CN103336113A - Preparation method of human immunodeficiency virus (HIV) antibody detection test paper - Google Patents
Preparation method of human immunodeficiency virus (HIV) antibody detection test paper Download PDFInfo
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- CN103336113A CN103336113A CN2013102518951A CN201310251895A CN103336113A CN 103336113 A CN103336113 A CN 103336113A CN 2013102518951 A CN2013102518951 A CN 2013102518951A CN 201310251895 A CN201310251895 A CN 201310251895A CN 103336113 A CN103336113 A CN 103336113A
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Abstract
The invention discloses a preparation method of human immunodeficiency virus (HIV) antibody detection test paper. The preparation method comprises the following steps of: spraying gold on a gold cushion by a mixed solution of a P74 recombinant antigen mark and a P44 recombinant antigen mark through membrane marking and gold spraying equipment to obtain a first gold cushion; spraying gold on the gold cushion by a rabbit IgG monoclonal antibody mark to obtain a second gold cushion; marking on a polyvinyl chloride bottom plate stuck with a nitrocellulose membrane by a detection line covering solution and a control line covering solution to obtain a polyvinyl chloride bottom plate with a marked membrane; assembling a sample cushion, the first gold cushion, the second gold cushion, the polyvinyl chloride bottom plate with the marked membrane and water absorption paper; and cutting the assembly into the detection test paper. In the manner, the detection test paper prepared by the preparation method disclosed by the invention can be used for judging whether an oral cavity sample contains an HIV antibody or not based on an immune lateral flow chromatography technology through manual operation and naked eye reading; the diagnosis is rapid, the result is accurate, and damages to a body of a subject are not caused.
Description
Technical field
The present invention relates to the biologic applications technical field, particularly relate to a kind of preparation method of HIV (human immunodeficiency virus) antibody test test paper.
Background technology
Human immunodeficiency virus (Human Immunodeficiency Virus, HIV) be a kind of slow virus of infected person para-immunity system cells, belong to a kind of of retroviruse, be the mortality infectious disease of not having effective therapy so far, extensively be present in the infected's blood, seminal fluid, vaginal fluid, saliva, urine, milk, cerebrospinal fluid and have in the brain tissue liquid of nervous symptoms.Human immunodeficiency virus can be destroyed the immunocompetence of human body, causes immune system to lose resistibility, thereby causes various diseases and cancer to be survived in human body, develops into to cause acquired immune deficiency syndrome (AIDS) at last.
For acquired immune deficiency syndrome (AIDS), make a definite diagnosis in early days and be conducive to patient and obtain medical treatment as early as possible, also be beneficial to the control transmission of disease, therefore diagnosis is particularly important as early as possible.Detection to acquired immune deficiency syndrome (AIDS) at present is divided into blood testing and does not have the wound detection, is that feminine gender or the positive judge whether to fall ill by testing result.Blood testing will puncture person under inspection's skin blood sample collection, and this wound type checking can bring misery to patient, also can increase the infected danger of medical worker.Not having that wound detects is to analyze the mouth cavity liquid of collecting to obtain diagnostic result, but the method wastes time and energy, and needs to wait for for a long time the result.
Summary of the invention
The technical matters that the present invention mainly solves provides a kind of preparation method of HIV (human immunodeficiency virus) antibody test test paper, this method system detection paper accurate, harmless.
For solving the problems of the technologies described above, the technical scheme that the present invention adopts is: the preparation method that a kind of HIV (human immunodeficiency virus) antibody test test paper is provided, comprise that step is: (1) is water-soluble and constant volume with trishydroxymethylaminomethane, sucrose, trehalose and bovine serum albumin(BSA), regulate the pH value to 8-9, obtain golden indicated weight suspension, the pH value of regulating the 0.01M disodium phosphate soln with the 0.01M sodium dihydrogen phosphate obtains bag and is cushioned liquid to 7-8;
(2) in the collaurum that stirs, add 0.2M sal tartari, regulate pH, add thing to be marked and stirring, add the 0.1g/mL bovine serum albumin solution again, the centrifugal precipitation that obtains, described precipitation is resuspended to 1/10 of centrifugal front volume with described golden indicated weight suspension, and wherein said thing to be marked is P74 recombinant antigen, P44 recombinant antigen or rabbit igg monoclonal antibody, obtains P74 recombinant antigen mark, P44 recombinant antigen mark or rabbit igg monoclonal antibody mark;
(3) be cushioned liquid dilution P57 recombinant antigen with bag and the P36 recombination antigen mixture obtains the detection line coating buffer, be cushioned liquid with bag and dilute the how anti-dilution of goat anti-rabbit igg and obtain the control line coating buffer;
(4) with the mixed liquor of described P74 recombinant antigen mark and described P44 recombinant antigen mark gold pad metal spraying is obtained the first gold medal pad by selecting film metal spraying equipment, with described rabbit igg monoclonal antibody mark gold pad metal spraying is obtained the second gold medal pad, rule at the Polyvinylchloride base plate that is stained with nitrocellulose filter respectively with described detection line coating buffer and described control line coating buffer, obtain the Polyvinylchloride base plate of a film;
(5) Polyvinylchloride base plate and the thieving paper with sample pad, the first gold medal pad, the second gold medal pad, some film fits together, and cuts into HIV (human immunodeficiency virus) antibody test test paper.
In a preferred embodiment of the present invention, the ratio of trishydroxymethylaminomethane, described sucrose, described trehalose, described bovine serum albumin(BSA) and constant volume cumulative volume is 0.24g:20g:5g:1g:100mL described in the step (1).
In a preferred embodiment of the present invention, the pH value of golden indicated weight suspension is adjusted to 8.5 with 1M hydrochloric acid described in the step (1), and the pH value that described bag is cushioned liquid is adjusted to 7.4.
In a preferred embodiment of the present invention, collaurum was filtration described in the step (2), described stirring is to place on the magnetic stirring apparatus, adds 0.2M sal tartari in the collaurum that stirs, and regulates pH and stirs 5min, add thing to be marked and stir 60min, adding the 0.1g/mL bovine serum albumin solution again, stir 30min, is under the condition of 11000r/min centrifugal 60 minutes at 4 ℃, rotating speed, precipitation is resuspended to 1/10 of centrifugal front volume with described golden indicated weight suspension, obtains label.
In a preferred embodiment of the present invention, the ratio of 0.2M sal tartari and described P74 recombinant antigen described in the step (2) is 9.5uL/mL:16 μ g/mL, the ratio of described 0.2M sal tartari and described P44 recombinant antigen is 9.5uL/mL:8 μ g/mL, the ratio of described 0.2M sal tartari and described rabbit igg monoclonal antibody is 8uL/mL:10 μ g/mL, and the addition of described bovine serum albumin(BSA) accounts for 1% of cumulative volume.
In a preferred embodiment of the present invention, the concentration of P57 recombinant antigen described in the detection line coating buffer is 1.6mg/mL described in the step (3), the concentration of described P36 recombinant antigen is 1.6mg/mL, and how anti-concentration is 2mg/mL to goat anti-rabbit igg described in the described control line coating buffer.
In a preferred embodiment of the present invention, the pad of gold described in the step (4) adopts glass fibre membrane, before metal spraying, soak behind the 10min dry with gold pad treating fluid, wherein said gold pad treating fluid is that the dodecyl polyglycol ether is added to the water constant volume and obtains, and the ratio of described dodecyl polyglycol ether and constant volume cumulative volume is 1g:100mL.
In a preferred embodiment of the present invention, sample pad described in the step (5) adopts glass fibre membrane, soak behind the 10min dryly before use with the sample pad treating fluid, wherein said sample pad treating fluid is to be that potassium dihydrogen phosphate, trishydroxymethylaminomethane, sodium hydrogen phosphate, thimerosal, lauryl sodium sulfate, ethylenediamine tetraacetic acid and the bovine serum albumin(BSA) constant volume soluble in water of 0.2g:6.06g:1.15g:0.1g:1.25g:1g:10g is to the 1L volume with ratio.
The invention has the beneficial effects as follows: the preparation method of HIV (human immunodeficiency virus) antibody test test paper of the present invention, the detection test paper that obtains is based on immune lateral flow chromatographic technique, can read to judge whether contain HIV antibody in the sample of oral cavity by manual operations, naked eyes, namely can read the result after 20 ~ 30 minutes, at the latest can be above 45 minutes, diagnostic result is accurate, can not damage person under inspection's health.
Embodiment
Below preferred embodiment of the present invention is described in detail, thereby so that advantages and features of the invention can be easier to be it will be appreciated by those skilled in the art that protection scope of the present invention is made more explicit defining.
(1) container and material
Tankage: beaker, graduated cylinder, volumetric flask, glass bar, magnetic stirring apparatus, electronic balance, liquid-transfering gun, some film metal spraying equipment, guillotine, cutting cutter, capper, case pressing machine, pH meter, constant temperature vacuum drying chamber.
Reagent auxiliary material: gold chloride, trisodium citrate, KCl, K
2CO
3, the Tris(trishydroxymethylaminomethane), HCl, BSA(bovine serum albumin(BSA)), the EDTA(ethylenediamine tetraacetic acid), lauryl sodium sulfate, T-20, Brij-35(dodecyl polyglycol ether), sucrose, trehalose, Sodium azide.
Material: glass fibre membrane, polyester film, NC film, base plate, thieving paper, drying agent, aluminium foil bag, plastic clip, tygon and polyester synthetic material.
Environment: cleaning, room temperature, humidity is below 60%, water quality: process water.
(2) preparation process
1, the preparation of collaurum: adopt trisodium citrate reduction method
The preparation of 100 mL, 1% chlorauric acid solution: take by weighing 1 g gold chloride and be dissolved in the process water, volumetric flask is settled to 100 mL, and 4 ℃ of preservations are standby.
The preparation of 100 mL, 1% citric acid three sodium solution: take by weighing 1 g trisodium citrate and be dissolved in the process water, volumetric flask is settled to 100 mL, filters with 0.22 μ m filter, and is now with the current.
520nm Preparation of Colloidal Gold method: in clean beaker, add process water, on magnetic stirring apparatus, be heated to and boil, add 1% citric acid three sodium solution then, fully mixing 2min adds 1% chlorauric acid solution, keeps heating 8min, volume is recovered in the cooling back, and 4 ℃ of preservations are standby.
Each component adds volume:
2, the pre-service of sample pad, gold pad
1L sample pad treating fluid preparation: take by weighing 0.2g potassium dihydrogen phosphate, 6.06g Tris, 1.15g sodium hydrogen phosphate, 0.1g thimerosal, 1.25g lauryl sodium sulfate, 1g EDTA and 10g bovine serum albumin(BSA) and be dissolved in the process water, volumetric flask is settled to 1L, and room temperature preservation is standby.
The preparation of 100 mL gold pad treating fluid: take by weighing 1g Brij-35 and be dissolved in the process water, volumetric flask is settled to 100 mL, and 4 ℃ of preservations are standby.
The gold pad that with material is glass fibre membrane RB65 soaks in gold pad treating fluid, takes out after soaking 10min, is set to bone dry in 37 ℃ the vacuum drying chamber in temperature, cuts out the trimming edge and prevents edge effect, and the aluminium foil bag room temperature preservation is standby.
The sample pad that with the material model is SX42 is soaked in the sample pad treating fluid, soaks 15min, takes out, and is set to bone dry in 37 ℃ the vacuum drying chamber in temperature, cuts out the trimming edge and prevents edge effect, and the aluminium foil bag room temperature preservation is standby.
3, golden indicated weight suspension, bag are cushioned the liquid preparation
The preparation of 100mL 1M hydrochloric acid: measure 8.59 mL concentrated hydrochloric acids and be dissolved in the process water, volumetric flask is settled to 100 mL, and room temperature preservation is standby.
The preparation of 100mL gold indicated weight suspension: take by weighing 0.24g Tris, 20g sucrose, 5g trehalose and 1g BSA and be dissolved in process water, constant volume in the 100mL volumetric flask, transferring pH with 1M hydrochloric acid is 8.5, filters with 0.22 μ m filter, 4 ℃ of preservations are standby.
Bag is cushioned the liquid preparation: take by weighing 0.12 g sodium hydrogen phosphate and be dissolved in process water, volumetric flask is settled to 100 mL, is the 0.01M disodium phosphate soln; Take by weighing 0.14 g sodium dihydrogen phosphate and be dissolved in process water, volumetric flask is settled to 100 mL, is the 0.01M sodium dihydrogen phosphate; Regulate the pH to 7.4 of 0.01 M disodium phosphate soln with 0.01 M sodium dihydrogen phosphate, the bag that is 0.01M, pH=7.4 is cushioned liquid, filters with 0.22 μ m filter, and 4 ℃ of preservations are standby.
4, immuno-gold labeling
The preparation of 100 mL, 0.2 M solution of potassium carbonate: take by weighing 2.76 g sal tartari and be dissolved in process water, volumetric flask is settled to 100 mL, filters with 0.22 μ m filter, and room temperature preservation is standby.
The preparation of 100 mL 10%BSA solution: take by weighing 10 g BSA and be dissolved in the process water, 100 mL volumetric flasks are settled to 100 mL, and 0.22 μ m filter filters standby.
P74 recombinant antigen mark: get beaker, add the collaurum that has filtered, put on the magnetic stirring apparatus and stir; the 0.2M sal tartari that adds 9.5uL/mL is regulated pH and is stirred 5min, slowly adds 16 μ g/mL P74 recombinant antigens; stir 60 min; slowly add 10% BSA protective agent, to final concentration 1%, stir 30 min; under the condition that 4 ℃ of following rotating speeds are 11000 r/min centrifugal 60 minutes; abandoning supernatant precipitates standby with resuspended 1/10, the 4 ℃ of preservation to original volume of golden indicated weight suspension.
P44 recombinant antigen mark: get beaker, add the collaurum that has filtered, put on the magnetic stirring apparatus and stir; the 0.2M sal tartari that adds 9.5uL/mL is regulated pH and is stirred 5min, slowly adds 8 μ g/mL P44 recombinant antigens; stir 60 min; slowly add 10% BSA protective agent, to final concentration 1%, stir 30 min; under the condition that 4 ℃ of following rotating speeds are 11000 r/min centrifugal 60 minutes; abandoning supernatant precipitates standby with resuspended 1/10, the 4 ℃ of preservation to original volume of golden indicated weight suspension.
Rabbit igg monoclonal antibody mark: get beaker, add the collaurum that has filtered, put on the magnetic stirring apparatus and stir; the 0.2M sal tartari that adds 8uL/mL is regulated pH and is stirred 5min, slowly adds 10 μ g/mL rabbit igg monoclonal antibody; stir 60 min; slowly add 10% BSA protective agent, to final concentration 1%, stir 30 min; under the condition that 4 ℃ of following rotating speeds are 11000 r/min centrifugal 60 minutes; abandoning supernatant precipitates standby with resuspended 1/10, the 4 ℃ of preservation to original volume of golden indicated weight suspension.
5, some film metal spraying operation
Metal spraying: open a film metal spraying equipment, the metal spraying module is subdued, the line module carries on, and prevents that scribe head from touching the plate face, opens vacuum pump to 0.5 atmospheric pressure.
Enter the operation picture by [operation], select [program] to be transformed into the metal spraying mode of operation.Choose program number after entering the routine data picture, edit ' length ', ' concentration ', ' speed ', ' parameters such as initial point X, Y, Z ', revised parameter after, preserve data by [preservations], will forever preserve data by [permanent preservation].
Enter manual picture by [manually], in No. 2 pumps (metal spraying pattern) sample pipe insertion process water test tube, pipe end is prepared the vessel of water receiving.Select [cleaning] to use process water to clean 3 times.After waiting to take the vessel of water receiving away, select [discharge opeing] in pipeline, not have moisture.Insert No. 2 pump sample pipes in the test tube this moment, is full of in pipeline in the mixed liquor or rabbit igg monoclonal antibody mark of P74 recombinant antigen mark and P44 recombinant antigen mark by [liquid feeding].The gold pad of handling well is placed on correct position on the plate face, enters the operation picture by [operation], press [beginning] and carry out metal spraying.
Metal spraying finishes, and with in No. 2 pump sample pipe insertion process water test tubes, pipe end is prepared the vessel of water receiving.Select [cleaning] to use process water to clean 3 times.Opening gas to air pressure by [the gas operation] of [master menu] is zero.
Gold pad after metal spraying is handled in vacuum drying chamber at 37 ℃ of following vacuum drying 60min to bone dry.Knot gold pad after dry the processing places aluminium foil bag, adds the drying agent room temperature and preserves standbyly, and what P74 recombinant antigen mark and P44 recombinant antigen mark mixing metal spraying obtained is the first gold medal pad, and what obtain with rabbit igg monoclonal antibody mark metal spraying is the second gold medal pad.
Preparation detection line, control line coating buffer: being cushioned liquid dilution P57 recombinant antigen to final concentration with bag is 1.6 mg/mL, dilution P36 recombinant antigen to final concentration is 2.4 mg/mL, and the two mixing is obtained the detection line coating buffer is T line working fluid, and 4 ℃ of preservations are standby.
How anti-to be diluted to final concentration be 2 mg/mL with goat anti-rabbit igg to be cushioned liquid with bag, and obtaining the control line coating buffer is C line working fluid, and 4 ℃ of preservations are standby.
The point film: the module of will ruling is subdued, and the metal spraying module is carried on, and forms certain drop, prevents metal spraying head Contact plate face.
Enter the operation picture by [operation], select [program] to be transformed into router's operation mode.Choose program number after entering the routine data picture, edit ' length ', ' concentration ', ' speed ', ' parameters such as initial point X, Y, Z ', wherein ' speed ' can not be too fast, be 100 mm/s, after having revised parameter, preserve data by [preservation], will forever preserve data by [the permanent preservation].
Enter manual picture by [manually], in 1, No. 3 pump (line pattern) sample pipe insertion process water test tube, pipe end is prepared the vessel of water receiving.Select [cleaning] to use process water to clean 3 times.After waiting to take the vessel of water receiving away, select [discharge opeing] in pipeline, not have moisture.Insert No. 1 pump (line pattern) sample pipe in the detection line coating buffer this moment, and No. 3 pumps (line pattern) sample pipe is inserted in the control line coating buffer, is full of coating buffer by [liquid feeding] in pipeline.The ready PVC base plate that is pasted with nitrocellulose filter is placed on correct position on the plate face, enters the operation picture by [operation], press [beginning] and rule.
The line end of operation, with in 1, No. 3 pump sample pipe insertion process water test tube, pipe end is prepared the vessel of water receiving.Select [cleaning] to use process water to clean 3 times, close close point film metal spraying instrument.
The Polyvinylchloride base plate that will be stained with nitrocellulose filter after line is finished places drying box, and dry 30min is to bone dry under 37 ℃.After drying is finished described Polyvinylchloride base plate is placed aluminium foil bag, add the drying agent room temperature and preserve standby.
6, assembling cutting, assembling, packing
PVC base plate and the absorbing membrane of sample pad, the first gold medal pad, the second gold medal pad, some film are fitted together, use automatic cutting machine to cut into the 4mm test strips.Every test strips is assembled in the plastic clip, uses case pressing machine to compress.Add in the aluminium foil bag of packing into behind the drying agent and preserve, be finished product.
Described HIV (human immunodeficiency virus) antibody test test paper has been fixed Recombinant HIV-1 and HIV-2 type antigen at the detection zone of cellulose acetate membrane, fixed the goat anti-rabbit igg segment in the check plot how anti-, wraps by rabbit igg-collaurum and Recombinant HIV-1 and HIV-2 type antigen-collaurum in advance on glass fibre membrane.During test, with buccal swab wiping experimenter's upper and lower gum line, then detecting device is put in the damping fluid bottle that fills an amount of sample buffer, arrived the nitrocellulose filter watch window until sample buffer.Take out kit, horizontal positioned, Recombinant HIV-1 and HIV-2 type antigen-colloid gold label thing on the gold pad can dissolve, with the HIV-1 in the sample, 2 type antibody combine, form " antibody-antigen-Jin " compound, compound is rearward mobile along test strips, at first arrives bag by HIV detection of antigens district.If contain the antibody of HIV in the sample, " antibody-antigen-Jin " compound will be with the envelope antigen combination, and under detection line is detained, forms a pink line, and this represents positive findings.The depth of line color is with the disproportional relation of antibody quantity in the sample.If there are not colored lines to represent not contain in the sample HIV antibody in the detection zone, free " antibody-antigen-Jin " compound continues to move to the test paper rear, arrives the check plot of test paper.The check plot bag is fixed on the control line of test strips by the goat anti-rabbit igg segment." rabbit igg-Jin " compound will combine and will be enriched on the control line with the goat anti-rabbit igg segment, form a pink line.The appearance proof test strips measuring ability of control line is normal.No matter whether contain HIV-1/2 antibody in the sample, in efficiency test, light red control line to purple all should appear in the check plot of test strips.Sample continues mobile, enters the uptake zone by the check plot.The effect of uptake zone is absorption remaining " antibody-antigen-Jin " and sample composite thing, makes it mobile in test strips, and eliminates the color of background.Sample mixed liquor timing after test strips is inserted dilution, the rear can read the result in 20 ~ 30 minutes, but at the latest must not be above 45 minutes.
The above only is embodiments of the invention; be not so limit claim of the present invention; every equivalent structure or equivalent flow process conversion that utilizes description of the present invention to do; or directly or indirectly be used in other relevant technical fields, all in like manner be included in the scope of patent protection of the present invention.
Claims (8)
1. the preparation method of a HIV (human immunodeficiency virus) antibody test test paper, it is characterized in that, comprise that step is: (1) is water-soluble and constant volume with trishydroxymethylaminomethane, sucrose, trehalose and bovine serum albumin(BSA), regulate the pH value to 8-9, obtain golden indicated weight suspension, the pH value of regulating the 0.01M disodium phosphate soln with the 0.01M sodium dihydrogen phosphate obtains bag and is cushioned liquid to 7-8;
(2) in the collaurum that stirs, add 0.2M sal tartari, regulate pH, add thing to be marked and stirring, add the 0.1g/mL bovine serum albumin solution again, the centrifugal precipitation that obtains, described precipitation is resuspended to 1/10 of centrifugal front volume with described golden indicated weight suspension, and wherein said thing to be marked is P74 recombinant antigen, P44 recombinant antigen or rabbit igg monoclonal antibody, obtains P74 recombinant antigen mark, P44 recombinant antigen mark or rabbit igg monoclonal antibody mark;
(3) be cushioned liquid dilution P57 recombinant antigen with bag and the P36 recombination antigen mixture obtains the detection line coating buffer, be cushioned liquid with bag and dilute the how anti-dilution of goat anti-rabbit igg and obtain the control line coating buffer;
(4) with the mixed liquor of described P74 recombinant antigen mark and described P44 recombinant antigen mark gold pad metal spraying is obtained the first gold medal pad by selecting film metal spraying equipment, with described rabbit igg monoclonal antibody mark gold pad metal spraying is obtained the second gold medal pad, rule at the Polyvinylchloride base plate that is stained with nitrocellulose filter respectively with described detection line coating buffer and described control line coating buffer, obtain the Polyvinylchloride base plate of a film;
(5) Polyvinylchloride base plate and the thieving paper with sample pad, the first gold medal pad, the second gold medal pad, some film fits together, and cuts into HIV (human immunodeficiency virus) antibody test test paper.
2. preparation method according to claim 1 is characterized in that, the ratio of trishydroxymethylaminomethane, described sucrose, described trehalose, described bovine serum albumin(BSA) and constant volume cumulative volume is 0.24g:20g:5g:1g:100mL described in the step (1).
3. preparation method according to claim 1 is characterized in that, the pH value of golden indicated weight suspension is adjusted to 8.5 with 1M hydrochloric acid described in the step (1), and the pH value that described bag is cushioned liquid is adjusted to 7.4.
4. preparation method according to claim 1, it is characterized in that, collaurum was filtration described in the step (2), described stirring is to place on the magnetic stirring apparatus, in the collaurum that stirs, add 0.2M sal tartari, regulate pH and stir 5min, add thing to be marked and stir 60min, add the 0.1g/mL bovine serum albumin solution again, stir 30min, be under the condition of 11000r/min centrifugal 60 minutes at 4 ℃, rotating speed, precipitation obtains label with 1/10 of the resuspended extremely centrifugal front volume of described golden indicated weight suspension.
5. preparation method according to claim 1, it is characterized in that, the ratio of 0.2M sal tartari and described P74 recombinant antigen described in the step (2) is 9.5uL/mL:16 μ g/mL, the ratio of described 0.2M sal tartari and described P44 recombinant antigen is 9.5uL/mL:8 μ g/mL, the ratio of described 0.2M sal tartari and described rabbit igg monoclonal antibody is 8uL/mL:10 μ g/mL, and the addition of described bovine serum albumin(BSA) accounts for 1% of cumulative volume.
6. preparation method according to claim 1, it is characterized in that, the concentration of P57 recombinant antigen described in the detection line coating buffer is 1.6mg/mL described in the step (3), the concentration of described P36 recombinant antigen is 2.4mg/mL, and how anti-concentration is 2mg/mL to goat anti-rabbit igg described in the described control line coating buffer.
7. preparation method according to claim 1, it is characterized in that, the pad of gold described in the step (4) adopts glass fibre membrane, before metal spraying, soak behind the 10min dry with gold pad treating fluid, wherein said gold pad treating fluid is that the dodecyl polyglycol ether is added to the water constant volume and obtains, and the ratio of described dodecyl polyglycol ether and constant volume cumulative volume is 1g:100mL.
8. preparation method according to claim 1, it is characterized in that, sample pad described in the step (5) adopts glass fibre membrane, soak behind the 10min dryly before use with the sample pad treating fluid, wherein said sample pad treating fluid is to be that potassium dihydrogen phosphate, trishydroxymethylaminomethane, sodium hydrogen phosphate, thimerosal, lauryl sodium sulfate, ethylenediamine tetraacetic acid and the bovine serum albumin(BSA) constant volume soluble in water of 0.2g:6.06g:1.15g:0.1g:1.25g:1g:10g is to the 1L volume with ratio.
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Cited By (8)
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| CN103558381A (en) * | 2013-11-06 | 2014-02-05 | 昆明云大生物技术有限公司 | Immunochromatographic test paper for detecting human immunodeficiency virus antibodies and preparation method thereof |
| CN104678098A (en) * | 2015-03-13 | 2015-06-03 | 苏州万木春生物技术有限公司 | Preparation method of joint-inspection test paper for cardiac troponin I and heart-type fatty acid binding protein |
| CN105738622A (en) * | 2015-10-23 | 2016-07-06 | 北京玛斯玛克生物科技有限公司 | Human urine HIV1/2 antibody detection test paper through colloidal gold chromatographic method and preparation method thereof |
| CN106290849A (en) * | 2015-05-13 | 2017-01-04 | 上海凯创生物技术有限公司 | A kind of meningitis bacterium gold-immunochromatographyreagent reagent for assay box |
| CN106526173A (en) * | 2016-10-31 | 2017-03-22 | 广州科方生物技术股份有限公司 | A coating liquid used for immunochromatography test paper strips and a preparing method thereof |
| CN107014994A (en) * | 2016-12-23 | 2017-08-04 | 苏州万木春生物技术有限公司 | A kind of preparation method of antibody of HCV Test paper |
| CN110058012A (en) * | 2019-04-08 | 2019-07-26 | 石家庄洹众生物科技有限公司 | The fluorescence immunoassay test paper group and its preparation and application of synchronous detection diacetylmorphine, crystal methamphetamine, ketamine and cocaine |
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| CN110058012A (en) * | 2019-04-08 | 2019-07-26 | 石家庄洹众生物科技有限公司 | The fluorescence immunoassay test paper group and its preparation and application of synchronous detection diacetylmorphine, crystal methamphetamine, ketamine and cocaine |
| CN110058012B (en) * | 2019-04-08 | 2022-03-01 | 石家庄洹众生物科技有限公司 | Fluorescent immune test paper group for synchronously detecting diacetylmorphine, methamphetamine, ketamine and cocaine and preparation and use methods thereof |
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