CN102192986B - Cardiac troponin I detection reagent and preparation method thereof and application thereof - Google Patents
Cardiac troponin I detection reagent and preparation method thereof and application thereof Download PDFInfo
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- CN102192986B CN102192986B CN201010127883.4A CN201010127883A CN102192986B CN 102192986 B CN102192986 B CN 102192986B CN 201010127883 A CN201010127883 A CN 201010127883A CN 102192986 B CN102192986 B CN 102192986B
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- 101710128251 Troponin I, cardiac muscle Proteins 0.000 title abstract 4
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Abstract
The invention discloses a cardiac troponin I detection reagent and a preparation method thereof and an application thereof. The cardiac troponin I detection reagent is characterized in that: the detection reagent contains a nano-gold particle wrapped by organosilicon, wherein a particle size of the nano-gold particle is 10 to 150 nm and the thickness of the nano-silica coating is 5 to 100 nm; functional groups of amidine, carboxyl, hydroxy, hydrosulfuryl, cyclopropyl, aldehyde group are introduced into the surface of the organosilicon coating; and a biomolecule such as protein or antibody is marked by the nano gold with the functionalized organosilicon surface by chemical reaction so as to form a stable covalent bond connection and to maintain the biological activity of the marked biomolecule. The modified nano-gold particle, which can substitute for a traditional colloidal gold particle, can be used in the immunology field to prepare immunization reagent; and the modified nanometer colloid can be used to prepare the cardiac troponin I detection reagent.
Description
Technical field
The present invention relates to detection reagent of a kind of cardiac muscle troponin I (TROPONIN I) and its production and use, belong to biological medicine and detect reagent field.
Background technology
Angiocardiopathy is the first major disease that threatens human life's health.According to World Health Organization's recent statistics, the whole world has 1,670 ten thousand people to die from all kinds of cardiovascular and cerebrovascular diseases every year, and this numeral accounts for 29.2% of all disease deaths, wherein has the acute myocardial infarction AMI of dying from over half.In China, over nearly 10 years, the incidence of disease of China's acute myocardial infarction AMI obviously rises, and has approached international average level.Incidence of CHD accounts for 6 ‰ left and right of total population, and actual patients with coronary heart disease number reaches 7,800,000 people.And owing to lacking the general knowledge of preventing and treating of coronary heart disease, a lot of patients reach an advanced stage when medical, taking acute myocardial infarction AMI (AMI) as onset symptoms.
At present both at home and abroad to AMI diagnostic criteria be clinical symptoms as pectoralgia, electrocardiographic abnormality and serum cardiac biochemical marker are abnormal.But at the AMI initial stage, approximately 25% patient does not have typical clinical symptoms, patient's cardiogram of about half does not have typical ANOMALOUS VARIATIONS, and the blood sample mesophytization mark while therefore detecting the damage of AMI initial stage heart of patient has important references meaning extremely in AMI clinical diagnosis.
In China's situation of all-level hospitals, the main biochemical markers of heart detecting in laboratory comprises: TnT roponin-I(TNI), Troponin-T(TNT), myoglobins Myoglobin and cretinephosphokinase CK-MB.Wherein CK-MB is present in cardiac muscle in a large number, generally within 4-8 hour, raises in AMI morbidity, within 12-20 hour, peaks, and has been acknowledged as " goldstandard " that detect AMI since early seventies.But CK-MB specificity is poor, marathon race, Skeletal muscle injury, renal failure person CK-MB content all can increase extremely.The time that CK-MB exists in blood is shorter (1-4 days) also.Along with the development of detection technique, it is the most special that cardiac muscle troponin I (CTnI) has been proved to be myocardial damage, one of the most responsive blood serum designated object.In normal human blood, the content of CTnI is generally lower than 0.4ng/ml, and after AMI occurs, CTnI can raise fast in 6-8 hour, reaches peak value, and all maintained higher concentration in 18-24 hour in 10-15 days.Clinical fast detecting to CTnI has important society and economic benefit.
Existing CTnI detection method mainly contains euzymelinked immunosorbent assay (ELISA) (ELISA, chemoluminescence method and immunofluorescence technique.The lowest detection sensitivity of ELISA can reach 0.2ng/mL, and determination period is long, reaches 5-6 hour.Chemoluminescence method is luminesceence analysis and immune response to be combined and the trace antigen set up or the labelled immune analytical technology of antibody, its sensitivity is up to 0.03ng/mL, and detection time is also shorter, about 1-2 hour, but need to adopt special main equipment, complicated operation.Immunofluorescence technique adopts method sandwich and that enzyme connection fluorescence combines, and detection time is very fast, and maximum sensitivity can reach 0.1ng/mL, also needs specialized equipment.These detection methods all need expensive instrument and professional's operation, be only applicable to use in centralab, time in testing result arrival doctor hand is longer, can not meet clinical fast detecting needs, and being not suitable for middle and small hospital does not especially have the township hospital of centralab yet.
The kit of domestic existing fast detecting CTnI mainly adopts traditional colloidal gold immunity chromatography preparation.Its ultimate principle is the monoclonal antibody that adopts colloid gold label individual plant or the anti-CTnI of many strains, adopts the sandwich CTnI of detection of dual anti-immunity, Chinese patent 200710022815.x, 200810036418.2 existing reports.On market all based on CTnI quick detection kit detection sensitivity generally within the scope of 2-10ng/ml.
Collaurum refers to the suspending liquid of golden particulate, and its diameter, at 1～100nm, has high electron density, dielectric property and catalytic action, can be combined with multiple biomacromolecule, and not affect its biologically active.Its color takes on a red color to purple according to diameter.The building-up process of tradition collaurum, its mark to monoclonal antibody, the preparation of solid-phase immunity chromatograph test strip all has report.In the 1980s, first collaurum is applied to mark and the vitro detection field of protein molecular.
Collaurum is electronegative under weak base environment, can form firmly and be combined with the positive charge group of protein molecule, this combination is electrostatical binding, although do not affect the biological nature of protein, also more stable, but this combination is subject to solution ion strength, pH impact is very large, especially in high inonic strength solution, collaurum condenses, become unstable by the biomolecule of nano gold mark also temporal evolution, affected the mark of collaurum to a lot of biomolecule and the application in biological sample.
Summary of the invention
The object of the invention is provides detection reagent of a kind of cardiac muscle troponin I and its production and use for the deficiencies in the prior art, is characterized in synthesizing new nanoparticle, nano-particle surface functionalization, and the labeling method of nanoparticle antagonist.It also comprises reagent strip, contains the two anti-of the anti-cardiac muscle troponin I monoclonal antibody of anti-cardiac muscle troponin I monoclonal antibody and modified Nano gold mark and anti-cardiac muscle troponin I monoclonal antibody on reagent strip.
Object of the present invention is realized by lower technical measures: wherein said raw material umber, except specified otherwise, is parts by weight.
Detection reagent of cardiac muscle troponin I and preparation method thereof:
The detection reagent of cardiac muscle troponin I
The core particle diameter of organosilicon parcel nm of gold particulate is 10～150nm, be preferably 50～100nm, most preferably be 55～75nm, organosilicon surface coating is 5-100nm, amino, carboxyl, hydroxyl, sulfhydryl, trimethylene base and aldehyde radical functional group are introduced in described organic silicon coating surface, the organosilicon surface of functionalization, is formed stable covalent bond and connects as protein, antibody carry out mark by chemical reaction and biomolecule, and keeps being labeled the biologically active of molecule.
The preparation method that cardiac muscle troponin I detects reagent comprises the following steps:
1. the preparation of traditional collaurum
Be 100 parts of heating of 0.01～4% chlorauric acid solution by concentration, under stirring, boils 10～120 minutes 100～400rpm, adding fast concentration is 2～10 parts of 0.1～5 part of 2N ammonia spirit and the concentration sodium citrates that is 1%, high-speed stirred to solution presents orange red, cooling rear high speed centrifugation purifying, obtain the collaurum particulate of particulate footpath at 10～200nm, and the collaurum packing making is preserved;
2. the preparation of the nm of gold particulate of organosilicon parcel
By 100 parts of the above-mentioned colloidal gold solutions making, add 0.1～2 part of APTES, stir 15～120 minutes at 100～400rpm, add 200～500 parts of solvents, to alkalescence, add 0.1～2 part of tetraethoxy-silicane alkylating mixture by ammoniacal liquor regulator solution pH value, at room temperature continue to stir 18-24 hour, product, at 2000～10000rpm high speed centrifugation purifying, is obtained to the nm of gold particulate of organosilicon parcel;
3. organosilicon parcel nm of gold is surface-functionalized
100 parts of the above-mentioned collaurums making are added to 200～500 parts of solvents, stir 15～120 minutes at 100～400rpm, by extremely alkalescence of ammoniacal liquor adjusting pH value, add 0.1～2 part of tetraethoxysilane of 1: 1 and the mixed solution containing the silylating reagent of organic functional group, at room temperature continue to stir 18～24 hours, product, at 2000～10000rpm high speed centrifugation purifying, obtains the nanoparticle of the surface-functionalized modification of organic-capping nm of gold;
4. the mark of the nm of gold of organosilicon parcel to cTnI monoclonal antibody
Adopt surperficial carboxylated organosilicon parcel nm of gold activate its surperficial carboxyl by ethyl-3-(3-dimethyl aminopropyl)-carbodiimides (EDC) and react with the amino of cardiac muscle troponin I monoclonal antibody Fc, form the amide structure of stable chemical performance, this stable chemical bond is not subject to the ionic strength of solution, Acidity of Aikalinity affects, thereby increases chemistry and the storage stability of tagged cardiac muscle troponin I monoclonal antibody.Cardiac muscle troponin I monoclonal antibody good mark is sprayed on glass fibre pad, forms gold mark pad.
5. the preparation of kit
Using the monoclonal antibody of a strain CTnI as detecting the antibody phosphate buffer solution that antibody compound concentration is 1.5mg/ml, add 1～20 part of 2% trehalose, spray printing, to the nitrocellulose filter of activation, forms detection line; Anti-to nitrocellulose filter, form quality control line as sheep anti-mouse igg spray printing by two of the monoclonal antibody of CTnI; By dry nitrocellulose filter 2-10 hour, preparation detects the nitrocellulose filter of CTnI.By gold mark pad, sample pad, nitrocellulose filter and adsorptive pads stick on plastic plate, plastic plate is cut into 5mm width detection CTnI test strips, then pack CTnI test strip into plastic casing, make the kit that detects CTnI.
6. the method for quick of cardiac muscle troponin I
Whole blood/blood serum sample is added to the sample well on kit, be penetrated in sample pad, in sample, the monoclonal detection antibody of the CTnI of CTnI and organosilicon parcel nano gold mark meets in conjunction with forming compound, compound continues to be penetrated into detection line and the mixed binding that is fixed on CTnI monoclonal on detection line and catches antibody and form sandwich double antibody along nitrocellulose filter, the redness that shows nm of gold on detection line, testing result is positive; The CTnI monoclonal of unnecessary nano gold mark detects antibody in the time that quality control line is arrived in dialysis, with the sandwich combination of two anti-formation being fixed on quality control line, make nature controlling line also show redness, show that the dialysis of sample on nitrocellulose filter is complete, this testing result is effective.If CTnI content is lower than detection sensitivity in sample, the CTnI monoclonal of nano gold mark detects antibody can not form effective sandwich mixed binding with fixing seizure antibody on detection line, detection line can not show redness, testing result is negative, but the CTnI monoclonal of nano gold mark detect antibody still can with the sandwich combination of the anti-formation of monoclonal two being fixed on nature controlling line, on nature controlling line, show redness, show that testing result is effective.This method is technology well known to those skilled in the art.
Organic functions group is any in amino, carboxyl, sulfhydryl, hydroxyl, trimethylene base or aldehyde radical functional group.
Solvent is any in ethanol, isopropyl alcohol, methyl alcohol, normal butyl alcohol.
The nm of gold particulate of the surface-functionalized modification of organosilicon parcel nm of gold carries out mark by chemical reaction and biomolecule, forms stable covalence key and connects.
Detect the content of reagent for the cardiac muscle troponin I of clinical fast detecting infraction blood.
Tool of the present invention has the following advantages:
1, the present invention adopts organosilicon to wrap up traditional nm of gold particulate, forms the silicon coating of nano thickness on its surface, and chemical modification is carried out in silicon coating surface, introduces carboxyl, amino, hydroxyl, sulfhydryl, trimethylene base and aldehyde radical functional group.The collaurum that organic-silicon-modified nm of gold is more traditional has better stability, is not easy to occur the cohesion of gold grain.Silicon face introduce chemical functional group and biomolecule as albumen, antibody, antigen forms stablizes chemical covalent bonds, prepares more stable nano gold mark thing.The nm of gold of this modification can replace traditional collaurum for field of immunology, prepares immunoreagent.
2, utilize this nm of gold to bioprotein, antibody carries out mark, between biomolecule and nm of gold particulate, forms and stablizes covalent bond, has improved the chemistry and the storage stability that are labeled molecule.
3, surface silicon coating can improve the surface reflection light intensity of nano material, thereby improves the naked eyes resolution of this material, and the sensitivity that has improved immunoreagent, can reach 0.4ng/mL.
4, the labeling of monoclonal antibody of the nm of gold of utilizing this organosilicon parcel to cardiac muscle troponin I also prepared the detection kit that detects cardiac muscle troponin I by immunochromatographic method, greatly improved the detection speed of cardiac muscle troponin I.
Embodiment
Below by embodiment, the present invention is specifically described; be necessary to be pointed out that at this present embodiment is only used to further illustrate the present invention; can not be interpreted as limiting the scope of the invention, the person skilled in the art in this field can make some nonessential improvement and adjustment according to the content of the invention described above.
Embodiment 1
1: preparation and the surface amination of organosilicon parcel nanogold particle
The preparation of nanogold particle solution: adopt sodium citrate reducing process to prepare collaurum particulate solution, the particle size of collaurum is controlled at 10-150nm.100 parts of the gold chloride deionized water solutions that is 1% by concentration, high-speed stirred is also boiled 10 minutes.Adding fast concentration is 2 parts of 0.1 part of the hydrogen amine aqueous solution of 2N and sodium citrates that concentration is 1%, high-speed stirred, solution presents orange red, the collaurum particulate that the particle diameter that cooling rear high speed centrifugation purifying obtains particulate is 55-75nm, the collaurum obtaining is preserved a period of time and then organosilicon parcel aminated modification is carried out in collaurum surface, 100 parts of the colloidal gold solutions of obtaining, add 300 parts of ethanol, stir 15 minutes at 100～400rpm, by extremely alkalescence of ammoniacal liquor regulator solution pH, the tetraethoxysilane and the APTES mixed solution that add 1 part of 1:1 to mix, at room temperature continue to stir 18-24 hour, product is at 2000～10000rpm high speed centrifugation purifying, obtain the novel nano gold particulate of organosilicon parcel surface amination.
The anti-cardiac muscle troponin I monoclonal antibody of 2 surface amination organosilicon parcel nano gold mark
The organosilicon of preparing in the method for 100 parts of foregoing descriptions wraps up in the aminated nano-Au solution in surface, add the anti-cardiac muscle troponin I monoclonal antibody of 10 parts of crosslinking chemicals and 100 micrograms to continue reaction 2 hours, then adding concentration is 0.1 part of the phosphate buffer of 10% bovine serum albumin(BSA), continues reaction 1 hour.After high speed centrifugation purifying, obtain the anti-cardiac muscle troponin I monoclonal antibody of surface amination organosilicon parcel nano gold mark.
The preparation of 3 cardiac muscle troponin I kits
The monoclonal antibody of the anti-cardiac muscle troponin I of the modified Nano gold mark of above-mentioned preparation is sprayed to and on the glass fibre scraps of paper, prepares the immune modified Nano gold scraps of paper.The monoclonal of cardiac muscle troponin I is caught to antibody and two anti-IgG(1.5mg/mL with specking instrument) on the detection line and nature controlling line of specking on nitrocellulose filter, be dried 2 hours at temperature 37oC, acquire immune cellulose nitrate film strip, thieving paper is pasted on immune cellulose nitrate film strip, and cut into the wide test strips of 5mm, by the test strips of cutting, golden millimeter paper and sample pad, filter pad is assembled in sample box, makes the kit of cardiac muscle troponin I.
Embodiment 2
Preparation and the surface of 1 organosilicon parcel nanogold particle are carboxylated
Prepare by the described method of embodiment 1 the collaurum particulate that particle diameter is 55-75nm, get above-mentioned 100 parts of the colloidal gold solutions that make, add 250 parts of ethanol, stir 65 minutes at 100～400rpm, by extremely alkalescence of ammoniacal liquor regulator solution pH, add 1 part of tetraethoxysilane, react 2 hours, add again 1 part of maleamic acid propyl-triethoxysilicane, at room temperature continue to stir 18-24 hour, product, at 2000～10000rpm high speed centrifugation purifying, obtains the carboxylated novel nano gold particulate in organosilicon parcel surface.
The anti-cardiac muscle troponin I monoclonal antibody of 2 surperficial carboxylated organosilicon parcel nano gold mark
The organosilicon of preparing in the method for 100 parts of foregoing descriptions wraps up in the carboxylated nano-Au solution in surface, the concentration that adds 5 parts of 1:1 to mix is that 20mmol1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides (EDC) and concentration are 20mmol N-hydroxy-succinamide (NHS) mixed solution, at room temperature react 45 minutes, add the anti-cardiac muscle troponin I monoclonal antibody of 50 microgram to continue reaction 2 hours, then adding concentration is 0.1 part of the phosphate buffer of 10% bovine serum albumin(BSA), continues reaction 1 hour.After high speed centrifugation purifying, obtain the anti-cardiac muscle troponin I monoclonal antibody of surperficial carboxylated organosilicon parcel nano gold mark.
The preparation of 3 cardiac muscle troponin I kits
The monoclonal antibody of the anti-cardiac muscle troponin I of the modified Nano gold mark of above-mentioned preparation is sprayed to and on the glass fibre scraps of paper, prepares the immune modified Nano gold scraps of paper.The monoclonal of cardiac muscle troponin I is caught to antibody and two anti-IgG(1.5mg/mL with specking instrument) on the detection line and nature controlling line of specking on nitrocellulose filter, be dried 2 hours at temperature 37oC, acquire immune cellulose nitrate film strip, thieving paper is pasted on immune cellulose nitrate film strip, and cut into the wide test strips of 5mm, by the test strips of cutting, golden millimeter paper and sample pad, filter pad is assembled in sample box, makes the kit of cardiac muscle troponin I.
Embodiment 3
1: preparation and the Surface Hydrogen sulfation of organosilicon parcel nanogold particle
Prepare by the described method of embodiment 1 the collaurum particulate that particle diameter is 55-75nm, get 100 parts of the above-mentioned colloidal gold solutions making, add 200 parts of isopropyl alcohols, stir 120 minutes at 100～400rpm, by extremely alkalescence of ammoniacal liquor regulator solution pH, add part 0.1 part of tetraethoxysilane and 3-sulfenyl propyl-triethoxysilicane mixed solution that 1:1 mixes, at room temperature continue to stir 18-24 hour.Product, at 2000～10000rpm high speed centrifugation purifying, obtains the novel nano gold particulate of organosilicon parcel Surface Hydrogen sulfation.
The anti-cardiac muscle troponin I monoclonal antibody of 2 Surface Hydrogen sulfation organosilicon parcel nano gold mark
By adding 5 microgram activators in the phosphate buffer of 100 microgram cardiac muscle troponin I monoclonal antibodies, under room temperature, react 20 minutes, with gel chromatographic columns purification reaction thing, obtain the cardiac muscle troponin I monoclonal antibody solution of activation.The nano-Au solution of the organosilicon parcel of describing in 1 part of embodiment 3 will be added in the monoclonal antibody of activation, under room temperature, react 1 hour, adding concentration is 0.1 part of the phosphate buffer of 5% bovine serum albumin(BSA), continue reaction 1 hour, after high speed centrifugation purifying, obtain the anti-cardiac muscle troponin I monoclonal antibody of Surface Hydrogen sulfation organosilicon parcel nano gold mark, between the antibody of mark and silicon parcel nm of gold, form stable chemical covalent bond and be connected, increased chemical stability and the storage stability of labelled antibody.
The preparation of 3 cardiac muscle troponin I kits
The monoclonal antibody of the anti-cardiac muscle troponin I of the modified Nano gold mark of above-mentioned preparation is sprayed to and on the glass fibre scraps of paper, prepares the immune modified Nano gold scraps of paper.The monoclonal of cardiac muscle troponin I is caught to antibody and two anti-IgG(1.5mg/mL with specking instrument) on the detection line and nature controlling line of specking on nitrocellulose filter, be dried 2 hours at temperature 37oC, acquire immune cellulose nitrate film strip, thieving paper is pasted on immune cellulose nitrate film strip, and cut into the wide test strips of 5mm, by the test strips of cutting, golden millimeter paper and sample pad, filter pad is assembled in sample box, makes the kit of cardiac muscle troponin I.
Embodiment 4
1: preparation and the surperficial trimethylene base of organosilicon parcel nanogold particle
Prepare by the described method of embodiment 1 the collaurum particulate that particle diameter is 55-75nm, get above-mentioned 100 parts of the colloidal gold solutions that make, add 250 parts of ethanol, stir 75 minutes at 100～400rpm, by extremely alkalescence of ammoniacal liquor regulator solution pH, add 1 part of tetraethoxysilane, react 2 hours, add again 1 part of (3-glycidoxy propyl group) dimethylethoxysilane, at room temperature continue to stir 18-24 hour, product is at 2000～10000rpm high speed centrifugation purifying, and acquisition organosilicon wraps up the novel nano gold particulate of surperficial trimethylene base.
The anti-cardiac muscle troponin I monoclonal antibody of 2 surperficial trimethylene base organosilicon parcel nano gold mark
The organosilicon of preparing in the method for 100 parts of foregoing descriptions wraps up in the nano-Au solution of surperficial propyl, add the anti-cardiac muscle troponin I monoclonal antibody of 50 microgram to continue reaction 2 hours at 38oC, then adding concentration is 0.1 part of the phosphate buffer of 10% bovine serum albumin(BSA), continues reaction 1 hour.After high speed centrifugation purifying, obtain the anti-cardiac muscle troponin I monoclonal antibody of surperficial trimethylene base organosilicon parcel nano gold mark.
The preparation of 3 cardiac muscle troponin I kits
The monoclonal antibody of the anti-cardiac muscle troponin I of the modified Nano gold mark of above-mentioned preparation is sprayed to and on the glass fibre scraps of paper, prepares the immune modified Nano gold scraps of paper.The monoclonal of cardiac muscle troponin I is caught to antibody and two anti-IgG(1.5mg/mL with specking instrument) on the detection line and nature controlling line of specking on nitrocellulose filter, be dried 2 hours at temperature 37oC, acquire immune cellulose nitrate film strip, thieving paper is pasted on immune cellulose nitrate film strip, and cut into the wide test strips of 5mm, by the test strips of cutting, golden millimeter paper and sample pad, filter pad is assembled in sample box, makes the kit of cardiac muscle troponin I.
Claims (6)
1. the detection reagent of a cardiac muscle troponin I, it is characterized in that the nm of gold particulate that this detection reagent contains organosilicon parcel, the core particle diameter of this particulate is 10～150nm, organosilicon surface coating is 5～100nm, described organosilicon surface coating is introduced amino, carboxyl, hydroxyl, sulfhydryl, trimethylene base and aldehyde radical functional group, the organosilicon surface coating of functionalization carries out mark by chemical reaction and biomolecule, form stable covalent bond and connect, and keep being labeled the biologically active of molecule;
And prepare by following processing step and technological parameter:
(1) preparation of the nm of gold particulate of organosilicon parcel and surface-functionalized
Be 0.01～4% chlorauric acid solution 100 weight portion heating by concentration, under stirring, boils 10～120 minutes 100～400rpm, adding fast concentration is sodium citrate 2～10 weight portions that 2N ammonia spirit 0.1～5 weight portion and concentration are 1%, high-speed stirred to solution presents orange red, cooling rear high speed centrifugation purifying, obtain the collaurum particulate of diameter of particle at 10～150nm, and the collaurum packing making is preserved; Again by the above-mentioned colloidal gold solution making 100 weight portions, add 200～500 parts by weight solvent, stir 15～120 minutes at 100～400rpm, by extremely alkalescence of ammoniacal liquor regulator solution pH value, add 0.1～2 weight portion tetraethoxysilane of 1: 1 and the mixed solution containing the silylating reagent of functional group, at room temperature continue to stir 18～24 hours, product, at 2000～10000rpm high speed centrifugation purifying, is obtained to the nanoparticle of the surface-functionalized modification of organosilicon parcel nm of gold;
(2) preparation of the detection reagent of cardiac muscle troponin I
Adopt dual anti-immune sandwich method that the cardiac muscle troponin I monoclonal of the above-mentioned nanoparticle mark making is detected to antibody specking on glass fibre pad, another cardiac muscle troponin I monoclonal catches two anti-speckings of antibody and monoclonal detection antibody in nitrocellulose filter, form the Rapid detection test strip of anti-cardiac muscle troponin I, then by this test strip generate a reagent box.
2. the detection reagent of cardiac muscle troponin I as claimed in claim 1, is characterized in that the core particle diameter of the nm of gold particulate of organosilicon parcel is 50～100nm.
3. the detection reagent of cardiac muscle troponin I as claimed in claim 1, is characterized in that the core particle diameter of the nm of gold particulate of organosilicon parcel is 55-75nm.
4. the detection reagent of cardiac muscle troponin I as claimed in claim 1, is characterized in that biomolecule is protein or antibody.
5. the detection reagent of cardiac muscle troponin I as claimed in claim 1, is characterized in that solvent is any in ethanol, isopropyl alcohol, methyl alcohol, normal butyl alcohol.
6. the detection reagent of cardiac muscle troponin I as claimed in claim 1, is characterized in that containing the silylating reagent of functional group be any in APTES, maleamic acid propyl-triethoxysilicane, 3-sulfenyl propyl-triethoxysilicane or (3-glycidoxy propyl group) dimethylethoxysilane.
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| EP3258270B1 (en) * | 2015-02-10 | 2019-05-29 | Shenzhen New Industries Biomedical Engineering Co. Ltd. | Cardiac troponin i ultra-sensitive detection reagent kit, and ultra-sensitive detection method therefor |
| CN109187981A (en) * | 2018-08-01 | 2019-01-11 | 万东山 | A kind of quantum dot immune chromatograph test strip of quick detection beta-amyloid protein and application |
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