CN1746675A - Immune chromatograph testing strip and production thereof - Google Patents
Immune chromatograph testing strip and production thereof Download PDFInfo
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- CN1746675A CN1746675A CN 200410074522 CN200410074522A CN1746675A CN 1746675 A CN1746675 A CN 1746675A CN 200410074522 CN200410074522 CN 200410074522 CN 200410074522 A CN200410074522 A CN 200410074522A CN 1746675 A CN1746675 A CN 1746675A
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Abstract
An immune chromatographic test bar consists of supporter, drinking fibrous membrance, nitrate fibrous membrance, sample membrance, waterproof membrance and colloidal gold labeled membrance, Its preparing method utilizes colloidal gold labeling technique and quantitative immune competition method to produce test bar for being used in quick detection.
Description
Technical field
The present invention relates to a kind of immune chromatograph testing strip and manufacture method thereof, particularly having related to can fast detecting progestational hormone, test-strips and the manufacture method thereof of estrogen and micromolecule hormone.
Background technology
At present, people's living standard improves gradually, health care consciousness is strengthened gradually, a lot of nutriment and tonic accumulate educates and gives birth to, making people's health strengthen the life-span day by day generally improves, but also bring many negative interactions simultaneously, the estrogen taken of climacteric women for example, if dose does not have effect for a short time,, and detect female hormone now or the micromolecule hormone need pass through very complicated medical science testing process if the dose conference causes the canceration of system of gynaecology, and can not detect at any time, vast women blindly replenishes female hormone under situation about not considering the consequences, can make health be subjected to bigger injury.
Summary of the invention
Goal of the invention of the present invention provides a kind of immune chromatograph testing strip and manufacture method thereof that can make things convenient for, detect at any time the micromolecule hormone.
In order to realize this goal of the invention, the invention provides a kind of immune chromatograph testing strip, it comprises stilt, the water-absorption fiber film, nitrocellulose membrane, sample film, waterproof membrane and colloid gold label film, wherein: nitrocellulose membrane sticks to the stage casing of bar shaped stilt, the water-absorption fiber film sticks to the top of stilt, the lower end of water-absorption fiber film and the upper end of nitrocellulose membrane overlap joint, the colloid gold label film, waterproof membrane and sample film stick to end from stilt successively to stilt the nitrocellulose membrane, and an end of colloid gold label film and waterproof membrane overlap joint, one end of waterproof membrane and sample film overlap joint, one end of sample film and nitrocellulose membrane overlap joint, wherein the lap of splice of colloid gold label film one end and waterproof membrane sticks to length on the stilt greater than waterproof membrane, at least comprise a kind of pairing antibody of antigen with being detected of concentration and the antibody of a kind of anti-immune protein IgG on the described nitrocellulose membrane, comprise with colloid gold label on the colloid gold label film, the antigen that is detected and with the immune protein IgG of colloid gold label.
The invention provides a kind of immune chromatograph testing strip, wherein: the pairing antibody of the antigen with being detected that comprises three kinds of variable concentrations on the described nitrocellulose membrane.
The invention provides a kind of immune chromatograph testing strip, wherein: the pairing antibody of the antigen with being detected of described three kinds of concentration and the antibody of a kind of anti-immune protein IgG longitudinally evenly distribute to the absorbing membrane end from being positioned at the sample film end, the spraying of every kind of antibody highly is the 1-3 millimeter, is spaced apart the 2-3 millimeter between four kinds of antibody.
The invention provides a kind of immune chromatograph testing strip, wherein: described sample film and colloid gold label film are glass fibre membrane.
The invention provides a kind of immune chromatograph testing strip, wherein: described detection antigen is micromolecule antigen.
The invention provides a kind of immune chromatograph testing strip, wherein: described little antigen molecule antigen is progestational hormone antigen or estrogen antigen.
The invention provides a kind of method of making immune chromatograph testing strip, wherein: it comprises nitrocellulose membrane is sticked on the stage casing of bar shaped stilt, the water-absorption fiber film is sticked to the top of stilt, the lower end of water-absorption fiber film and the upper end of nitrocellulose membrane overlap joint, with the colloid gold label film, waterproof membrane and sample film stick to end from stilt successively to the stilt the nitrocellulose membrane, and an end of colloid gold label film and waterproof membrane overlap joint, one end of waterproof membrane and sample film overlap joint, one end of sample film and nitrocellulose membrane overlap joint, wherein the lap of splice of colloid gold label film one end and waterproof membrane sticks to length on the stilt greater than waterproof membrane, at least apply a kind of pairing antibody of antigen with being detected and a kind of anti-immune protein antibody of concentration on the described nitrocellulose membrane, the colloid gold label film is to apply on glass fibre membrane with the antigenic solution that is detected of colloid gold label with the immune protein solution of colloid gold label.
The invention provides a kind of method of making immune chromatograph testing strip, wherein: on described nitrocellulose membrane different parts, spray three kinds of various dose respectively with the antigen reactive antibody of colloid gold label and a kind of anti-immune protein antibody, described different parts is that every kind of antibody is from becoming to be wire near the sample film end to spraying successively near water-absorption fiber film end, three kinds of various dose be followed successively by detection line 1 with the antigen reactive antibody of colloid gold label, 2,3, a kind of anti-immune protein antibody is control line, four kinds of dosage of antibody are respectively the 0.3-0.6 microgram, 1.0-1.7 microgram, 2.6-3.9 microgram and 4.4-5.6 microgram, highly about 1 mm wide of the spraying of every line 3 millimeters long, control line is positioned at apart from nitrocellulose filter and water-absorption fiber film junction 3-6 millimeter, each line is 2～3 mm distance at interval, spraying nitrocellulose filter later is after drying, albumin solution with 40 grams per liters-60 grams per liter soaks sealing 1-3 hour, physiological solution with the 0.008-0.013 mole washs 2-4 time again, and the sealing of dry back is preserved.
The invention provides a kind of method of making immune chromatograph testing strip, wherein: be added on the glass fibre membrane after the immune protein solution with colloid gold label with the antigenic solution of colloid gold label and 20-60 microlitre of 20-60 microlitre mixed, in room temperature or temperature is 3-6 ℃ of drying down, makes the colloid gold label film.
The invention provides a kind of method of making immune chromatograph testing strip, wherein: the manufacturing step of the antigenic solution of described usefulness colloid gold label is as follows:
1. with micro-0.08-012mol/L acetate solution colloidal gold solution being adjusted to potential of hydrogen earlier is 8.2-8.6;
2. in 80-120 milliliter colloidal gold solution, add then in the protein solution that will detect antigen of 300-500ug, at room temperature stirred 10-15 minute;
3. the 0.8-1.3% polyglycol solution that adds the 4-10 milliliter again;
4. restrain centrifugal actions 20-40 minute through 10000～100000, the careful suction removed supernatant;
5. sediment is mixed with the damping fluid of 0.2～0.5mg/ml polyglycol of 80-120 milliliter again;
6. repeat with 10000～100000 gram centrifugal actions 20-40 minute, the careful suction removed supernatant;
7. after sediment being mixed with the damping fluid of 0.2～0.5mg/ml polyglycol of 80-120 milliliter, add the Sodium azide of 30-70 milligram again, reached anticorrosion purpose after the mixing, putting temperature is 3-6 ℃ of preservation.
The invention provides a kind of method of making immune chromatograph testing strip, wherein: the manufacturing step of the immune protein solution of described usefulness colloid gold label is as follows:
1. with micro-0.08-012mol/L acetate solution colloidal gold solution is adjusted to potential of hydrogen 7.8-8.2 earlier;
2. in 80-120 milliliter colloidal gold solution, add then in the 300-500ug immune protein IgG solution, at room temperature stirred 10-15 minute;
3. the 0.8-1.2% polyglycol solution that adds the 4-10 milliliter again;
4. restrain centrifugal actions 20-40 minute through 10000～100000, the careful suction removed supernatant;
5. sediment is mixed with the damping fluid of 0.2～0.5mg/ml polyglycol of 80-120 milliliter again;
6. repeat with 10000～100000 gram centrifugal actions 20-40 minute, the careful suction removed supernatant;
7. after sediment being mixed with the damping fluid of 0.2～0.5mg/ml polyglycol of 80-120 milliliter, add the Sodium azide of 30-70 milligram again, reached anticorrosion purpose after the mixing, putting temperature is 3-6 ℃ of preservation.
The invention provides a kind of method of making immune chromatograph testing strip, wherein: the molecular weight of described polyglycol is 20000; Described acetate is potassium acetate or sodium acetate; Protein solution is albumin solution or king crab haemocyanin.
The invention provides a kind of method of making immune chromatograph testing strip, wherein: reducing process is adopted in the preparation of described colloidal gold solution; Its step is as follows:
(a). at first glass container was soaked in the chloroformic solution of 3%-10% dichlorodimethylsilane 30 seconds to 1 minute, and used distilled water flushing after the drying at room temperature, again drying for standby;
(b). the 0.08%-0.12% aqueous solution of chloraurate of getting the 80-120 milliliter adds to be heated in the glass container that prior dried crosses and boils, the 0.8%-1.2% reductive agent aqueous solution that adds the 0.5-0.9 milliliter then while stirring, this mixed liquor gold just became purplish red solution by yellow solution in 2 minutes, continued to boil 10-20 minute, the cooling back adds distilled water makes the volume of solution return to the 80-120 milliliter, finishes the preparation of collaurum;
The invention provides a kind of method of making immune chromatograph testing strip, wherein: described reductive agent is sodium citrate, tannic acid, ascorbic acid, white phosphorus or sodium borohydride.
The present invention compared with prior art, the present invention is owing to utilize colloidal gold-labeled method and quantitative immune competition law to produce the test-strips of fast detecting, can quick and precisely detect the micro-hormone in the human body, the fast detection method that detects with test-strips of the present invention has specificity and sensitivity highly, also has good stability, repeatability by force, do not use specific apparatus, expense is cheap, easy easy and simple to handle, short time just can interpretation go out the result, is beneficial to people and detects the hormonal readiness of self at any time.
Description of drawings
Fig. 1 is the side schematic view of immune chromatograph testing strip of the present invention.
Embodiment
Referring to Fig. 1, immune chromatograph testing strip of the present invention comprises stilt 1, water-absorption fiber film 2, nitrocellulose membrane 3, sample film 4, waterproof membrane 5 and colloid gold label film 6, nitrocellulose membrane 3 sticks to the stage casing of bar shaped stilt 1, the water-absorption fiber film sticks to the top of stilt 1, the upper end overlap joint of the lower end of water-absorption fiber film 2 and nitrocellulose membrane 3, colloid gold label film 6, waterproof membrane 5 and sample film 4 stick to end from stilt 1 successively to stilt 1 nitrocellulose membrane 3, and an end of colloid gold label film 6 and waterproof membrane 5 overlap joints, one end of waterproof membrane 5 and sample film 4 overlap joints, one end of sample film 4 and nitrocellulose membrane 3 overlap joints, wherein the lap of splice of colloid gold label film 6 one ends and waterproof membrane 5 sticks to length on the stilt 1 greater than waterproof membrane 5, comprise three kinds of variable concentrations on the described nitrocellulose membrane 2 with detect corresponding antibody and a kind of anti-immune protein antibody of antigen (as progestational hormone antigen or estrogen antigen), described four kinds antibody longitudinally evenly distributes to absorbing membrane 2 ends from being positioned at sample film 4 ends, the spraying of every kind of antibody highly is the 1-3 millimeter, be spaced apart the 2-3 millimeter between four kinds of antibody, comprise with colloid gold label on the colloid gold label film 6, the antigen that is detected and with the immune protein IgG of colloid gold label, described sample film 4 and colloid gold label film 6 are glass fibre membrane, described detection antigen is micromolecule antigen, as: progestational hormone antigen or estrogen antigen.
The preparation method of immune chromatograph testing strip comprises the preparation method of nitrocellulose membrane 2 and the preparation method of colloid gold label film 6, wherein the preparation method of collaurum film 6 is added on the glass fibre membrane after mixing with the antigenic solution of colloid gold label with the immune protein solution of colloid gold label, makes the colloid gold label film after the drying.
The preparation of colloid gold label film 6
(1) preparation of colloidal gold solution
Its concrete steps are as follows:
(a). at first glass container was soaked in the chloroformic solution of 6% dichlorodimethylsilane 1 minute, and used distilled water flushing after the drying at room temperature, again drying for standby;
(b). get 90 milliliters 0.09% aqueous solution of chloraurate and add to be heated in the glass container that prior dried crosses and boil, 0.9% sodium citrate aqueous solution that adds 0.6 milliliter then while stirring, this mixed liquor gold just became purplish red solution by yellow solution in 2 minutes, continued to boil 15 minutes, the cooling back adds distilled water makes the volume of solution return to 90 milliliters, finishes the preparation of collaurum.
(2) preparation of the antigenic solution of colloid gold label
1. with micro-0.09mol/L liquor kalii acetici above-mentioned colloidal gold solution is adjusted to potential of hydrogen 8.3 earlier;
2. in 100 milliliters of colloidal gold solutions, add then in the albumin matter solution that will detect antigen of 350ug, at room temperature stirred 12 minutes;
3. 1% molecular weight that adds 7 milliliters again is 20000 polyglycol solutions;
4. restrain centrifugal actions 25 minutes through 10000～100000, the careful suction removed supernatant;
5. be that the damping fluid of 20000 polyglycol mixes with 90 milliliters 0.4mg/ml molecular weight again with sediment;
6. repeat with 10000～100000 gram centrifugal actions 25 minutes, the careful suction removed supernatant;
7. be after the damping fluid of 20000 polyglycol mixes with 90 milliliters 0.4mg/ml molecular weight again with sediment, add 45 milligrams Sodium azide, reached anticorrosion purpose after the mixing, putting temperature is 5 ℃ of preservations.
(3) preparation of the immune protein solution of colloid gold label
1. with micro-0.11mol/L acetate solution above-mentioned colloidal gold solution is adjusted to potential of hydrogen 7.9 earlier;
2. in 100 milliliters of colloidal gold solutions, add 350ug immune protein IgG solution then, at room temperature stirred 14 minutes;
3. 1% molecular weight that adds 5 milliliters again is 20000 polyglycol solutions;
4. restrain centrifugal actions 25 minutes through 10000～100000, the careful suction removed supernatant;
5. sediment is mixed with the damping fluid of 100 milliliters 0.4mg/ml polyglycol again;
6. repeat with 10000～100000 gram centrifugal actions 25 minutes, the careful suction removed supernatant;
7. after sediment being mixed with the damping fluid of 100 milliliters 0.4mg/ml polyglycol, add 50 milligrams Sodium azide again, reached anticorrosion purpose after the mixing, putting temperature is 4 ℃ of preservations.
(4) preparation of colloid gold label film
Be added on the glass fibre membrane after the immune protein solution of the above-mentioned colloid gold label of the antigenic solution of the above-mentioned colloid gold label of 50 microlitres and 50 microlitres mixed, room temperature or temperature be 4-5 ℃ dry down, make the colloid gold label film.
The preparation of nitrocellulose membrane
On described nitrocellulose membrane 2 different parts, spray three kinds of various dose respectively with the antigen reactive antibody of colloid gold label and a kind of anti-immune protein antibody, they are followed successively by spraying is detection line 1 with the antigen reactive antibody of colloid gold label for three kinds, 2,3 and a kind of anti-immune protein antibody be control line, described different parts be every kind of antibody near sample film 4 ends to the wire that is that sprays successively near water-absorption fiber film 2 ends, four kinds of dosage of antibody are respectively 0.4 microgram, 1.2 microgram, 3.0 microgram and 4.8 micrograms, highly about 1 mm wide of the spraying of every line 3 millimeters long, control line is positioned at apart from nitrocellulose filter and water-absorption fiber film junction 5 millimeters, each line is 2～3 mm distance at interval, spraying nitrocellulose filter 3 later is after drying, albumin solution with 50 grams per liters soaks sealing 2 hours, with 0.01 mole physiological solution washing 3 times, the sealing of dry back is preserved again.
The preparation method of immune chromatograph testing strip
Immune chromatograph testing strip is that nitrocellulose membrane 3 is sticked on the stage casing of bar shaped stilt 1, the water-absorption fiber film is sticked to the top of stilt 1, the upper end overlap joint of the lower end of water-absorption fiber film 2 and above-mentioned nitrocellulose membrane 3, with above-mentioned colloid gold label film 6, waterproof membrane 5 and sample film 4 stick to end from stilt 1 successively to stilt 1 nitrocellulose membrane 3, and an end of colloid gold label film 6 and waterproof membrane 5 overlap joints, one end of waterproof membrane 5 and sample film 4 overlap joints, one end of sample film 4 and nitrocellulose membrane 3 overlap joints, wherein the lap of splice of colloid gold label film 6 one ends and waterproof membrane 5 sticks to length on the stilt 1 greater than waterproof membrane 5, makes immune chromatograph testing strip.
The using method of immune chromatograph testing strip
Describe to detect estrogen below:
1. 0.1-0.5 milliliter saliva or urine are added on the sample film 4, wait 1 minute;
2. will remove at the waterproof membrane between sample film and the colloid gold label film 5;
3. 0.1 ml water is added on and contains on the colloid gold label film 6, wait 2 minutes;
4. the variation on the observation test bar nitrocellulose filter 3, detection line a red stripes occurs and represents that estrogen is that two red stripes of 400pg. are 200pg, article three, red stripes is that four red stripes of 50pg/ml. are that estrogen level is lower than 50pg/ml. and does not have red stripes to occur then being considered as estrogen level to be higher than 400pg/ml. under the test-strips normal condition, and control line should show red stripes.
The principle of work of immune chromatograph testing strip
To contain estrogenic saliva or urine adds on the sample film, saliva or urine are under the effect of absorbing membrane 2, be drawn on the nitrocellulose membrane 3, with 1 on the nitrocellulose membrane 3, antigen combination on 2 or 3 detection lines, after 1 minute, the user tears waterproof membrane, 0.1 ml water is added on contains on the colloid gold label film, water makes colloid gold label film 6 and sample film 4 stick together tightly, make on the colloid gold label film 6 with the estrogen antigen of colloid gold label and with the immune protein of colloid gold label under the effect of absorbing membrane 2, be drawn on the nitrocellulose membrane 3, with the estrogen antigen of colloid gold label and the antibody response colour developing that is neutralized by the estrogen in saliva or the urine fully on the nitrocellulose membrane, immune protein with colloid gold label develops the color after protein antibodies (being control line) combines with anti-the exempting from the nitrocellulose membrane, using the purpose of colloid gold label immune protein is to judge in order to point out the user whether to develop the color by the control line on the nitrocellulose membrane 3, whether test-strips lost efficacy, from the colour developing situation of observing nitrocellulose membrane 3 as can be seen: the estrogen that contains saliva or the urine is many more, the bar number of detection line colour developing is few more, so can carry out semiquantitative detection to the estrogen in saliva or the urine.
The foregoing description is just in order to make the public further understand the present invention, and the description of doing is not a limitation of the invention.
Claims (14)
1. immune chromatograph testing strip, it comprises stilt (1), water-absorption fiber film (2), nitrocellulose membrane (3), sample film (4), waterproof membrane (5) and colloid gold label film (6), it is characterized in that: nitrocellulose membrane (3) sticks to the stage casing of bar shaped stilt (1), the water-absorption fiber film sticks to the top of stilt (1), the upper end overlap joint of the lower end of water-absorption fiber film (2) and nitrocellulose membrane (3), colloid gold label film (6), waterproof membrane (5) and sample film (4) stick to end from stilt (1) successively to stilt (1) the nitrocellulose membrane (3), and an end of colloid gold label film (6) and waterproof membrane (5) overlap joint, one end of waterproof membrane (5) and sample film (4) overlap joint, one end of sample film (4) and nitrocellulose membrane (3) overlap joint, wherein the lap of splice of colloid gold label film (6) one ends and waterproof membrane (5) sticks to length on the stilt (1) greater than waterproof membrane (5), at least comprise a kind of pairing antibody of antigen with being detected of concentration and the antibody of a kind of anti-immune protein IgG on the described nitrocellulose membrane (2), comprise with colloid gold label on the colloid gold label film (6), the antigen that is detected and with the immune protein IgG of colloid gold label.
2. immune chromatograph testing strip according to claim 1 is characterized in that: the pairing antibody of the antigen with being detected that comprises three kinds of variable concentrations on the described nitrocellulose membrane (2).
3. immune chromatograph testing strip as claimed in claim 2, it is characterized in that: the antibody corresponding with detection antigen of described three kinds of concentration and the antibody of a kind of anti-immune protein IgG longitudinally evenly distribute to absorbing membrane (2) end from being positioned at sample film (4) end, the spraying of every kind of antibody highly is the 1-3 millimeter, is spaced apart the 2-3 millimeter between four kinds of antibody.
4. as immune chromatograph testing strip as described in the claim 3, it is characterized in that: described sample film (4) and colloid gold label film (6) are glass fibre membrane.
5. as the described immune chromatograph testing strip of one of claim 1 to 4, it is characterized in that: described detection antigen is micromolecule antigen.
6. immune chromatograph testing strip as claimed in claim 5 is characterized in that: described little antigen molecule antigen is progestational hormone antigen or estrogen antigen.
7. method of making immune chromatograph testing strip, it is characterized in that: it comprises nitrocellulose membrane (3) is sticked on the stage casing of bar shaped stilt (1), the water-absorption fiber film is sticked to the top of stilt (1), the upper end overlap joint of the lower end of water-absorption fiber film (2) and nitrocellulose membrane (3), with colloid gold label film (6), waterproof membrane (5) and sample film (4) stick to end from stilt (1) successively to stilt (1) the nitrocellulose membrane (3), and an end of colloid gold label film (6) and waterproof membrane (5) overlap joint, one end of waterproof membrane (5) and sample film (4) overlap joint, one end of sample film (4) and nitrocellulose membrane (3) overlap joint, wherein the lap of splice of colloid gold label film (6) one ends and waterproof membrane (5) sticks to length on the stilt (1) greater than waterproof membrane (5), at least apply a kind of pairing antibody of antigen with being detected and a kind of anti-immune protein antibody of concentration on the described nitrocellulose membrane (2), colloid gold label film (6) is to apply the antigenic solution that is detected indicate colloid gold label and with the immune protein solution of colloid gold label on glass fibre membrane.
8. the manufacture method of immune chromatograph testing strip as claimed in claim 7, it is characterized in that: on described nitrocellulose membrane (2) different parts, spray three kinds of various dose respectively with the antigen reactive antibody of colloid gold label and a kind of anti-immune protein antibody, described different parts is that every kind of antibody becomes to be wire from hold close water-absorption fiber film (2) end to spray successively near sample film (4), three kinds of various dose be followed successively by detection line 1 with the antigen reactive antibody of colloid gold label, 2,3, a kind of anti-immune protein antibody is control line, four kinds of dosage of antibody are respectively the 0.3-0.6 microgram, 1.0-1.7 microgram, 2.6-3.9 microgram and 4.4-5.6 microgram, highly about 1 mm wide of the spraying of every line 3 millimeters long, control line is positioned at apart from nitrocellulose filter and water-absorption fiber film junction 3-6 millimeter, each line is 2～3 mm distance at interval, spraying nitrocellulose filter (3) later is after drying, albumin solution with 40 grams per liters-60 grams per liter soaks sealing 1-3 hour, physiological solution with the 0.008-0.013 mole washs 2-4 time again, and the sealing of dry back is preserved.
9. as the manufacture method of claim 7 or 8 described immune chromatograph testing strips, it is characterized in that: being added on the glass fibre membrane after the immune protein solution with colloid gold label with the antigenic solution of colloid gold label and 20-60 microlitre of 20-60 microlitre is mixed, is that 3-6 ℃ of following drying made the colloid gold label film in room temperature or temperature.
10. the manufacture method of immune chromatograph testing strip as claimed in claim 9 is characterized in that: the manufacturing step of described antigenic solution with colloid gold label is as follows:
1. with micro-0.08-012mol/L acetate solution colloidal gold solution is adjusted to potential of hydrogen 8.2-8.6 earlier;
2. in 100 milliliters of colloidal gold solutions, add then in the protein solution that will detect antigen of 300-500ug, at room temperature stirred 10-15 minute;
3. the 0.8-1.3% polyglycol solution that adds the 4-10 milliliter again;
4. restrain centrifugal actions 20-40 minute through 10000～100000, the careful suction removed supernatant;
5. sediment is mixed with the damping fluid of 0.2～0.5mg/ml polyglycol of 80-120 milliliter again;
6. repeat with 10000～100000 gram centrifugal actions 20-40 minute, the careful suction removed supernatant;
7. after sediment being mixed with the damping fluid of 0.2～0.5mg/ml polyglycol of 80-120 milliliter, add the Sodium azide of 30-70 milligram again, reached anticorrosion purpose after the mixing, putting temperature is 3-6 ℃ of preservation.
11. the manufacture method of immune chromatograph testing strip as claimed in claim 9 is characterized in that: the manufacturing step of the immune protein solution of described usefulness colloid gold label is as follows:
1. with micro-0.08-012mol/L acetate solution colloidal gold solution is adjusted to potential of hydrogen 7.8-8.2 earlier;
2. in 100 milliliters of colloidal gold solutions, add 300-500ug immune protein IgG solution then, at room temperature stirred 10～15 minutes;
3. the 0.8-1.2% polyglycol solution that adds the 4-10 milliliter again;
4. restrain centrifugal actions 20-40 minute through 10000～100000, the careful suction removed supernatant;
5. sediment is mixed with the damping fluid of 0.2～0.5mg/ml polyglycol of 80-120 milliliter again;
6. repeat with 10000～100000 gram centrifugal actions 20-40 minute, the careful suction removed supernatant;
7. after sediment being mixed with the damping fluid of 0.2～0.5mg/ml polyglycol of 80-120 milliliter, add the Sodium azide of 30-70 milligram again, reached anticorrosion purpose after the mixing, putting temperature is 3-6 ℃ of preservation.
12. the manufacture method as claim 10 or 11 described immune chromatograph testing strips is characterized in that: the molecular weight of described polyglycol is 20000; Described acetate is potassium acetate or sodium acetate; Described protein solution is albumin solution or king crab haemocyanin.
13. the manufacture method of immune chromatograph testing strip as claimed in claim 12 is characterized in that: reducing process is adopted in the preparation of described colloidal gold solution; Its step is as follows:
(a). at first glass container was soaked in the chloroformic solution of 3%-10% dichlorodimethylsilane 30 seconds to 1 minute, and used distilled water flushing after the drying at room temperature, again drying for standby;
(b). the 0.08%-0.12% aqueous solution of chloraurate of getting the 80-120 milliliter adds to be heated in the glass container that prior dried crosses and boils, the 0.8%-1.2% reductive agent aqueous solution that adds the 0.5-0.9 milliliter then while stirring, this mixed liquor gold just became purplish red solution by yellow solution in 2 minutes, continued to boil 10-20 minute, the cooling back adds distilled water makes the volume of solution return to the 80-120 milliliter, finishes the preparation of collaurum.
14. the manufacture method of immune chromatograph testing strip as claimed in claim 13 is characterized in that: described reductive agent is sodium citrate, tannic acid, ascorbic acid, white phosphorus or sodium borohydride.
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| CNB2004100745222A CN1311239C (en) | 2004-09-07 | 2004-09-07 | Immune chromatograph testing strip and production thereof |
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