CN109541225A - A kind of detection hepatitis B virus e antigen kit and preparation method thereof - Google Patents
A kind of detection hepatitis B virus e antigen kit and preparation method thereof Download PDFInfo
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- CN109541225A CN109541225A CN201811329535.8A CN201811329535A CN109541225A CN 109541225 A CN109541225 A CN 109541225A CN 201811329535 A CN201811329535 A CN 201811329535A CN 109541225 A CN109541225 A CN 109541225A
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- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
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- 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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
Abstract
The invention discloses a kind of detection hepatitis B virus e antigen kits and preparation method thereof, are grouped as by following group: being coated with the magnetic microsphere, HBeAg calibration object, HBeAg antibody marker, analysis buffer, cleaning solution, enhancement solution and RFID card in conjunction with lanthanide series of HBeAg antibody.This method is based on magnetic microsphere and combines with Timed-resolved fluoroimmunoassay, overcome the physical sorption reaction time of ELISA Plate longer, the slower drawback of testing result, the reaction time is greatly shortened, while also having that time resolution detection technique accuracy height, high sensitivity, high specificity, the range of linearity is wide, detection is stable and convenient advantage.Magnetic microsphere is coated with corresponding antigen or antibody, because of the characteristic of magnetic microsphere solid, greatly increase the contact surface area of immune response, to greatly shorten detection time, and the present invention advanced optimizes traditional two-step method detection, detection HBeAg only needs one-step method to detect, and result can be detected in 30 minutes.
Description
Technical field
The present invention relates to a kind of detection hepatitis B virus e antigen kits and preparation method thereof, and in particular to Yi Zhongji
The preparation method of detection hepatitis B virus e antigen kit is combined with Timed-resolved fluoroimmunoassay in magnetic microsphere.
Background technique
Hepatitis B (abbreviation hepatitis B) is to lead to liver property inflammation and liver function energy loss caused by hepatitis type B virus (HBV)
A kind of harmful communicable disease.HBeAg is a kind of soluble protein, is generally detected in HBsAg positive serum.Acute HBV infection
When HBeAg time of occurrence be slightly later than HBsAg.The presence of HBeAg indicates that patient is in the high infection low response phase.HBeAg disappears
Instead anti-HBe afterwards.
The project of hepatitis B Serological testing at present generally comprises HBsAg, Anti-HBs, HBeAg, Anti-HBe, Anti-
HBc, that is, " Hepatitis B virus " being commonly called as.The detection method of existing mainstream HBeAg has enzyme linked immunological (ELISA), colloidal gold method, chemistry
Luminous (CLIA), electrochemical luminescence (ECL), time-resolved fluoroimmunoassay (TRFIA) etc..ELISA is as half-quantitative detection
Method is now most widely used detection method, but its sensitivity, linear extent range are not up to higher level, it is difficult to adapt to city
The demand of field development.Colloidal gold method and ELISA have the shortcomings that identical.Though and CLIA and ECL high sensitivity but presence detection
Equipment manufacturing cost is high, and corresponding marker research and development threshold is high, and the country can be under one's control in short-term, equally also limits
Domestic popularization.And time-resolved fluoroimmunoassay (TRFIA) sensitivity can reach the consistent level of CLIA, not only detection is set
It is standby cheap, and labelling technique is quite mature at home, testing cost is low simultaneous.And traditional time resolution detection, still
Based on the physical absorption of blank ELISA Plate, time-consuming for detection process.Shorten the reaction time, reduce testing cost, improves detection
Sensitivity and the range of linearity, will be with good market prospects.
Summary of the invention
Based on this, it is an object of the invention to overcome defect in the prior art and the relevant technologies testing cost is high lacks
Point provides the preparation method of a kind of detection hepatitis B virus e antigen kit and the kit.
To achieve the above object, the present invention adopts the following technical scheme that:
A kind of detection hepatitis B virus e antigen kit, is grouped as by following group: being coated with the magnetism of HBeAg antibody
Microballoon, HBeAg calibration object, the HBeAg antibody marker in conjunction with lanthanide series, analysis buffer, concentration washing lotion (cleaning solution)
With enhancement solution, RFID card.
Preferably, the magnetic microsphere in the magnetic microsphere for being coated with HBeAg antibody is by micron-sized Fe2O3Or
Fe3O4Magnetic particle and high-molecular organic material carry out it is compound, formed have superparamagnetism, can be with immunising antigen or anti-
The micron-sized microballoon that body combines, that is, be commonly referred to as " magnetic bead ".It is 0.1~5 μm that magnetic microsphere, which should be able to meet diameter, magnetic microsphere
Various active functional group, including but not limited to hydroxyl (- OH), amino (- NH can be had by surface modification2), carboxyl (-
COOH)。
Preferably, the HBeAg antibody in the magnetic microsphere for being coated with HBeAg antibody can be that monoclonal is anti-
Body is also possible to polyclonal antibody.
Preferably, the magnetic microsphere for being coated with HBeAg antibody is prepared using following steps:
By HBeAg antibody after 2~8 DEG C of superspeed refrigerated centrifuges are by buffer system replacement Treatment, after cleaning, activation
Magnetic microsphere mixing and constant-temperature incubation clean magnetic bead and abandon supernatant, then closed, cleaned with magnetic bead confining liquid after incubation
Magnetic bead abandons supernatant again, and the HBeAg antibody being coated with is saved in liquid in magnetic bead and is uprightly saved with 2~8 DEG C of refrigerators.
It is 10 μ g that every milligram of magnetic microsphere of EDC and Sulfo-NHS, which is preferably loaded quality, in the activation of the magnetic microsphere
~1000 μ g.Activation method includes but is not limited to EDC, Sulfo-NHS one such and two kinds.
Preferably, in order to reach quantitative purpose, the HBeAg calibration object can with preferred concentration for 0.6NCU/mL~
The calibration object solution and preferred concentration of 10NCU/mL is the calibration object solution of 100NCU/mL~500NCU/mL.
Preferably, the antibody in the HBeAg antibody marker in conjunction with lanthanide series is either monoclonal antibody also may be used
It how anti-is.Lanthanide series therein is to be completed in conjunction with HBeAg antibody by intermediate chelating agent, and lanthanide series includes but is not limited to
Europium (EU), samarium (Sm), chelating agent include but is not limited to isothiocyanic acid phenyl-EDTA, isothiocyanic acid benzyl-DTTA, the different sulphur of P-
Cyanobenzyls-DTTA, diethylene triamine pentaacetic acid aminophenyl-EDTA.
The present invention also provides a kind of methods using mentioned reagent box detection HBeAg, and described method includes following steps:
(1) marker is diluted to working solution with analysis buffer；
(2) working solution is diluted to by washing lotion is concentrated with purified water；
(3) magnetic microsphere of the working concentration diluted is added in reaction cup；
(4) sample to be examined or calibration object are added in above-mentioned reaction cup；
(5) the marker working solution diluted in (1) is added in reaction cup；
(6) reaction cup is incubated at room temperature；
(7) magnetic microsphere after using the cleaning liquid in (2) to add magnetic in washing reaction cup；
(8) after washing, degaussing is added enhancement solution and is incubated for；
(9) after being incubated for, the fluorescent collecting for carrying out corresponding wavelength is detected and is analyzed.
Preferably, for the present invention in order to further reduce manual steps, self-produced SmartTRF grinds certainly in cooperation company
The relevant parameter of detection method, operating procedure are all copied to RFID card by complete series Immunofluorescence test equipment
In.In actual mechanical process, it is only necessary to RFID card is adapted to above-mentioned Immunofluorescence test equipment can be automatically finished it is above-mentioned
The operating procedure of experiment.RFID (Radio Frequency Identification) technology, also known as radio frequency identification, is one
The kind communication technology can be identified specific objective by radio signals and read and write related data, without identifying system and specific mesh
Mechanical or optical contact is established between mark.
The present invention also provides a kind of preparation methods for detecting HBeAg kit, and described method includes following steps:
(1) preparation is coated with the magnetic microsphere of HBeAg antibody；
(2) HBeAg antibody marker of the preparation in conjunction with lanthanide series；
(3) calibration object is prepared；
(4) analysis buffer, concentration washing lotion and enhancement solution are prepared；
(5) each component is dispensed respectively into corresponding storage container；
(6) RFID card duplicate copy；
(7) coding, labelling；
(8) it is assembled into finished product kit.
Compared with prior art, the beneficial effects of the invention are that:
This method is based on magnetic microsphere and combines with Timed-resolved fluoroimmunoassay, that is, overcomes the physical absorption of ELISA Plate anti-
Longer between seasonable, the slower drawback of testing result greatly shortens the reaction time, while it is accurate also to have time resolution detection technique
Property height, high sensitivity, high specificity, the range of linearity is wide, detection is stable and convenient advantage.Magnetic microsphere is coated with corresponding antigen
Or antibody greatly increases the contact surface area of immune response because of the characteristic of magnetic microsphere solid, thus when greatly shortening detection
Between, and the present invention advanced optimizes traditional two-step method detection, and detection HBeAg only needs one-step method to detect, and 30 points
Result can be detected in clock.In addition, the frame limitation of traditional ELISA Plate is no longer limited by due to the fluid behaviour of magnetic microsphere,
Detection can be completed in arbitrary reaction cup and small test tube, at the same can reduce detecting instrument equipment volume size and equipment at
Originally to meet the full-automatic detection demand in two, three line cities, also big flux can be carried out such as ELISA Plate, the reflective detection of chemistry
Detection meets the full-automatic detection demand in a line city.
Detailed description of the invention
Fig. 1 is to be used to store the magnetic microsphere and group of the lanthanides for being coated with HBeAg antibody using detection kit of the present invention
The schematic top plan view of the reagent strip of HBeAg antibody marker, analysis buffer, cleaning solution and enhancement solution that element combines.
Fig. 2 is to be used to store the magnetic microsphere and group of the lanthanides for being coated with HBeAg antibody using detection kit of the present invention
The stereochemical structure mark intention of the reagent strip of HBeAg antibody marker, analysis buffer, cleaning solution and enhancement solution that element combines is shown
It is intended to.
Specific embodiment
The present invention is further illustrated with attached drawing with reference to embodiments, and it is special that technology of the invention is better described
Point, technical solution.Following embodiment does not cause any restrictions to the present invention.HBeAg antigen, antibody come in following embodiment
Derived from Wuhan AudioCodes Bo Tai Biotechnology Co., Ltd；Magnetic microsphere derives from GE company；Europium label is purchased from Finland Wallac public affairs
Department；Hepatitis B virus e antigen National reference is provided by National Institute for Food and Drugs Control；Contrast agents box is Abbott Laboratories' second
Hepatitis virus e Antigen quantitative determination kit (chemiluminescence particulate immunodetection).
Embodiment 1
A kind of detection hepatitis B virus e antigen kit, the kit include: the magnetism for being coated with HBeAg antibody
Microballoon, HBeAg calibration object, the HBeAg antibody marker of europium label, analysis buffer, concentration washing lotion and enhancement solution, RFID card.
The present invention also provides the preparation methods of above-mentioned detection hepatitis B virus e antigen kit, and the method includes such as
Lower step:
(1) preparation is coated with the magnetic microsphere of HBeAg antibody: will through 2~8 DEG C of superspeed refrigerated centrifuges by HBeAg antibody
After buffer system replacement Treatment, mixed simultaneously constant-temperature incubation 1~3 hour with 1 μm of carboxyl magnetic microsphere of diameter after cleaning, activation,
Magnetic bead is cleaned using magnetic bead cleaning solution after incubation and abandons supernatant, is then closed with magnetic bead confining liquid, and cleaning magnetic bead is again
The HBeAg antibody magnetic microsphere being coated with is saved in liquid in magnetic bead and is uprightly saved with 2~8 DEG C of refrigerators by secondary discarding supernatant.Again
HBeAg antibody magnetic microsphere will be coated with and be diluted to working solution concentration with magnetic bead preservation liquid, be distributed into 10mL/ bottles.Preferably, magnetic
Property microballoon and the coated mass ratio of HBeAg antibody be one of 10:1,20:1,30:1,40:1；Preferably, the displacement
Buffer and magnetic bead cleaning solution are the MES buffers of 0.05~0.5M PH, 5.8~PH 7.0；Preferably, the work of magnetic microsphere
It is 10 μ of μ g~1000 g that every milligram of magnetic microsphere of EDC and Sulfo-NHS, which is preferably loaded quality, in change；Preferably, magnetic microsphere
Confining liquid and save liquid be 0.1~0.5M PH 7.0~8.5 containing 0.1%~8%BSA Tris-HCl buffer；
(2) prepare europium label HBeAg antibody marker: by HBeAg antibody be placed in molecular cut off be 10000 ultrafiltration from
In heart pipe, 10000rpm is centrifuged 7~10min, discards filtrate.Add 9.6 carbonate buffer solution 10000rpm of 0.05M PH from
7~10min of the heart 2~3 times repeatedly, centrifuge tube filter membrane reversion 3000rpm is centrifuged 6min, collects 200 μ L solution being finally concentrated.
And by it and in advance with the solvent DTTA-EU of carbonate buffer solution3+Mixing, HBeAg antibody and europium mass ratio are 1:1,2~8 DEG C
Oscillation mixes 48 ± 2 hours.The Sephadex TM G-75 that label solution is balanced through 0.05M PH 7.8Tris-Hcl buffer
Gel columnChromatographic purifying monitors in A280 and collects first peak.The HBeAg antibody mark that the europium being collected into is marked
Remember that object 0.2%BSA, 0.05M PH 7.8Tris-Hcl buffer are diluted to 1/20 times of optium concentration, packing to 1.0mL/
Bottle.
(3) it prepares HBeAg calibration object: using 2%BSA, 0.5%Tween-20,0.02%Proclin300, pH 7.5
0.02M Tris-HCl buffer is by HBeAg antigen diluent to lower concentration values point 2.5NCU/mL, high concentration value point 160NCU/mL.
(4) it prepares analysis buffer: containing Tween-20, Proclin300, EDTA, BSA, Tris-HCl buffer, dividing
It is filled to 30~40mL/ bottles.
(5) preparation concentration washing lotion: containing Tween-20, Proclin300, Tris-Hcl buffer, dispense to 30~
40mL/ bottles.
(6) enhancement solution is prepared: containing sodium acetate, β-NTA, TOPO, glacial acetic acid, dehydrated alcohol, Triton X-100, packing
To 30~40mL/ bottles.
(7) it prepares RFID card: blank RFID card is subjected to relative parameters setting by detection method in the present embodiment of the present invention；
(8) coding, labelling assemble kit.
The present invention also provides the detection method of above-mentioned detection hepatitis B virus e antigen kit, the method tools
Gymnastics is made as follows:
(1) reagent prepares
1. kit restores in being placed at room temperature for room temperature；
2. the HBeAg antibody marker of europium label is diluted to working solution concentration using analysis buffer, mix stand-by；
3. cleaning solution: purified water is added by 1:25 in concentration washing lotion and is diluted to work cleaning solution；
4. upright light rolling is coated with the magnetic microsphere of HBeAg antibody before experiment, mix stand-by；
(2) experimental implementation
1. the magnetic microsphere for drawing 50 μ L mixing is added in reaction cup；
2. sample to be tested or 100 μ L of calibration object are added into each reaction cup；
3. the 100 μ L of HBeAg antibody marker working solution diluted is added into each reaction cup again；
4. room temperature blending incubation 30 minutes；
5. after being incubated for, being cleaned 4 times using cleaning solution；
6. 100 μ L of enhancement solution is added into each reaction cup, room temperature blending incubation 3 minutes；Fluorescence is completed in 30 minutes
It acquires and carries out data analysis.
In the present embodiment, actual laboratory operating procedures and relevant parameter are copied in matched RFID card in advance, real
It tests after operating process only needs to be ready to by reagent preparation process, RFID card is adapted to fully-automatic equipment can be completed from information
Read, be loaded onto the overall process of detection.
Embodiment 2
A kind of detection hepatitis B virus e antigen kit, it is essentially identical with detection kit described in embodiment 1, no
It is same to be:
(1) the detection hepatitis B virus e antigen kit component only includes: reagent strip, HBeAg calibration object, RFID
Card.
(2) in the present embodiment, reagent strip is by being coated with the magnetic microsphere of HBeAg antibody, europium in the detection kit
In HBeAg antibody marker, analysis buffer, cleaning solution and the enhancement solution of label, packing to the corresponding hole of reagent strip after sealer
It forms.Wherein cleaning solution is that concentration is washed in embodiment 1 plus purified water dilutes 25 times and forms；Remaining each component and phase in embodiment 1
Together.
In the present embodiment, just as shown in Figure 1 and Figure 2, each hole bit function of reagent strip is described as follows in the detection kit:
Reagent strip is from left to right arranged successively, and title is followed successively by the 1st~13 hole.1st, 2 holes are instrument connection, the 3rd, 4
Hole be fluorescent marker hole, the 5th hole be analysis buffer hole, the 6th, 7 holes be cleaning fluid apertures, the 8th, 9 be Sample Dilution fluid apertures, the 12nd
To enhance fluid apertures, the 10th, 11,13 be preparation hole.1st and 2 holes are the reacting hole storing magnetic microsphere and being immunoreacted, most
It is 800 μ L that liquid volume can be stored greatly；3rd, 4 holes can be disassembled into from entire reagent strip and be independent component, convenient for glimmering
Signal object carries out packing storage.3rd, 4,5 holes can store maximum liquid volume be 400 μ L；6th, 7 holes can store maximum
Liquid volume is 3000 μ L；It is 400 μ L that 8th~12 hole, which can store maximum liquid volume,；13rd hole can store maximum liquid
Volume is 600 μ L.
In the present embodiment, the reagent strip in the detection kit is made after carrying out sealer as follows: by 300 μ L
It is coated with HBeAg antibody magnetic microsphere, 50 μ L europiums label HBeAg antibody marker, 200 μ L analysis buffers, 3000 μ L cleaning
Liquid, 200 μ L enhancement solutions are dispensed respectively to the 1st of reagent strip the, 3,5,6,12 holes, and the reagent strip is made after sealer.
The present invention also provides the preparation methods of above-mentioned detection hepatitis B virus e antigen kit, are obtained in the above describe manner
After reagent strip, kit is constituted with HBeAg calibration object, RFID card.In addition to each component packing mode and storage container are different
Outside sample, remaining is in the same manner as in Example 1.
The present invention also provides the detection methods of above-mentioned detection hepatitis B virus e antigen kit, it is only necessary to by above-mentioned examination
Agent item is inserted into the reagent clamp bar slot of SmartTRF serial equipment after gently shaking mixing, and equipment reads RFID card relevant information can be complete
It is automatically performed detection process.The related information parameters and detecting step of RFID card are in the same manner as in Example 1.
Embodiment 3
The performance evaluation of detection hepatitis B virus e antigen kit of the present invention:
By the kit and detection method prepared in embodiment, second of the detection buying from National Institute for Food and Drugs Control
Hepatitis virus e antigen National reference, and collect facing from Abbott Laboratories, hospital Architect i2000 detection HBeAg
Bed 300, sample.
Embodiment 1, embodiment 2 detect hepatitis B virus e antigen National reference, as a result as follows:
(1) negative match-rate: 15 parts of negative National reference N1~N15, embodiment 1, embodiment 2 detect coincidence rate
(-/-) it is 15/15；
(2) positive coincidence rate: 10 parts of positive National reference P1~P10, embodiment 1, embodiment 2 detect coincidence rate (+/
+) it is 10/10；
(3) accuracy: accuracy reference material repeats to detect 10 times, and embodiment 1, the detection of embodiment 2 accuracy C.V≤
15%；
(4) limit of identification: detection sensitivity reference material, embodiment 1, the detection of embodiment 2 meet≤1IU/mL.
Embodiment 1, embodiment 2 detect 300 clinical samples of Abbott Laboratories HBeAg, as a result as follows:
The comparison of 1 embodiment of table, 1 clinical sample
Detection kit described in the embodiment of the present invention 1 detects hepatitis B e antigen and contrast agents negative match-rate
100%, positive coincidence rate 100%.
What the concentration washing lotion (cleaning solution) referred in embodiment 1 is that the high concentration to be diluted to working solution is cleaned
Liquid；What cleaning solution referred in example 2 is the working solution for not needing any processing.
It should be understood that the detection method and preparation method of detection HBeAg kit are big in order to meet in embodiment 1
Flux testing goal and invent, instrument and equipment bulky and cost is relatively high, for further satisfaction two, three line cities
Detection demand, correspondingly, we have further made some detection methods and kit preparation side on the basis of embodiment 1
Modification in method, as in embodiment 2.
It should be understood that difference of the present invention in example 2 with detection method and preparation method in embodiment 1 exists
In embodiment 2 is by the magnetic microsphere for being coated with HBeAg antibody in embodiment 1, the HBeAg antibody in conjunction with lanthanide series
Marker, analysis buffer, cleaning solution and enhancement solution are dispensed simultaneously into special reagent strip (as shown in Figure 1 and Figure 2), therefore real
The component for applying kit in example 2 only has reagent strip, RFID card, HBeAg calibration object.Wherein RFID card, HBeAg calibration object and implementation
It is consistent in example 1.
It should be understood that the detection method of the present invention in example 2 is after reagent strip is inserted into detection device, to use simultaneously
Corresponding RFID card is adapted to equipment, and equipment full automatic working step is consistent with embodiment 1；
It should be understood that the preparation method of the detection kit of the present invention in example 2 it is different from embodiment 1
In concentration washing lotion becomes cleaning solution in step (4), and step (5) becomes " dispensing each component respectively to the corresponding aperture of reagent strip
It is interior, sealer coding immediately after having dispensed ".
Claims (7)
1. a kind of detection hepatitis B virus e antigen kit, it is characterised in that be grouped as by following group: it is anti-to be coated with HBeAg
Magnetic microsphere, HBeAg calibration object, HBeAg antibody marker, analysis buffer, the cleaning solution, increasing in conjunction with lanthanide series of body
Strong liquid and RFID card.
2. detection hepatitis B virus e antigen kit as described in claim 1, which is characterized in that described is coated with
Magnetic microsphere in the magnetic microsphere of HBeAg antibody is by micron-sized Fe2O3Or Fe3O4Magnetic particle and organic polymer material
Material carries out compound, micron-sized microballoon that formation has superparamagnetism, can combining with immunising antigen or antibody.
3. detection hepatitis B virus e antigen kit as claimed in claim 2, which is characterized in that the magnetic microsphere
Diameter is 0.1~5 μm, and magnetic microsphere has various active functional group, including but not limited to hydroxyl, amino by surface modification
Or carboxyl.
4. detection hepatitis B virus e antigen kit as described in claim 1, which is characterized in that described is coated with
The magnetic microsphere of HBeAg antibody is prepared using following steps:
By HBeAg antibody after 2~8 DEG C of superspeed refrigerated centrifuges are by buffer system replacement Treatment, with the magnetism after cleaning, activation
Microballoon mixing and constant-temperature incubation clean magnetic bead and abandon supernatant, then closed with magnetic bead confining liquid, clean magnetic bead after incubation
Supernatant is abandoned again, and the HBeAg antibody being coated with is saved in liquid in magnetic bead and is uprightly saved with 2~8 DEG C of refrigerators.
5. detection hepatitis B virus e antigen kit as described in claim 1, which is characterized in that the HBeAg calibration
The calibration object that the calibration object solution and concentration that product concentration is 0.6NCU/mL~10NCU/mL are 100NCU/mL~500NCU/mL is molten
Liquid.
6. detection hepatitis B virus e antigen kit as described in claim 1, which is characterized in that described first with group of the lanthanides
The antibody in HBeAg antibody marker that element combines is monoclonal antibody or resists more；Lanthanide series is to pass through centre in conjunction with HBeAg antibody
Chelating agent is completed, and lanthanide series is europium or samarium, and chelating agent is isothiocyanic acid phenyl-EDTA, isothiocyanic acid benzyl-DTTA, P-
Isothiocyanatobenzyl-DTTA or diethylene triamine pentaacetic acid aminophenyl-EDTA.
7. the preparation method of detection hepatitis B virus e antigen kit described in claim 1, it is characterised in that including as follows
Step:
(1) preparation is coated with the magnetic microsphere of HBeAg antibody；
(2) HBeAg antibody marker of the preparation in conjunction with lanthanide series；
(3) calibration object is prepared；
(4) analysis buffer, concentration washing lotion and enhancement solution are prepared；
(5) each component is dispensed respectively into corresponding storage container；
(6) RFID card duplicate copy；
(7) coding, labelling；
(8) it is assembled into finished product kit.
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| CN110954692A (en) * | 2019-12-20 | 2020-04-03 | 蓝怡科技集团股份有限公司 | Reducing agent buffer solution and preparation method and application thereof |
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