CN104297483A - Quantum dot immunochromatography test strip for synchronous and quantitative joint inspection of HBsAg, HBeAg and HBcAb and method for synchronous and quantitative joint inspection of HBsAg, HBeAg and HBcAb - Google Patents
Quantum dot immunochromatography test strip for synchronous and quantitative joint inspection of HBsAg, HBeAg and HBcAb and method for synchronous and quantitative joint inspection of HBsAg, HBeAg and HBcAb Download PDFInfo
- Publication number
- CN104297483A CN104297483A CN201410544442.2A CN201410544442A CN104297483A CN 104297483 A CN104297483 A CN 104297483A CN 201410544442 A CN201410544442 A CN 201410544442A CN 104297483 A CN104297483 A CN 104297483A
- Authority
- CN
- China
- Prior art keywords
- hbeag
- hbsag
- quantum dot
- cds
- zns
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis
- G01N33/5761—Hepatitis B
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Communicable Diseases (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention particularly relates to a quantum dot immunochromatography test strip for synchronous and quantitative joint inspection of HBsAg, HBeAg and HBcAb and a method for the synchronous and quantitative joint inspection of the HBsAg, the HBeAg and the HBcAb. A label pad (3) of the test strip is coated with a mixture of a rat anti-HBsAg monoclonal antibody, a rat anti-HBeAg monoclonal antibody and HBcAg of corresponding labels of quantum dots with different wavelengths; a T belt (4) of an analyzing membrane (7) is coated with a mixture of a rabbit anti-HBsAg antibody, a rabbit anti-HBeAg antibody and the HBcAg; and a C belt (5) of the analyzing membrane (7) is coated with a goat anti-rat antibody. An HBsAg, HBeAg and HBcAb standard curve is stored and mounted on the test strip by an electronic label. The test strip adopts a detector with a signal detection function to read the standard curve stored by the electronic label and combines corresponding fluorescence intensity of a sample to be detected, which is measured by the detector, so as to synchronously and quantitatively detect the concentrations of the HBsAg, the HBeAg and the HBcAb in a blood sample.
Description
Technical field
The invention belongs to in-vitro diagnosis field, be specifically related to a kind of quantum dot immune chromatography examination bar and method thereof of synchronous quantitatively joint inspection hepatitis B three (HBsAg, HBeAg, HBcAb).
Background technology
Hepatitis B is the serious infectious diseases of serious harm human health, and China is hepatitis B infected big country, has people more than 600,000,000 to infect hepatitis B, about 1.3 hundred million people's Hepatitis B carriers, and hepatitis B control has extremely important meaning.At present, Serum Markers of Hepatitis B Virus still can not accomplish that many index quantitatively detects simultaneously and rapidly.Tradition enzyme linked immunosorbent assay (ELISA) can only detect a kind of index at every turn, perform repeatedly Parallel testing flow process and finally could obtain each Indexs measure result, analysis time long (often do an ELISA detection and at least need 40-60 minute), reagent consumption is many, expensive, complicated operation, labor capacity is large.
Quantum dot (quantum dot, QD) fluorescence radiation efficiency is high, exciting line wide ranges, energy " elementary excitation; polynary transmitting ", spectral line of emission narrow range and symmetrical, photobleaching speed is slow, and fluorescence lifetime is long, particle diameter is close with biomolecule, energy multifunction after finishing, the characteristic wavelength fluorescence spectrum that the quantum dot potpourri of different-grain diameter and kind produces is not overlapping, is highly suitable for sample multicomponent analysis.The present invention discloses a kind of quantum dot immune chromatography examination bar and method thereof of synchronous quantitatively joint inspection hepatitis B three, to solve Serum Markers of Hepatitis B Virus multi objective quantitative test problems simultaneously and rapidly.
Summary of the invention
First object of the present invention is the quantum dot immune chromatography examination bar of openly a kind of synchronous quantitatively joint inspection hepatitis B three.Second object of the present invention is the preparation method of openly described examination bar, standard curve making method and synchronously quantitatively detect HBsAg(hepatitis B surface antigen with described examination bar), HBeAg(hepatitis B virus e antigen), HBcAb(hepatitis B core antibody) method.
Above-mentioned purpose of the present invention is achieved by the following technical solution:
The quantum dot immune chromatography examination bar of described synchronous quantitatively joint inspection hepatitis B three, comprises sample pad 1, red blood cell filter membrane 2, label pad 3, analyzing film 7, adsorptive pads 6 that overlap joint is in turn fixed on end liner 8.
Described label pad 3 is glass fibre membrane.Described analyzing film 7 is nitrocellulose filter, nylon membrane or cellulose nitrate/cellulose acetate hybrid films, it has detection zone (i.e. T band) 4 and quality control band (i.e. C band) 5.Described end liner 8 is polyester or plastic plate.
Described label pad 3 is coated with the potpourri of the mouse-anti HBsAg monoclonal antibody of different wave length quantum dot correspondence markings, mouse-anti HBeAg monoclonal antibody and HBcAg.The T band 4 of described analyzing film 7 is coated with the potpourri of rabbit AntiHBsAg antibody, the anti-HBeAg antibody of rabbit, HBcAg.The C band 5 of described analyzing film 7 is coated with two anti-Quality Control thing sheep anti-mouse antibodies.
The quantum dot of the label pad 3 of examination bar of the present invention comprises ZnS, CdS, HgS, ZnSe, CdSe, HgSe, CdTe, ZnTe, ZnO, PbSe, HgTe, CaAs, InP, InAs, InCaAs, CdS/ZnS, CdS/Ag
2s, CdS/PbS, CdS/Cd (0H)
2, CdS/HgS, CdS/HgS/CdS, ZnS/CdS, ZnS/CdS/ZnS, ZnS/HgS/ZnS/CdS, CdSe/CdS, CdSe/ZnS, CdSe/ZnSe, CdSe/CuSe, CdSe/HgTe, CdSe/HgSe, CdSe/HgSe/CdSe, CdTe/HgS, CdTe/HgTe, InAs/InP, InAs/CdSe, InAs/ZnSe, MgS, MgSe, MgTe, CaS, CaSe, CaTe, SrS, SrSe, SeTe, BaS, BaSe, BaTe, CdS:Mn, ZnS:Mn, CdS:Cu, ZnS:Cu, CdS:Tb, the combination of any one or any several nano particle in ZnS:Tb, and be core by any one quantum dot above-mentioned, silicon dioxide is the core-shell type nano-complex particle of shell.
The preparation method of the quantum dot immune chromatography examination bar of synchronous quantitatively joint inspection hepatitis B three of the present invention comprises the steps:
A. quantum point coupling antibody/antigen:
a)getting different wave length quantum dot uses phosphate buffer (PBS) to adjust pH=6-9 respectively, adds EDC(l-(3-dimethyl aminopropyl)-3-ethyl carbon diamine hydrochloride) and NHS(N-hydroxy succinimide) room temperature activation 10-60min.
b)correspondence adds mouse-anti HBsAg monoclonal antibody, mouse-anti HBeAg monoclonal antibody and HBcAg vortex oscillating reactions 0.5-3h respectively.
c)correspondence adds BSA capping 0.5-2h respectively.
d)centrifugal purification product.
e)get the resuspended dispersion of precipitation PBS damping fluid, 4 DEG C of preservations.
B. bar assembly preparation is tried:
a)sample pad 1: select cellulose membrane to be cut into the film block of certain specification, soaked by the PBS that this film block is put into containing 0.1%-10% BSA and 0.01%-10% Tween 20, takes out, drying for standby.
b)red blood cell filter membrane 2: select red blood cell filter membrane to be cut into certain specification film block, drying for standby.
c)label pad 3: select glass fibre membrane to be cut into certain specification film block, add the mixture solution of the mouse-anti HBsAg monoclonal antibody by different wave length quantum dot correspondence markings, mouse-anti HBeAg monoclonal antibody and HBcAg on this film block, desciccator diaphragm block is for subsequent use.
d)analyzing film 7: select cellulose membrane to be cut into the film block of certain specification, from film block base, make T by the lower potpourri from the upper 0.5-10mg/ml of specking respectively rabbit AntiHBsAg antibody separated by a distance, the anti-HBeAg antibody of 0.5-10mg/ml rabbit, 0.5-10mg/ml HBcAg be with 4, specking 0.5-10mg/ml bis-anti-Quality Control thing sheep anti-mouse antibody makes C band 5, and desciccator diaphragm block is for subsequent use.
e)adsorptive pads 6: select the cellulose membrane with water sorption to be cut into the film block of certain specification, drying for standby.
C. bar assembly assembling is tried:
The above-mentioned examination bar assembly prepared overlaps in turn by sample pad 1, red blood cell filter membrane 2, label pad 3, analyzing film 7, adsorptive pads 6 and is pasted on end liner 8, is cut into the examination bar of certain specification, loads in plastic casing, loads sealed storage in aluminium foil bag together with drying agent.
The preparation method that examination bar of the present invention is used for the synchronous quantitatively typical curve of joint inspection hepatitis B three (HBsAg, HBeAg, HBcAb) comprises the steps:
(a)prepare each standard items series concentration of HBsAg, HBeAg, HBcAb respectively, and be added drop-wise on the quantum dot immune chromatography examination bar of described synchronous quantitatively joint inspection hepatitis B three.
(b)fluorescence intensity (the OD of T band is read respectively with detector
t) and C band fluorescence intensity (OD
c), calculate and obtain OD
t/ OD
cratio or OD
t/ (OD
t+ OD
c) ratio.
(c)x-axis is made, OD with standard items series concentration
t/ OD
cratio makes Y-axis, or makes X-axis with standard items series concentration, OD
t/ (OD
t+ OD
c) ratio makes Y-axis, obtains fluorescence intensity typical curve corresponding to concentration.
The method that examination bar of the present invention is used for synchronous quantitatively joint inspection hepatitis B three (HBsAg, HBeAg, HBcAb) comprises the steps:
(a)by electronic tag storage typical curve data.Described electronic tag comprises the RFID(RFID tag with information storage function), Quick Response Code, bar code or IC card chip.
(b)storage has the electronic tag of typical curve data to be arranged on quantum dot immune chromatography examination bar, or is arranged on the examination barrel of loading examination bar.
(c)with there are the typical curve data of detector reading electronic labels storage of signal testing function and fluorescence intensity corresponding to the testing sample recorded in conjunction with detector and HBsAg, HBeAg, HBcAb concentration of obtaining in sample.
The present invention has following beneficial effect:
(1) application of sample can realize sample HBsAg, HBeAg, HBcAb and quantitatively detect simultaneously and rapidly.
(2) red blood cell filter membrane 2 is provided with between the sample pad 1 of described examination bar and label pad 3, this red blood cell filter membrane 2 can stop the red blood cell in blood sample pass through and its serum can only be allowed to filter, blood sample does not need, with separation of serum such as centrifugation apparatus, just can directly detect with whole blood.
Accompanying drawing explanation
Fig. 1: the structure side view of the quantum dot immune chromatography examination bar of synchronous quantitatively joint inspection hepatitis B three (HBsAg, HBeAg, HBcAb) of the present invention
Fig. 2: the typical curve that examination bar of the present invention is used for synchronous quantitatively joint inspection HBsAg, HBeAg, HBcAb and obtains
Fig. 3: the examination bar T that examination bar of the present invention is used for synchronous quantitatively joint inspection HBsAg, HBeAg, HBcAb and obtains is with fluorescence spectrum figure
Sequence number and symbol are expressed as follows:
1. sample pad, 2. red blood cell filter membrane, 3. label pad, 4. detection zone, 5. quality control band, 6. adsorptive pads, 7. analyzing film, 8. end liner, T: detection zone (Test), C: quality control band (Control), Ag: antigen, McAb: monoclonal antibody.
Embodiment
embodiment 1: synchronous quantitatively joint inspectionhBsAg, HBeAg, HBcAb
quantum dot immune chromatography examination bar structure
Composition graphs 1 is explained.In Fig. 1, the quantum dot immune chromatography examination bar of described synchronous quantitatively joint inspection hepatitis B three, it comprises sample pad 1, red blood cell filter membrane 2, label pad 3, analyzing film 7, adsorptive pads 6 that overlap joint is in turn fixed on end liner 8.
The label pad 3 of described examination bar is glass fibre membrane.Described analyzing film 7 is nitrocellulose filter, nylon membrane or cellulose nitrate/cellulose acetate hybrid films, it has detection zone (i.e. T band) 4 and quality control band (i.e. C band) 5.Described end liner 8 is polyester or plastic plate.
Described label pad 3 is coated with CdSe/ZnS QD
526mouse-anti HBsAg monoclonal antibody (the Mouse anti-HBsAg of mark
mcAb-CdSe/ZnS QD
526), CdSe/ZnS QD
615mouse-anti HBeAg monoclonal antibody (the Mouse anti-HBeAg of mark
mcAb-CdSe/ZnS QD
615) and CdSe/ZnS QD
566hBcAg(and the CdSe/ZnS QD of mark
566-HBcAg) potpourri.The T band 4 of described analyzing film 7 is coated with the potpourri of rabbit AntiHBsAg antibody (Rabbit anti-HBsAg), the anti-HBeAg antibody of rabbit (Rabbit anti-HBeAg) and HBcAg.The C band 5 of described analyzing film 7 is coated with two anti-Quality Control thing sheep anti-mouse antibodies.
embodiment 2: synchronous quantitatively joint inspectionhBsAg, HBeAg, HBcAb
quantum dot immune chromatography examination bar preparation method
The preparation method of described examination bar comprises the steps:
A. quantum point coupling antibody/antigen:
a)get the water-soluble quantum dot CdSe/ZnS QD that emission wavelength is 526nm, 615nm and 566nm
526, CdSe/ZnS QD
615, CdSe/ZnS QD
566adjust pH=6-9 with PBS damping fluid respectively, add EDC(l-(3-dimethyl aminopropyl)-3-ethyl carbon diamine hydrochloride respectively) and NHS(N-hydroxy succinimide) room temperature activation 10-60min;
b)add mouse-anti HBsAg monoclonal antibody (Mouse anti-HBsAg respectively
mcAb), mouse-anti HBeAg monoclonal antibody (Mouse anti-HBeAg
mcAb), HBcAg vortex oscillating reactions 0.5-3h;
c)add bovine serum albumin(BSA) (BSA) respectively, lucifuge capping 0.5-2h;
d)the each product of centrifugal purification;
e)get each product precipitation and use the resuspended dispersion of PBS damping fluid respectively, obtain Mouse anti-HBsAg
mcAb-CdSe/ZnS QD
526, Mouse anti-HBeAg
mcAb-CdSe/ZnS QD
615, CdSe/ZnS QD
566-HBcAg each quantum point coupling thing solution.Preserve each quantum point coupling thing solution for 4 DEG C.
B. bar assembly preparation is tried:
a)sample pad 1: select cellulose membrane to make material, is cut into the film block with certain specification, and the phosphate buffer (PBS) put into by this film block containing 0.1%-10% BSA and 0.01%-10% Tween 20 soaks, and takes out, drying for standby.
b)red blood cell filter membrane 2: select red blood cell filter membrane to be cut into certain specification film block, drying for standby.
c)label pad 3: select glass fibre membrane to make material, is cut into the film block with certain specification, adds Mouse anti-HBsAg
mcAb-CdSe/ZnS QD
526, Mouse anti-HBeAg
mcAb-CdSe/ZnS QD
615, CdSe/ZnS QD
566the mixture solution of-HBcAg is on this film block, and desciccator diaphragm block is for subsequent use.
d)analyzing film 7: select nitrocellulose filter (NC film) to make material, be cut into the film block with certain specification, from film block base, make T by the lower potpourri from upper specking 0.5-10mg/ml rabbit AntiHBsAg antibody (Rabbit anti-HBsAg), the anti-HBeAg antibody of 0.5-10mg/ml rabbit (Rabbit anti-HBeAg) and 0.5-10mg/ml HBcAg respectively separated by a distance be with 4, specking 0.5-10mg/ml bis-anti-Quality Control thing sheep anti-mouse antibody makes C band 5, and desciccator diaphragm block is for subsequent use.
e)adsorptive pads 6: select the cellulose membrane with water sorption to be cut into the film block of certain specification, drying for standby.
C. bar assembly assembling is tried
The above-mentioned examination bar assembly prepared mutually overlaps in turn by sample pad 1, red blood cell filter membrane 2, label pad 3, analyzing film 7, adsorptive pads 6 and is pasted on plastics end liner 8, be cut into the examination bar of certain specification, load in plastic casing, load sealed storage in aluminium foil bag together with drying agent.
embodiment 3: the quantum dot immune chromatography examination bar of synchronous quantitatively joint inspection hepatitis B three is used for the preparation method of the synchronous quantitatively typical curve of joint inspection HBsAg, HBeAg, HBcAb
(a)getting HBsAg, HBeAg, HBcAb standard items uses phosphate buffer (PBS) to be made into the some parts of standard items series concentration in doubling dilution mode respectively.
(b)each standard concentration is dripped respectively and carries out detecting (that is: each standard concentration detects 10 times with detector under the same conditions with 10 quantum dot immune chromatography examination bars respectively) with detector under the same conditions on 10 quantum dot immune chromatography examination bars, read its T is with fluorescence intensity (OD respectively
t) be with fluorescence intensity (OD with C
c), obtain mean value and OD
t/ OD
cratio.
(c)x-axis is made, with OD with standard items series concentration
t/ OD
cratio makes Y-axis, obtains fluorescence intensity typical curve corresponding to concentration and sees Fig. 2.
Also standard items series concentration X-axis can be made, with OD
t/ (OD
t+ OD
c) ratio makes Y-axis, obtain fluorescence intensity typical curve corresponding to concentration, the present embodiment does not show.
embodiment 4: the quantum dot immune chromatography examination bar of synchronous quantitatively joint inspection hepatitis B three is used for the method for synchronous quantitatively joint inspection HBsAg, HBeAg, HBcAb
(a)typical curve data (these typical curve data also can adopt the storage mediums such as Quick Response Code, bar code or IC card chip to store, and the present embodiment does not show) are stored by RFID (RFID tag).
(b)storage has the RFID of typical curve data to be attached on examination bar, or is directly attached on the examination barrel of loading examination bar.
(c)the typical curve data of RFID storage are read and fluorescence intensity corresponding to the testing sample recorded in conjunction with detector and HBsAg, HBeAg, HBcAb concentration of obtaining in sample with the detector with signal testing function.
Examination bar of the present invention shown in Fig. 3 tries bar T when being used for synchronous quantitatively detection blood HBsAg, HBeAg, HBcAb and is with the fluorescence spectrum figure recorded.Wherein I is quantum dot (the CdSe/ZnS QD of HBsAg
526) fluorescence peak, II is quantum dot (the CdSe/ZnS QD of HBcAb
566) fluorescence peak, III is quantum dot (the CdSe/ZnS QD of HBeAg
615).
Special needs to be pointed out is: (1) embodiment of the present invention and accompanying drawing thereof are only in order to the present invention is described, those skilled in the art should not limit the scope of the invention with this; (2) the quantum dot immune chromatography examination bar of synchronous quantitatively joint inspection hepatitis B three of the present invention and method thereof naturally comprise the hepatitis B individual event that realized by identical examination bar structure, principle and method or many index detects; (3) the present invention can also have other to improve one's methods.Therefore, every other technical scheme adopting any equivalent replacement or equivalent transformation to be formed to examination bar of the present invention and method thereof, all drops in the protection domain of the claims in the present invention.
Claims (7)
1. the quantum dot immune chromatography examination bar of a synchronous quantitatively joint inspection hepatitis B three, comprise the sample pad (1) that overlap joint is in turn fixed on end liner (8), red blood cell filter membrane (2), label pad (3), analyzing film (7), adsorptive pads (6), analyzing film (7) has T band (4) and C band (5), be characterised in that: label pad (3) is coated with the mouse-anti HBsAg monoclonal antibody of different wave length quantum dot correspondence markings, the potpourri of mouse-anti HBeAg monoclonal antibody and HBcAg, T band (4) of analyzing film (7) is coated with rabbit AntiHBsAg antibody, the anti-HBeAg antibody of rabbit, the potpourri of HBcAg, C band (5) of analyzing film (7) is coated with Quality Control thing sheep anti-mouse antibody.
2. the quantum dot immune chromatography examination bar of synchronous quantitatively joint inspection hepatitis B three according to claim 1, is characterised in that: wherein said quantum dot comprises ZnS, CdS, HgS, ZnSe, CdSe, HgSe, CdTe, ZnTe, ZnO, PbSe, HgTe, CaAs, InP, InAs, InCaAs, CdS/ZnS, CdS/Ag
2s, CdS/PbS, CdS/Cd (0H)
2, CdS/HgS, CdS/HgS/CdS, ZnS/CdS, ZnS/CdS/ZnS, ZnS/HgS/ZnS/CdS, CdSe/CdS, CdSe/ZnS, CdSe/ZnSe, CdSe/CuSe, CdSe/HgTe, CdSe/HgSe, CdSe/HgSe/CdSe, CdTe/HgS, CdTe/HgTe, InAs/InP, InAs/CdSe, InAs/ZnSe, MgS, MgSe, MgTe, CaS, CaSe, CaTe, SrS, SrSe, SeTe, BaS, BaSe, BaTe, CdS:Mn, ZnS:Mn, CdS:Cu, ZnS:Cu, CdS:Tb, the combination of any one or any several nano particle in ZnS:Tb, and be core by any one quantum dot above-mentioned, silicon dioxide is the core-shell type nano-complex particle of shell.
3. the quantum dot immune chromatography examination bar of synchronous quantitatively joint inspection hepatitis B three according to claim 1, be characterised in that: described label pad (3) is glass fibre membrane, described analyzing film (7) is nitrocellulose filter, nylon membrane or cellulose nitrate/cellulose acetate hybrid films, and end liner (8) is polyester or plastic plate.
4. a preparation method for the quantum dot immune chromatography examination bar of synchronous quantitatively joint inspection hepatitis B three as claimed in claim 1, it is characterized in that, this examination bar is preparation method comprise the steps:
A. quantum point coupling antibody/antigen:
a)get different wave length quantum dot and adjust pH=6-9 with PBS damping fluid respectively, add EDC(l-(3-dimethyl aminopropyl)-3-ethyl carbon diamine hydrochloride) and NHS(N-hydroxy succinimide) room temperature activation 10-60min;
b)correspondence adds mouse-anti HBsAg monoclonal antibody, mouse-anti HBeAg monoclonal antibody and HBcAg vortex oscillating reactions 0.5-3h respectively;
c)correspondence adds BSA capping 0.5-2h respectively;
d)centrifugal purification product;
e)get the resuspended dispersion of precipitation PBS damping fluid, 4 DEG C of preservations;
B. bar assembly preparation is tried:
a)sample pad (1): select cellulose membrane to be cut into the film block of certain specification, soaked by the PBS that this film block is put into containing 0.1%-10% BSA and 0.01%-10% Tween 20, takes out, drying for standby;
b)red blood cell filter membrane (2): select red blood cell filter membrane to be cut into certain specification film block, drying for standby;
c)label pad (3): select glass fibre membrane to be cut into certain specification film block, add the mixture solution of the mouse-anti HBsAg monoclonal antibody by different wave length quantum dot correspondence markings, mouse-anti HBeAg monoclonal antibody and HBcAg on this film block, desciccator diaphragm block is for subsequent use;
d)analyzing film (7): select cellulose membrane to be cut into the film block of certain specification, from film block base, make T by the lower potpourri from the upper 0.5-10mg/ml of specking respectively rabbit AntiHBsAg antibody separated by a distance, the anti-HBeAg antibody of 0.5-10mg/ml rabbit, 0.5-10mg/ml HBcAg be with (4), specking 0.5-10mg/ml Quality Control thing sheep anti-mouse antibody makes C band (5), and desciccator diaphragm block is for subsequent use;
e)adsorptive pads (6): select the cellulose membrane with water sorption to be cut into the film block of certain specification, drying for standby;
C. bar assembly assembling is tried:
The above-mentioned examination bar assembly prepared overlaps in turn by sample pad (1), red blood cell filter membrane (2), label pad (3), analyzing film (7), adsorptive pads (6) and is pasted on end liner (8), be cut into the examination bar of certain specification, load in plastic casing, load sealed storage in aluminium foil bag together with drying agent.
5. the quantum dot immune chromatography examination bar of a synchronous quantitatively joint inspection hepatitis B three as claimed in claim 1 is used for the preparation method of the synchronous quantitatively typical curve of joint inspection HBsAg, HBeAg, HBcAb, it is characterized in that, this typical curve is preparation method comprise the steps:
(a)prepare each standard items series concentration of HBsAg, HBeAg, HBcAb respectively, and be added drop-wise on the quantum dot immune chromatography examination bar of described synchronous quantitatively joint inspection hepatitis B three;
(b)fluorescence intensity (the OD of T band is read respectively with detector
t) and C band fluorescence intensity (OD
c), calculate and obtain OD
t/ OD
cratio or OD
t/ (OD
t+ OD
c) ratio;
(c)x-axis is made, OD with standard items series concentration
t/ OD
cratio makes Y-axis, or makes X-axis with standard items series concentration, OD
t/ (OD
t+ OD
c) ratio makes Y-axis, obtains fluorescence intensity typical curve corresponding to concentration.
6. the quantum dot immune chromatography examination bar of synchronous quantitatively joint inspection hepatitis B three as claimed in claim 1 is used for a method of synchronous quantitatively joint inspection HBsAg, HBeAg, HBcAb, and it is characterized in that, this quantivative approach comprises the steps:
(a)by electronic tag storage typical curve data;
(b)storage has the electronic tag of typical curve data to be arranged on described examination bar, or is arranged on the examination barrel of loading examination bar;
(c)with there are the typical curve data of detector reading electronic labels storage of signal testing function and fluorescence intensity corresponding to the testing sample recorded in conjunction with detector and HBsAg, HBeAg, HBcAb concentration of obtaining in sample.
7. method according to claim 6, is characterized in that, wherein said electronic tag comprises RFID, Quick Response Code, bar code or the IC card chip with information storage function.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410544442.2A CN104297483A (en) | 2014-10-15 | 2014-10-15 | Quantum dot immunochromatography test strip for synchronous and quantitative joint inspection of HBsAg, HBeAg and HBcAb and method for synchronous and quantitative joint inspection of HBsAg, HBeAg and HBcAb |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410544442.2A CN104297483A (en) | 2014-10-15 | 2014-10-15 | Quantum dot immunochromatography test strip for synchronous and quantitative joint inspection of HBsAg, HBeAg and HBcAb and method for synchronous and quantitative joint inspection of HBsAg, HBeAg and HBcAb |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104297483A true CN104297483A (en) | 2015-01-21 |
Family
ID=52317300
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410544442.2A Pending CN104297483A (en) | 2014-10-15 | 2014-10-15 | Quantum dot immunochromatography test strip for synchronous and quantitative joint inspection of HBsAg, HBeAg and HBcAb and method for synchronous and quantitative joint inspection of HBsAg, HBeAg and HBcAb |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104297483A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113075411A (en) * | 2021-03-29 | 2021-07-06 | 重庆新赛亚生物科技有限公司 | Intercellular adhesion molecule-1 detection kit, preparation method and preparation device thereof |
| CN113150127A (en) * | 2021-05-10 | 2021-07-23 | 北京保图生物技术有限公司 | Nucleic acid antibody kit for rapidly detecting virus |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101246171A (en) * | 2008-01-29 | 2008-08-20 | 马义才 | Portable hepatitis B fast joint inspection device |
| CN101290319A (en) * | 2007-04-16 | 2008-10-22 | 北京出入境检验检疫局检验检疫技术中心 | H5N1 type highly pathogenic avian influenza nanometer quantum point detection method |
| CN101514987A (en) * | 2008-07-14 | 2009-08-26 | 马义才 | System for quantitative detection of quanta dot mark test bar and detection method thereof |
| CN103645311A (en) * | 2013-11-13 | 2014-03-19 | 成都领御生物技术有限公司 | Test strip card |
| CN103675266A (en) * | 2013-11-14 | 2014-03-26 | 成都领御生物技术有限公司 | Test strip card based on quantum dot labeling |
-
2014
- 2014-10-15 CN CN201410544442.2A patent/CN104297483A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101290319A (en) * | 2007-04-16 | 2008-10-22 | 北京出入境检验检疫局检验检疫技术中心 | H5N1 type highly pathogenic avian influenza nanometer quantum point detection method |
| CN101246171A (en) * | 2008-01-29 | 2008-08-20 | 马义才 | Portable hepatitis B fast joint inspection device |
| CN101514987A (en) * | 2008-07-14 | 2009-08-26 | 马义才 | System for quantitative detection of quanta dot mark test bar and detection method thereof |
| CN103645311A (en) * | 2013-11-13 | 2014-03-19 | 成都领御生物技术有限公司 | Test strip card |
| CN103675266A (en) * | 2013-11-14 | 2014-03-26 | 成都领御生物技术有限公司 | Test strip card based on quantum dot labeling |
Non-Patent Citations (1)
| Title |
|---|
| 马义才: "量子点免疫层析试条定量检测系统的设计", 《成都工业学院学报》, vol. 16, no. 4, 31 December 2013 (2013-12-31), pages 10 - 13 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113075411A (en) * | 2021-03-29 | 2021-07-06 | 重庆新赛亚生物科技有限公司 | Intercellular adhesion molecule-1 detection kit, preparation method and preparation device thereof |
| CN113150127A (en) * | 2021-05-10 | 2021-07-23 | 北京保图生物技术有限公司 | Nucleic acid antibody kit for rapidly detecting virus |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106872420B (en) | Kit and method for time-resolved fluorescence quantitative detection of microalbuminuria | |
| CN102520192B (en) | Fluorescence immunochromatographic assay and kit for quantitative detection of troponin I/creatine kinase isoenzyme/myohemoglobin | |
| CN102539785B (en) | Fluorescent immunochromatography method for whole quantitative detection of C-reactive protein and reagent kit thereof | |
| CN102520194B (en) | Fluorescence immunochromatographic assay kit for quantitatively detecting heart fatty acid binding protein | |
| CN100473989C (en) | Membrane-based detection utilizing time-resolved fluorescence | |
| US11371987B2 (en) | Method of amplifying detection light using light-reflecting material, in immunochromatography | |
| CN108080042A (en) | Micro-fluidic chip of binding time resolved fluorometric technology and its preparation method and application | |
| CN104267182A (en) | Quantum dot immunochromatographic strip for synchronously quantifying multiple tumor markers and method of quantum dot immunochromatographic strip | |
| CN103675266A (en) | Test strip card based on quantum dot labeling | |
| US20140170674A1 (en) | Membraine-Based Assay Devices Utilizing Time-Resolved Up-Converting Luminescence | |
| CN102565422B (en) | Fluorescence immunochromatographic assay and kit for quantitatively detecting cardiac troponin T | |
| CN203759009U (en) | Quantum dot labeled test strip card | |
| CN103630681A (en) | Test strip card based on quantum dot labeling | |
| CN103630683A (en) | Test strip card | |
| CN203759015U (en) | Test strip card | |
| CN102520193B (en) | Fluorescence immunochromatographic assay and kit for quantitative detection of human cardiac troponin I (cTnI) | |
| CN103630682A (en) | Test strip card based on quantum dot labeling | |
| KR101718485B1 (en) | Device for Detecting Colored Reaction or Fluorescence Reaction of Immunochromatography | |
| CN203759014U (en) | Test strip card | |
| CN104198702A (en) | Quantum-dot marking test strip capable of quantitatively determining multiple indexes of blood infectious diseases and preparation method and quantitative determination method thereof | |
| CN103630685A (en) | Test strip card | |
| CN103675267A (en) | Test strip card based on quantum dot labeling | |
| CN203759011U (en) | Quantum dot labeled test strip card | |
| CN103645312A (en) | Test strip card | |
| CN103630684A (en) | Test strip card |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150121 |