyl peroxide Substances 0.000 claims description 6
- OMPJBNCRMGITSC-UHFFFAOYSA-N Benzoylperoxide Chemical compound C=1C=CC=CC=1C(=O)OOC(=O)C1=CC=CC=C1 OMPJBNCRMGITSC-UHFFFAOYSA-N 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 235000019400 benzoyl peroxide Nutrition 0.000 claims description 6
- 238000013016 damping Methods 0.000 claims description 6
- 239000000706 filtrate Substances 0.000 claims description 6
- 239000012530 fluid Substances 0.000 claims description 6
- 239000002048 multi walled nanotube Substances 0.000 claims description 6
- 239000007921 spray Substances 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 239000006185 dispersion Substances 0.000 claims description 5
- 239000007800 oxidant agent Substances 0.000 claims description 5
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 3
- 206010013786 Dry skin Diseases 0.000 claims description 3
- 229920001213 Polysorbate 20 Polymers 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 229940098773 bovine serum albumin Drugs 0.000 claims description 3
- 229920002301 cellulose acetate Polymers 0.000 claims description 3
- 239000000084 colloidal system Substances 0.000 claims description 3
- 238000004090 dissolution Methods 0.000 claims description 3
- 239000003365 glass fiber Substances 0.000 claims description 3
- 239000010931 gold Substances 0.000 claims description 3
- 229910052737 gold Inorganic materials 0.000 claims description 3
- 230000033444 hydroxylation Effects 0.000 claims description 3
- 238000005805 hydroxylation reaction Methods 0.000 claims description 3
- 230000007935 neutral effect Effects 0.000 claims description 3
- 230000010355 oscillation Effects 0.000 claims description 3
- 239000008363 phosphate buffer Substances 0.000 claims description 3
- 239000004033 plastic Substances 0.000 claims description 3
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 3
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 3
- 238000001556 precipitation Methods 0.000 claims description 3
- 239000002109 single walled nanotube Substances 0.000 claims description 3
- 150000002148 esters Chemical class 0.000 claims description 2
- 239000000463 material Substances 0.000 abstract description 7
- 230000035945 sensitivity Effects 0.000 abstract description 7
- 102000004169 proteins and genes Human genes 0.000 abstract description 5
- 108090000623 proteins and genes Proteins 0.000 abstract description 5
- 230000000274 adsorptive effect Effects 0.000 abstract description 3
- 238000003317 immunochromatography Methods 0.000 abstract description 3
- 238000002372 labelling Methods 0.000 abstract description 3
- 244000052616 bacterial pathogen Species 0.000 abstract description 2
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 2
- 229920002678 cellulose Polymers 0.000 description 3
- 239000001913 cellulose Substances 0.000 description 3
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 description 2
- 235000010413 sodium alginate Nutrition 0.000 description 2
- 239000000661 sodium alginate Substances 0.000 description 2
- 229940005550 sodium alginate Drugs 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 238000002604 ultrasonography Methods 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000007306 functionalization reaction Methods 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 239000004816 latex Substances 0.000 description 1
- 229920000126 latex Polymers 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000000700 radioactive tracer Substances 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
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Abstract
The present invention relates to a kind of carbon nano-tube Test paper detecting helicobacter pylori and preparation method thereof, specifically, relate to a kind of carbon nano-tube method detection reagent material detecting spiral helicobacter pylori infection for immunochromatography technique.It is characterized in that: this test paper base plate be pasted with successively sample pad, carbon nano-tube colloidal gold pad, carbon nano-tube film, nitrocellulose filter and inhale sample pad; Described carbon nano-tube colloidal gold pad is attached with the spiral helicobacter pylori infection monoclonal antibody 1 of carbon nano-tube colloidal gold composite mark, described carbon nano-tube film is adsorbed the filter paper of carbon nano-tube, described nitrocellulose filter is coated with detection line and the nature controlling line of spiral helicobacter pylori infection monoclonal antibody 2.The object of the present invention is to provide a kind of carbon nano-tube colloidal gold composite, as trace labelling material, for marking pylori spiral bacilli antibody, then through carbon nano-tube film, the performance of adsorptive separation protein can be improved, improve the sensitivity detected.
Description
Technical field
The present invention relates to a kind of carbon nano-tube Test paper detecting helicobacter pylori and preparation method thereof, specifically, relate to a kind of carbon nano-tube method detection reagent material detecting spiral helicobacter pylori infection for immunochromatography technique.
Background technology
Immunochromatography technique is widely used in multiple fields such as disease markers detection, food inspection, scientific research, selected trace labelling material is generally collaurum, latex particle etc., chromatographic material is generally cellulose membrane, comprise nitrocellulose filter, cellulose acetate membrane, cellulose mixture film etc., wherein the most frequently used is nitrocellulose filter.Under current technical conditions, the tracer materials such as collaurum and nitrocellulose filter is used to be difficult to improve detection sensitivity again as carrier.
Summary of the invention
Technical matters to be solved by this invention is: provide a kind of carbon nano-tube colloidal gold composite, as trace labelling material, for marking pylori spiral bacilli antibody, then through carbon nano-tube film, the performance of adsorptive separation protein can be improved, improve the sensitivity detected.
For achieving the above object, the present invention is by the following technical solutions:
Detect a carbon nano-tube Test paper for helicobacter pylori, it is characterized in that: this test paper base plate be pasted with successively sample pad, carbon nano-tube colloidal gold pad, carbon nano-tube film, nitrocellulose filter and inhale sample pad; Described carbon nano-tube colloidal gold pad is attached with the spiral helicobacter pylori infection monoclonal antibody 1 of carbon nano-tube colloidal gold composite mark, described carbon nano-tube film is adsorbed the filter paper of carbon nano-tube, described nitrocellulose filter is coated with detection line and the nature controlling line of spiral helicobacter pylori infection monoclonal antibody 2.
Carbon nano-tube used is the one in Single Walled Carbon Nanotube, multi-walled carbon nano-tubes, hydroxylation carbon nano-tube, carboxylic carbon nano-tube, water-soluble carbon nanometer tube.
Carbon nano-tube colloidal gold composite is that carbon nano-tube and collaurum form carbon nano-tube colloidal gold composite by assembling.
Described carbon nano-tube film filter paper film adsorbing carbon nanotubes suspension, both obtains after drying.
Water-soluble carbon nanometer tube is the carbon nano-tube after strong oxidizer process, and strong oxidizer used is the one in sulfonitric, benzoyl peroxide.
Filter paper film used is the one in nitrocellulose filter, cellulose acetate membrane, mixed ester film, ordinary filter paper.
A preparation method for the carbon nano-tube Test paper of detection helicobacter pylori described above, is characterized in that: on test paper base plate, paste sample pad, carbon nano-tube colloidal gold pad, carbon nano-tube film, nitrocellulose filter successively and inhale sample pad; Then prepare respectively:
(1), the preparation of water-soluble carbon nanometer tube:
Take 0.3 gram of benzoyl peroxide in the Erlenmeyer flask of 50ml, and add 10ml absolute ethyl alcohol, supersonic oscillations are to dissolving completely, the multi-walled carbon nano-tubes of 0.003 gram is added wherein, after 80 DEG C of backflow 5 h, is cooled to 4 DEG C, filters, rinse with absolute ethyl alcohol and reach neutral to filtrate, collect filtrate, be settled to 50ml with water, obtain final product;
(2), the preparation of carbon nano-tube colloidal gold composite:
Get the 40nm colloidal gold solution 50ml that mass percent is 0.01%, add in above-mentioned water-soluble carbon nanometer tube solution, stir 10 minutes, centrifugal 30 minutes of 12000rpm, separation of supernatant, precipitation pH value be 7.4 PBS damping fluid redissolve to 50ml, to obtain final product;
(3), the preparation of carbon nano-tube film:
The shortwall carbon nano-tube of 100mg is added 50ml, 0.5% Tween-20 aqueous solution in, within ultrasonic 5 minutes, make its abundant dispersion suspension, be immersed by ordinary filter paper, take out, rinse out non-adsorbing carbon nanotubes with water, dry, slitting is for subsequent use;
(4), the preparation of carbon nano-tube colloid gold label spiral helicobacter pylori infection monoclonal antibody 1:
Get 50ml carbon nano-tube colloidal gold solution in beaker, with the K of 0.1mol/L
2cO
3solution is adjusted to pH8.2, adds 0.5mg spiral helicobacter pylori infection monoclonal antibody in solution
1, stirring at room temperature 30 minutes, add percentage by weight be 5% bovine serum albumin(BSA) (BSA) close 1 hour, 12000rpm, 4 DEG C centrifugal 30 minutes, abandon supernatant, with damping fluid redissolve to 50ml, by 1ml solution paving 30cm
2ratio, be layered on glass fibre membrane by dissolution homogeneity, 37 DEG C of dryings 30 minutes, make carbon nano-tube colloidal gold pad;
(5), the bag quilt of nitrocellulose filter:
The spiral helicobacter pylori infection antibody 2 matched with antibody 1 is dissolved in the solution that 0.01M, pH8.2 phosphate buffer is mixed with 0.1mg/ml, rule with the parameter of 1 μ l/cm at nitrocellulose filter with spray film instrument, for detection line, simultaneously on nitrocellulose membrane, spray is drawn two anti-as nature controlling line, after line by nitrocellulose filter drying at room temperature 8 hours;
(6), the assembling of Test paper:
In hothouse, temperature 20 ~ 25 DEG C, humidity is less than 30%, sample pad, carbon nano-tube colloidal gold pad, carbon nano-tube film, nitrocellulose filter and thieving paper are pasted onto on base plate successively, wherein nitrocellulose filter is positioned in the middle part of base plate, nitrocellulose filter is coated with one end of T line and 1/3 of carbon nano-tube colloidal gold pad overlaps, the one end being coated with C line overlaps with inhale sample pad 1/10, carbon nano-tube film is placed on below carbon nano-tube colloidal gold pad, and extending to 4mm on nitrocellulose filter, 1/5 of sample pad and colloidal gold pad overlaps; Finally be cut into the test strips that width is 4mm, also test strips can be loaded in a plastic clip and make test card.
The carbon nano-tube of the application is a kind of novel nano-material, lightweight, and hexagonal structure connects perfect, has many abnormal mechanics, electricity and chemical property.Carbon nano-tube is existing many research in immunoassay, mainly concentrates on immunosensor field, can increase substantially detection sensitivity.Carbon nano-tube is divided into Single Walled Carbon Nanotube, multi-walled carbon nano-tubes, hydroxylation carbon nano-tube, carboxylic carbon nano-tube, also can obtain water-soluble carbon nanometer tube after treatment, has good adsorption separation performance, and the carbon nano-tube of functionalization more easily and protein bound.The compound labelled protein of carbon nano-tube and collaurum, then through carbon nano-tube film, the performance of adsorptive separation protein can be improved, improve the sensitivity detected.
embodiment:
A kind of carbon nano-tube Test paper detecting helicobacter pylori and preparation method thereof, this test paper base plate is pasted with successively sample pad, carbon nano-tube colloidal gold pad, carbon nano-tube film, nitrocellulose filter and inhales sample pad; Described carbon nano-tube colloidal gold pad is attached with the spiral helicobacter pylori infection monoclonal antibody 1 of carbon nano-tube colloidal gold composite mark, described carbon nano-tube film is adsorbed the filter paper of carbon nano-tube, described nitrocellulose filter is coated with detection line and the nature controlling line of spiral helicobacter pylori infection monoclonal antibody 2.
Method 1: by mass percent be 0.1 ~ 99.9% carbon nano-tube surfactant or shitosan, sodium alginate ultrasound wave make its abundant dispersion suspension, by mass percent be 0.1 ~ 99.9% collaurum add wherein, stir homogenate, centrifuging, is drying to obtain.
Method 2: by mass percent be 0.1 ~ 99.9% carbon nano-tube surfactant or shitosan, sodium alginate ultrasound wave make its abundant dispersion suspension, filter paper film is immersed, adsorbing carbon nanotubes, takes out, with water, the carbon nano-tube of not adsorbing is washed away, be drying to obtain.
Method 3: by carbon nano-tube strong oxidizer sulfonitric or benzoyl peroxide process, obtain water-soluble carbon nanometer tube, then cellulose membrane is placed in water-soluble carbon nanometer tube and soaks, dry after taking out, to obtain final product.
Embodiment;
the preparation of water-soluble carbon nanometer tube:
Take 0.3 gram of benzoyl peroxide in the Erlenmeyer flask of 50ml, and add 10ml absolute ethyl alcohol, supersonic oscillations are to dissolving completely, the multi-walled carbon nano-tubes of 0.003 gram is added wherein, after 80 DEG C of backflow 5 h, be cooled to 4 DEG C, filter, rinse with absolute ethyl alcohol and reach neutral to filtrate. collect filtrate, be settled to 50ml with water, obtain final product.
the preparation of carbon nano-tube colloidal gold composite:
Get the 40nm colloidal gold solution 50ml that mass percent is 0.01%, add in above-mentioned water-soluble carbon nanometer tube solution, stir 10 minutes, centrifugal 30 minutes of 12000rpm, separation of supernatant, precipitation pH value be 7.4 PBS damping fluid redissolve to 50ml, to obtain final product.
the preparation of carbon nano-tube film:
The shortwall carbon nano-tube of 100mg is added 50ml, 0.5% Tween-20 aqueous solution in, within ultrasonic 5 minutes, make its abundant dispersion suspension, ordinary filter paper be immersed, take out, rinse out non-adsorbing carbon nanotubes with water, be drying to obtain.Slitting is for subsequent use.
carbon nano-tube colloid gold label spiral helicobacter pylori infection monoclonal antibody 1
Get 50ml carbon nano-tube colloidal gold solution in beaker, with the K of 0.1mol/L
2cO
3solution is adjusted to pH8.2, adds 0.5mg spiral helicobacter pylori infection monoclonal antibody in solution
1, stirring at room temperature 30 minutes, add percentage by weight be 5% bovine serum albumin(BSA) (BSA) close 1 hour, 12000rpm, 4 DEG C centrifugal 30 minutes, abandon supernatant, with damping fluid redissolve to 50ml, by 1ml solution paving 30cm
2ratio, be layered on glass fibre membrane by dissolution homogeneity, 37 DEG C of dryings 30 minutes, make carbon nano-tube colloidal gold pad.
the bag quilt of nitrocellulose filter:
The spiral helicobacter pylori infection antibody 2 matched with antibody 1 is dissolved in the solution that 0.01M, pH8.2 phosphate buffer (PBS) is mixed with 0.1mg/ml, rule with the parameter of 1 μ l/cm at nitrocellulose filter with spray film instrument, for detection line (T line), draw two anti-as nature controlling line (C line) in far-end spray simultaneously, after line by nitrocellulose filter drying at room temperature 8 hours.
the assembling of Test paper:
In hothouse, temperature 20 ~ 25 DEG C, humidity is less than 30%, sample pad, carbon nano-tube colloidal gold pad, carbon nano-tube film, nitrocellulose filter and thieving paper are pasted onto on base plate successively, wherein nitrocellulose filter is positioned in the middle part of base plate, nitrocellulose filter is coated with one end of T line and 1/3 of carbon nano-tube colloidal gold pad overlaps, the one end being coated with C line overlaps with inhale sample pad 1/10, carbon nano-tube film is placed on below carbon nano-tube colloidal gold pad, and extending to 4mm on nitrocellulose filter, 1/5 of sample pad and colloidal gold pad overlaps.Finally be cut into the test strips that width is 4mm, also test strips can be loaded in a plastic clip and make test card.
Sensitivity test
The spiral helicobacter pylori infection antigen of preparation variable concentrations, detect with this Test paper, result shows that the sensitivity of this Test paper is 0.5ng/ml.Can be used for the helicobacter pylori detection that saliva makees sample.
Claims (7)
1. detect a carbon nano-tube Test paper for helicobacter pylori, it is characterized in that: this test paper base plate be pasted with successively sample pad, carbon nano-tube colloidal gold pad, carbon nano-tube film, nitrocellulose filter and inhale sample pad; Described carbon nano-tube colloidal gold pad is attached with the spiral helicobacter pylori infection monoclonal antibody 1 of carbon nano-tube colloidal gold composite mark, described carbon nano-tube film is adsorbed the filter paper of carbon nano-tube, described nitrocellulose filter is coated with detection line and the nature controlling line of spiral helicobacter pylori infection monoclonal antibody 2.
2. a kind of carbon nano-tube Test paper detecting helicobacter pylori according to claim 1, is characterized in that: carbon nano-tube used is the one in Single Walled Carbon Nanotube, multi-walled carbon nano-tubes, hydroxylation carbon nano-tube, carboxylic carbon nano-tube, water-soluble carbon nanometer tube.
3. a kind of carbon nano-tube Test paper detecting helicobacter pylori according to claim 1, is characterized in that: carbon nano-tube colloidal gold composite is that carbon nano-tube and collaurum form carbon nano-tube colloidal gold composite by assembling.
4. a kind of carbon nano-tube Test paper detecting helicobacter pylori according to claim 1, is characterized in that: described carbon nano-tube film filter paper film adsorbing carbon nanotubes suspension, after drying and get final product.
5. a kind of carbon nano-tube Test paper detecting helicobacter pylori according to claim 2, it is characterized in that: water-soluble carbon nanometer tube is the carbon nano-tube after strong oxidizer process, strong oxidizer used is the one in sulfonitric, benzoyl peroxide.
6. a kind of carbon nano-tube Test paper detecting helicobacter pylori according to claim 4, is characterized in that: filter paper film used is the one in nitrocellulose filter, cellulose acetate membrane, mixed ester film, ordinary filter paper.
7. detect a preparation method for the carbon nano-tube Test paper of helicobacter pylori as claimed in claim 1, it is characterized in that: on test paper base plate, paste sample pad, carbon nano-tube colloidal gold pad, carbon nano-tube film, nitrocellulose filter successively and inhale sample pad; Then prepare respectively:
(1), the preparation of water-soluble carbon nanometer tube:
Take 0.3 gram of benzoyl peroxide in the Erlenmeyer flask of 50ml, and add 10ml absolute ethyl alcohol, supersonic oscillations are to dissolving completely, the multi-walled carbon nano-tubes of 0.003 gram is added wherein, after 80 DEG C of backflow 5 h, is cooled to 4 DEG C, filters, rinse with absolute ethyl alcohol and reach neutral to filtrate, collect filtrate, be settled to 50ml with water, obtain final product;
(2), the preparation of carbon nano-tube colloidal gold composite:
Get the 40nm colloidal gold solution 50ml that mass percent is 0.01%, add in above-mentioned water-soluble carbon nanometer tube solution, stir 10 minutes, centrifugal 30 minutes of 12000rpm, separation of supernatant, precipitation pH value be 7.4 PBS damping fluid redissolve to 50ml, to obtain final product;
(3), the preparation of carbon nano-tube film:
The shortwall carbon nano-tube of 100mg is added 50ml, 0.5% Tween-20 aqueous solution in, within ultrasonic 5 minutes, make its abundant dispersion suspension, be immersed by ordinary filter paper, take out, rinse out non-adsorbing carbon nanotubes with water, dry, slitting is for subsequent use;
(4), the preparation of carbon nano-tube colloid gold label spiral helicobacter pylori infection monoclonal antibody 1:
Get 50ml carbon nano-tube colloidal gold solution in beaker, with the K of 0.1mol/L
2cO
3solution is adjusted to pH8.2, adds 0.5mg spiral helicobacter pylori infection monoclonal antibody in solution
1, stirring at room temperature 30 minutes, add percentage by weight be 5% bovine serum albumin(BSA) (BSA) close 1 hour, 12000rpm, 4 DEG C centrifugal 30 minutes, abandon supernatant, with damping fluid redissolve to 50ml, by 1ml solution paving 30cm
2ratio, be layered on glass fibre membrane by dissolution homogeneity, 37 DEG C of dryings 30 minutes, make carbon nano-tube colloidal gold pad;
(5), the bag quilt of nitrocellulose filter:
The spiral helicobacter pylori infection antibody 2 matched with antibody 1 is dissolved in the solution that 0.01M, pH8.2 phosphate buffer is mixed with 0.1mg/ml, rule with the parameter of 1 μ l/cm at nitrocellulose filter with spray film instrument, for detection line, simultaneously on nitrocellulose membrane, spray is drawn two anti-as nature controlling line, after line by nitrocellulose filter drying at room temperature 8 hours;
(6), the assembling of Test paper:
In hothouse, temperature 20 ~ 25 DEG C, humidity is less than 30%, sample pad, carbon nano-tube colloidal gold pad, carbon nano-tube film, nitrocellulose filter and thieving paper are pasted onto on base plate successively, wherein nitrocellulose filter is positioned in the middle part of base plate, nitrocellulose filter is coated with one end of T line and 1/3 of carbon nano-tube colloidal gold pad overlaps, the one end being coated with C line overlaps with inhale sample pad 1/10, carbon nano-tube film is placed on below carbon nano-tube colloidal gold pad, and extending to 4mm on nitrocellulose filter, 1/5 of sample pad and colloidal gold pad overlaps; Finally be cut into the test strips that width is 4mm, also test strips loaded in a plastic clip and make test card.
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