CN101377503A - Hepatitis b virus preS2 antigen chemiluminescence immune analysis determination reagent kit and preparing method thereof - Google Patents
Hepatitis b virus preS2 antigen chemiluminescence immune analysis determination reagent kit and preparing method thereof Download PDFInfo
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Abstract
The invention discloses a detection kit for detecting hepatitis B virus pre-S2 antigens. The kit provided by the invention comprises calibrators, solid-phase vectors which are coated by avidin, biotinylation antibodies, anti-HBs markers, chemiluminescent substrates and concentrated washing solutions. The preparation of the kit comprises the following steps:1) preparing the calibrators with pure hepatitis B virus pre-S2 antigens, 2) coating the solid-phase vectors with the avidin, 3) biotinylating of hepatitis B virus pre-S2 antigen monoclonal antibodies, 4) marking the anti-HBs monoclonal antibodies with markers, 5) preparing the chemiluminescent substrates, 6) preparing the concentrated washing solutions, 7) packaging the calibrators, the markers, the chemiluminescent substrates and the washing solutions and 8) assembling finished products. The kit of the invention has the advantages of high sensitivity, strong specificity, fast detection speed, simple and convent operation, good repeatability and the like, can specially detect the content of the hepatitis B virus pre-S2 antigens in the human body after being affected by hepatitis B virus, and can be used as an auxiliary means for the diagnosis and the prognosis of hepatitis B.
Description
Technical field
The present invention relates to the immunoassay medical domain, particularly, the invention provides the indirect coating technique of a kind of biotin-avidin and measure hepatitis b virus preS 2 antigen (HBV Pre-S2Ag) kit and preparation method thereof in conjunction with chemiluminescence immunoassay technology.
Background technology
Hepatitis B (Hepatitis B, HB) be one of the whole world the widest popular viral infection disease, be by hepatitis type B virus (Hepatitis B virus, HBV) cause, with the liver inflammatory lesion serve as main and can cause multiple organ injury a kind of popular remote, propagate extensively, the serious infectious disease of harm.
Up to now, hepatitis b virus infected treatment does not have breakthrough progress as yet, and the health of masses problem that is caused by HBV also is far from being resolved.Therefore, hepatitis B has become the worldwide disease of serious threat human life health, to the control of hepatitis B become China healthy with communicable disease control in matter of utmost importance.
Hepatitis type B virus (HBV) genome total length only is 3200 bases, exists with part ring-type two strands.Wherein minus strand is complete loop chain, comprises 4 overlapped reading frames: preceding C/C district, S district, P district and X district.
The S district is by pre-s1 gene, anterior s2 gene and S genomic constitution, the 3 kinds of albumen of encoding respectively, are formed small-molecular weight master albumen (S albumen) by 226 amino acid at the large molecular weight protein (L albumen or pre-s1 protein) be made up of 409 amino acid of promptly encoding, the middle molecular weight protein of being made up of 285 amino acid (M albumen or pre-s2 protein).Preceding C/C gene, coding HBeAg and HBcAg.The P district is the longest, accounts for genome more than 75%, coding HBV DNA polymerase.X district (nucleotide 1,374～1,835) 154 the amino acid whose basic polypeptides of may having encoded, the breach of long-chain is positioned at this district.
Japanese scholar found to have a kind of material energy and polyerized human serum albumin (PHSA) combination with the blood clotting method in hepatitis B patient blood in 1992.Studies confirm that further this material is a kind of composition of hepatitis B shell, promptly preceding S2.Clinical data confirms that HBV enlivens when duplicating, and preceding S2 titre increases, otherwise is low titre or feminine gender.During acute hepatitis b, if preceding S2 disappears early, anti-preceding S2 occurs early, the then most recoveries from illness of patient, on the contrary then point out the possibility of chronicity.
The hepatitis b virus preS 2 antigen laboratory is detected and is mainly immunoassay, comprises methods such as radiommunoassay (RIA), enzyme-linked immuno assay (ELISA) and indirect agglutination method, and wherein ELISA is the main flow detection method in present clinical diagnosis market.Advantages such as that the ELISA detection method has is easy and simple to handle, easily popularize, but its enzyme labeling thing substrate is poisonous mostly or be carcinogenic substance, and instability, its sensitivity is difficult to satisfy the highly sensitive requirement in clinical market.Though the RIA detection method is highly sensitive, its term of validity is short, exist shortcoming such as radioactive waste pollution equally also to be difficult to satisfy the requirement in present clinical diagnosis market.In recent years, fast-developing chemiluminescence immune assay (Chemiluminescene CLIA) is a kind of cold ultramicron immunoassay new technology, it has inherited all advantages of RIA and ELISA, has overcome the two common shortcoming simultaneously, has highly sensitive, high specificity, advantages such as precision is good, and the range of linearity is wide, and method is stable, quick, becoming the substituent of radioimmunoassay and normal enzyme immunoassay, is that present immune quantitative is analyzed optimal method.
The application of Avidin-biotin system in ELISA has various ways, amplifies, wraps indirectly forms such as quilt comprising reaction signal.Reaction signal amplifies the Avidin reaction system that is biotinylated antigen or antibody and enzyme labeling, its advantage is to utilize the biotin-avidin enlarge-effect to improve detection sensitivity, shortcoming is that the isoelectric point of Avidin is higher, and solid phase carriers such as p-poly-phenyl ethene have higher non-special absorption and cause higher background.The present invention with the Avidin bag by solid phase carrier, wrap indirectly by biotinylated antibody by Avidin-biotin system, not only be convenient to amplify and produce, be easy to monitoring industrial processes, and shortcoming such as the loss of activity of having avoided the direct coated solid phase to exist is big, process optimization is loaded down with trivial details, the stability between having guaranteed simultaneously batch.
The objective of the invention is to put clinically at present in order to solve that the agent term of validity of being excused from an examination is short, radioactive waste contamination hazard and enzyme are excused from an examination deficiencies such as agent sensitivity is low, accuracy difference.Hepatitis b virus preS 2 antigen chemiluminescence of the present invention is measured kit, through clinical trial certificate, easy and simple to handle, highly sensitive, specificity is good, degree of conformity is high as a result to detect hepatitis B virus DNA with PCR, can be for the monitoring hepatitis B virus duplication provides assistance in diagnosis on protein level, for the diagnosis basis that provides of effective comprehensive therapeutic plan, prevention cirrhosis and liver cancer is provided as early as possible.
Summary of the invention
The present invention combines the indirect coating technique of biotin-avidin, chemiluminescence effectively with the immunoassay of hepatitis b virus preS 2 antigen, provides that a kind of highly sensitive, high specificity, detection speed are fast, easy and simple to handle, the hepatitis b virus preS 2 antigen detection kit of good reproducibility.
The purpose of this invention is to provide the kit of the indirect coating technique of a kind of biotin-avidin in conjunction with the chemiluminscence immunoassay hepatitis b virus preS 2.
Kit of the present invention is made up of the monoclonal antibody of the solid phase carrier of hepatitis b virus preS 2 antigen calibration object, Avidinization, biotinylated hepatitis b virus preS 2 antigen, anti--HBs labeling of monoclonal antibody thing, chemical luminous substrate and concentrated cleaning solution.
According to kit of the present invention, wherein, described solid phase carrier is microwell plate, plastic tube or magnetic-particle; Described label is alkaline phosphatase or chemiluminescence agent, and wherein said chemiluminescence agent is acridinium ester, luminol or different luminol; Described chemical luminous substrate is 1,2-two oxidative ethane analog derivatives, urea peroxide or hydrogen peroxide, wherein said 1,2-two oxidative ethane analog derivatives are (diamantane)-1,2-two oxidative ethanes, 3-(2 '-the spiral diamantane)-4-methoxyl-4-(3 " the phosphorus acyloxy) phenyl-1,2-two oxidative ethanes (AMPPD), CSPD or CDP-Star.
Another object of the present invention is for providing the preparation method of mentioned reagent box.
The Avidin bag by the method for solid phase carrier is: with 0.050M pH value is the Avidin coating buffer that 9.6 carbonate buffer solution is mixed with desired concn, and coating buffer is carried on the solid phase carrier, washs with physiological saline, after the sealing of 10% lowlenthal serum, the dehumidifier drying, aluminium foil bag is airtight, cryopreservation.Wherein, described bag is microwell plate, plastic tube or magnetic-particle by the solid phase carrier of Avidin.
With alkaline phosphatase or chemiluminescence agent and anti--HBs monoclonal antibody coupling, wherein said chemiluminescence agent is acridinium ester, luminol or different luminol with glutaraldehyde method.
With 1,2-two oxidative ethane analog derivatives, urea peroxide or hydrogen peroxide preparation chemical luminous substrate, wherein said 1,2-two oxidative ethane analog derivatives are (diamantane)-1,2-two oxidative ethanes, 3-(2 '-the spiral diamantane)-4-methoxyl-4-(3 " the phosphorus acyloxy) phenyl-1,2-two oxidative ethanes, CSPD or CDP-Star.
The prescription of concentrated cleaning solution is: the 0.2M pH7.4Tris-HCl damping fluid that contains 16% NaCl, 0.4% KCl.
Kit of the present invention adopts the indirect coating technique of Avidin-biotin to combine with chemiluminescence, avoided direct physical absorption to have shortcomings such as the immunocompetence loss is big, sterically hindered, accuracy difference, and simplified process optimization, be convenient to monitoring industrial processes.
Kit of the present invention utilizes Avidin-biotin system to wrap indirectly by pre-s2 antibody, form solid phase Avidin-bioid pre-s2 antibody-preS 2 antigen compound with preS 2 antigen reaction in the sample to be tested, anti--HBs reaction formation solid phase Avidin-biotinylation pre-s2 antibody-preS 2 antigen-alkali phosphatase enzyme mark with alkali phosphatase enzyme mark resists-the HBs sandwich complex again, the catalytic substrate chemiluminescence, quantitative test reflects the content of hepatitis b virus preS 2 in the serum, has avoided the chemiluminescence reaction plateau weak point of horseradish peroxidase-labeled thing catalysis, the unsettled shortcoming of chemical luminous substrate.
Kit of the present invention also can utilize the Avidin solid phase, form solid phase Avidin-bioid antibody-preS 2 antigen compound with preS 2 antigen reaction in biotinylation monoclonal antibody and the sample to be tested, anti--HBs reaction formation solid phase Avidin-biotinylated antibody-preS 2 antigen-acridinium ester mark with the acridinium ester mark resists-the HBs sandwich complex again, add chemical luminous substrate then, utilize Chemiluminescence Apparatus to detect relative luminous intensity, the content of hepatitis b virus preS 2 in the quantitative test reflection serum, and then embody the infection level of hepatitis type B virus.Sensitivity that agent exists is low, endogenous enzyme disturbs, the most of shortcoming such as poisonous or carcinogenic of substrate thereby avoided enzyme to be excused from an examination, and has advantages such as reaction velocity is fast, light signal is not subjected to influence of temperature change, cost is low, easy storage.Kit of the present invention can detect the content of the hepatitis b virus preS 2 antigen in the human body behind the hepatitis b virus infection very single-mindedly, the reflection hepatitis B virus infection with duplicate index, as the supplementary means of hepatitis diagnosis, prognosis.
Kit of the present invention is by the chromogenic substrate in the light signal replacement EIA enzyme immunoassay that detects the luminous substrate generation, thereby have a specificity equal with EIA enzyme immunoassay, and remolding sensitivity EIA enzyme immunoassay now improves greatly, and the diagnosis that can be hepatitis b virus preS 2 antigen provides more special, quick, reliable foundation.
Description of drawings
Fig. 1 is the calibration object linear graph in the prepared kit of embodiment 1.
Embodiment
In research process of the present invention, the inventor has at first carried out shaker test and Quality Identification to used starting material, comprises the activity of labelled antibody and biotinylated antibody, absorption property and accuracy, the chemiluminescence intensity etc. of Avidin solid phase carrier.Then Avidin solid-phase coating system has been carried out systematic research, contrasted different bag quilts, sealing buffer system, selected optimal bag and be cushioned liquid and confining liquid, and select best bag by concentration according to the square formation titration.The inventor by repeatedly comparative study biotin coupling antibody technology and the marking process of alkaline phosphatase, filter out the labeling method of high yield, low cost, reliable in quality, easy easy operating, and the dilution of biotin coupling antibody and enzyme labeling thing carried out deep system optimization, prepared and satisfied the label dilution that clinical practice is stablized needs.
One, the preparation of enzyme labelled antibody preparation and biotin coupling antibody
1, the monoclonal anti body and function glutaraldehyde method of hepatitis b virus preS 2 antigen and alkaline phosphatase coupling are fully dialysed to PBS, add equal-volume glycerine, preserve standby below-20 ℃.
2, the Monoclonal Antibody of biotin coupling hepatitis b virus preS 2 antigen
(1) with conventional method biotin (BNHS) is dissolved in N, dinethylformamide (DMF) is made into 1mg/mL;
(2) with 0.1mol/L pH value be 9.0 NaHCO
3With the monoclonal antibody dilution of the hepatitis b virus preS 2 antigen of purifying is 1～2mg/mL;
(3) be to mix about 1:8 or weight ratio 1:7 by BNHS with the antibody volumetric ratio, react 2～4h under the stirring at room;
(4) dialysis of packing into is 7.2 PBS to 0.05mol/L pH value, 4 ℃ of dialysed overnight, and bond adds equal-volume glycerine, packing in a small amount, preservation is standby below-20 ℃.
3, biotinylation monoclonal antibody, enzyme mark anti--preparation of HBs monoclonal antibody dilution
Tris 12.12g
Lowlenthal serum 100mL
Proclin?300 1mL
Distilled water 1000mL
4, biotinylation monoclonal antibody and enzyme labeling anti--preparation of HBs monoclonal antibody working fluid
With the biotinylation monoclonal antibody, enzyme labeling of preparation anti--the HBs monoclonal antibody is diluted to variable concentrations respectively with dilution, uses the chessboard titrimetry to select best dilutability for being respectively 1:4800,1:20000.
Two, the preparation of hepatitis b virus preS 2 antigen calibration object
With the pure product preparation of hepatitis b virus preS 2 antigen, totally 6 bottles of packing 0,1,5,10,50,100ng/mL.
Three, the solid phase microwell plate of Avidinization preparation
(1) bag quilt
The coating buffer compound method:
Natrium carbonicum calcinatum 0.795g
Sodium bicarbonate 1.47g
Distilled water 500mL
Behind the dissolving mixing, adjust pH to 9.6, add an amount of Avidin mixing, add then in each hole of microwell plate, every hole 130 μ L place 24h for 4 ℃.
(2) washing: it is inferior to give a baby a bath on the third day after its birth with physiological saline.
(3) sealing
The confining liquid compound method:
NaH
2PO
4·2H
2O 0.2g
Na
2HPO
4·12H
2O 2.9g
Lowlenthal serum 100mL
Proclin?300 1mL
Distilled water is settled to 1000mL
The mentioned reagent weighing is put into clean container well, add the distilled water constant volume, the dissolving mixing, measuring the pH value is 7.0.
Every hole adds confining liquid 120 μ L respectively, and room temperature was placed 4 hours.Get rid of confining liquid, on thieving paper, pat dry.Room temperature removal moisture drying 24 hours.Carry out vacuum sealing bag immediately.Place behind the envelope and checked not have gas leakage in 15 minutes, if there is gas leakage to need envelope again.2～8 ℃ of preservations of labeling postposition.
Four, chemical luminous substrate liquid
The compound method of the chemical luminous substrate liquid of alkaline phosphatase used in the present invention (ALP):
Tris 24g
HCl 15mL
NaCl 160g
KCl 4g
Proclin?300 1mL
AMPPD 200mL
Distilled water is fixed molten to 1000mL
Five, concentrated cleaning solution
Na
2HPO
4·12H
2O 58g
NaH
2PO
4·2H
2O 2g
NaCl 160g
Tween-20 1mL
Proclin?300 1mL
Distilled water is fixed molten to 1000mL
Dilute 20 times with distilled water during use.
Six, semi-manufacture and finished product are formed
The packing of above-mentioned steps products obtained therefrom is semi-manufacture.After extracting three parts of process specificitys, accuracy, sensitivity and stable assay approvals at random out, be assembled into hepatitis b virus preS 2 antigen and measure kit (chemoluminescence method).After sampling observation, behind the finished product kit assay approval, go into product library.
The present inventor has carried out systematic research to the reaction pattern and the reaction conditions of kit, finally determined double antibody sandwich method two-step reaction pattern, and the influence of test findings is tested with regard to the different reaction time, determine the optimal reaction time.By the influence test to measured value shows to the mensuration of luminous duration of chemical luminous substrate liquid and different fluorescent lifetime: be measured as the best between 30～60 minutes after adding chemical luminous substrate liquid, its result is also comparatively accurate.
Embodiment 2 preparations hepatitis b virus preS 2 antigen chemiluminescence immune analysis determination kit of the present invention
As outside the solid phase carrier, all the other all prepare the hepatitis b virus preS 2 antigen chemiluminescence immune analysis determination kit with the method identical with embodiment 1 divided by plastic tube.
Embodiment 3 preparations hepatitis b virus preS 2 antigen chemiluminescence immune analysis determination kit of the present invention
, Avidin is coupled to outside the magnetic-particle surface with the EDC method as solid phase carrier divided by magnetic-particle, all the other all prepare the hepatitis b virus preS 2 antigen chemiluminescence immune analysis determination kit with the method identical with embodiment 1.
Embodiment 4-6 prepares hepatitis b virus preS 2 antigen chemiluminescence immune analysis determination kit of the present invention
Divided by glutaraldehyde method with acridinium ester, luminol or different luminol and anti--HBs monoclonal antibody coupling, chemical luminous substrate is outside hydrogen peroxide, the NaOH, and all the other all prepare the hepatitis b virus preS 2 antigen chemiluminescence immune analysis determination kit with the method identical with embodiment 1.
The using method of embodiment 7 kits of the present invention
The concrete operations of the hepatitis b virus preS 2 antigen chemiluminescence immune analysis determination kit of above embodiment 1 preparation are as follows:
1) in 4 ℃ of refrigerators, takes out kit, equilibrium at room temperature 15 minutes.
2) take out coated slab, insert on the grillage.
3) blank 1 hole is established in each test, respectively each concentration point calibration object 0,1,5,10,50,100ng/ml and sample to be tested 50 μ L are added corresponding micropore, every hole adds biotinylated antibody 50 μ L except that blank well then, the mixing that fully vibrates, 37 ℃ of incubations 30 minutes.
4) get rid of dereaction liquid, the cleansing solution after the dilution is filled it up with in every hole, washes plate 5 times, buckles on clean thieving paper at last and does.
5) except that blank well, every hole adds 100 μ L respectively anti--HBs label, 37 ℃ of incubations 30 minutes.
6) get rid of dereaction liquid, the cleansing solution after the dilution is filled it up with in every hole, washes plate 5 times, buckles on clean thieving paper at last and does.
7) each hole adds chemical luminous substrate liquid 100 μ L, and fully vibrating with micro-oscillator mixes, room temperature (20～25 ℃) lucifuge reaction 30 minutes.
8) must measure in the 30th～60 minute after adding chemical luminous substrate liquid, on the chemiluminescence measuring instrument, measure the luminous intensity (RLU) in each hole in regular turn, 1 second/hole of Measuring Time.
9) be horizontal ordinate with calibration object concentration, the RLU value is drawn typical curve for ordinate, finds the concentration of the hepatitis b virus preS 2 antigen of this serum on typical curve with each test serum RLU value.
The parallel contrast test of embodiment 8 kits of the present invention and ELISA reagent
Normal control sample 106 examples are made a definite diagnosis hepatitis B great three positive sample 89 examples, hepatitis B "small three positive" sample 67 examples.
Adopt kit of the present invention (chemoluminescence method) to carry out parallel detection with hepatitis B preS 2 antigen detectable (euzymelinked immunosorbent assay (ELISA)), trace routine and result's judgement are carried out in strict accordance with each reagent instructions.
Above-mentioned two kinds of hepatitis B preS 2 antigen detection kit are carried out accuracy, specificity, sensitivity, accuracy performance index evaluations such as (recovery) respectively.
Testing result is shown in table 1-2:
The performance index evaluation of two kinds of hepatitis B preS 2 antigens of table 1 detectable
The testing result of two kinds of hepatitis B preS 2 antigens of table 2 detectable gathers
The described data of table 1-2 show that kit accuracy of the present invention, specificity, the recovery are better than hepatitis B preS 2 antigen enzyme-linked immunologic detecting kit, and sensitivity significantly is better than enzyme-linked immunologic detecting kit.The coincidence rate of two kinds of detection methods of kit of the present invention and enzyme-linked immunologic detecting kit is 98.09%.
Claims (10)
1, a kind of hepatitis b virus preS 2 antigen chemiluminescence immune analysis determination kit, it is characterized in that described kit is made up of the monoclonal antibody of the solid phase carrier of hepatitis b virus preS 2 antigen calibration object, Avidinization, biotinylated hepatitis b virus preS 2 antigen, anti--HBs labeling of monoclonal antibody thing, chemical luminous substrate and concentrated cleaning solution.
2, kit as claimed in claim 1 is characterized in that, described solid phase carrier is microwell plate, plastic tube or magnetic-particle.
3, kit as claimed in claim 1 is characterized in that, described label is alkaline phosphatase or chemiluminescence agent.
4, kit as claimed in claim 3 is characterized in that, described chemiluminescence agent is acridinium ester, luminol or different luminol.
5, kit as claimed in claim 1 is characterized in that, described chemical luminous substrate is 1,2-two oxidative ethane analog derivatives, urea peroxide or hydrogen peroxide.
6, a kind of method for preparing the described kit of claim 1 is characterized in that may further comprise the steps:
1) with the pure product preparation of hepatitis b virus preS 2 antigen calibration object;
2) with the Avidin bag by solid phase carrier;
3) make the monoclonal antibody biotinylation of hepatitis b virus preS 2 antigen;
4) resist-the HBs monoclonal antibody with the label mark;
5) preparation chemical luminous substrate;
6) preparation concentrated cleaning solution;
7) the above-mentioned calibration object of packing, biotinylated antibody, label, chemical luminous substrate and concentrated cleaning solution; And
8) be assembled into finished product.
7, method as claimed in claim 6, it is characterized in that, described with the Avidin bag by the step 2 of solid phase carrier) adopt following method: with 0.050M pH value is the Avidin coating buffer that 9.6 carbonate buffer solution is mixed with desired concn, and coating buffer is carried on the solid phase carrier, with the physiological saline washing, after the sealing of 10% lowlenthal serum, the dehumidifier drying, aluminium foil bag is airtight, cryopreservation.
8, method as claimed in claim 6 is characterized in that, described label is alkaline phosphatase or chemiluminescence agent.
9, method as claimed in claim 8 is characterized in that, described chemiluminescence agent is acridinium ester, luminol or different luminol.
10, method as claimed in claim 6 is characterized in that, described chemical luminous substrate is urea peroxide or hydrogen peroxide.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102565032A (en) * | 2010-12-15 | 2012-07-11 | 北京勤邦生物技术有限公司 | Magnetic granule chemiluminescence kit for detecting nitrofurantoin metabolite and application of magnetic granule chemiluminescence kit |
| CN102565405A (en) * | 2011-08-24 | 2012-07-11 | 苏州长光华医生物试剂有限公司 | Method for immunological detection by combining acridinium ester labeling technology with general magnetic particles |
| CN108271358A (en) * | 2014-12-10 | 2018-07-10 | 财团法人卫生研究院 | Judge the antibody lacked in HBV virus pre-S2 regions and its method |
| CN108344869A (en) * | 2018-02-05 | 2018-07-31 | 苏州长光华医生物医学工程有限公司 | A kind of hepatitis B surface antigen chemiluminescence immune detection reagent kit and its application |
| CN111175516A (en) * | 2020-01-06 | 2020-05-19 | 潍坊市康华生物技术有限公司 | Vascular endothelial growth factor detection kit and preparation method thereof |
| CN117491650A (en) * | 2023-11-01 | 2024-02-02 | 首都医科大学宣武医院 | Quantitative detection kit, detection method and application of peripheral blood Hb-Abeta complex |
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2008
- 2008-03-25 CN CN 200810102665 patent/CN101377503A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102565032A (en) * | 2010-12-15 | 2012-07-11 | 北京勤邦生物技术有限公司 | Magnetic granule chemiluminescence kit for detecting nitrofurantoin metabolite and application of magnetic granule chemiluminescence kit |
| CN102565405A (en) * | 2011-08-24 | 2012-07-11 | 苏州长光华医生物试剂有限公司 | Method for immunological detection by combining acridinium ester labeling technology with general magnetic particles |
| CN108271358A (en) * | 2014-12-10 | 2018-07-10 | 财团法人卫生研究院 | Judge the antibody lacked in HBV virus pre-S2 regions and its method |
| CN108344869A (en) * | 2018-02-05 | 2018-07-31 | 苏州长光华医生物医学工程有限公司 | A kind of hepatitis B surface antigen chemiluminescence immune detection reagent kit and its application |
| CN111175516A (en) * | 2020-01-06 | 2020-05-19 | 潍坊市康华生物技术有限公司 | Vascular endothelial growth factor detection kit and preparation method thereof |
| CN117491650A (en) * | 2023-11-01 | 2024-02-02 | 首都医科大学宣武医院 | Quantitative detection kit, detection method and application of peripheral blood Hb-Abeta complex |
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Owner name: BEIJING KEMEI BIOLOGICAL TECHNOLOGY CO., LTD. Free format text: FORMER OWNER: KEMEI DONGYA BIOLOGICAL TECHNOLOGY CO., LTD., BEIJING Effective date: 20111028 |
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Effective date of registration: 20111028 Address after: 100094 Beijing city Haidian District Yongfeng base Feng Xian Road No. 7 North Park Applicant after: Beijing Kemei Biological Technology Co., Ltd. Address before: 100094 Beijing city Haidian District Yongfeng base Feng Xian Road No. 7 North Park Applicant before: Kemei Dongya Biological Technology Co., Ltd., Beijing |
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Application publication date: 20090304 |