CN109444414A - Using the method and corresponding composition, kit of AlphaLISA detection dengue virus - Google Patents
Using the method and corresponding composition, kit of AlphaLISA detection dengue virus Download PDFInfo
- Publication number
- CN109444414A CN109444414A CN201811048274.2A CN201811048274A CN109444414A CN 109444414 A CN109444414 A CN 109444414A CN 201811048274 A CN201811048274 A CN 201811048274A CN 109444414 A CN109444414 A CN 109444414A
- Authority
- CN
- China
- Prior art keywords
- antibody
- detection
- alphalisa
- dengue virus
- control product
- 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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
-
- 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
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Hematology (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Pathology (AREA)
- Virology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Cell Biology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Tropical Medicine & Parasitology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention provides the methods and corresponding composition, kit of application AlphaLISA detection dengue virus.The AlphaLISA detection combination object, comprising: be coated with the Acceptor beads of capture antibody；The detection antibody of biotin labeling；It is coated with the donor microballon of Streptavidin；The capture antibody and detection antibody are the pairing antibody of a pair of of anti-dengue virus NS1 albumen.It, as antigen, has good specificity and sensitivity for NS1 albumen.A kind of AlphaLISA kit for detecting dengue virus is also disclosed simultaneously comprising above-mentioned AlphaLISA detection combination object and 384 hole microwell plates, positive quality control product and negative quality-control product；The positive quality control product is the matrix serum of the albumen containing dengue virus NS 1, and the feminine gender quality-control product is the solution containing basis buffer.The kit is able to use the more microwell plate of hole count and is detected, and realizes high-throughput large sample fast qualitative detection dengue virus.
Description
Technical field
The invention belongs to field of biomedicine technology, are related to medicine, immunology, microbiology, pestology, and in particular to
A kind of dengue virus NS 1 albumin A lphaLISA detection method.
Background technique
Dengue fever (dengue fever) be dengue virus through mosquito matchmaker propagate caused by acute human insect-borne infectious disease,
It is widely current in world wide.It can lead to subclinical infection, dengue fever, dengue hemorrhagic fever after dengue virus infection human body.Typically
Dengue fever clinical manifestation is that onset is hurried, and high fever, headache, muscle, Bones and joints are acutely ached, fash occur in some patientss, bleeding
Tendency, enlargement of lymph nodes, white blood cell count(WBC) reduction, decrease of platelet etc..This disease is mainly popular in subtropical and tropical zones, I
The ground such as state Guangdong, Hong Kong, Macao are the Endemic Areas of dengue fever.Since this disease is to be propagated by yellow-fever mosquito, therefore prevalence has certain season
Property, generally in 5 annual~November, peak is in 7~September part.In new Endemic Area, crowd is generally susceptible, but falls ill with adult
Based on, in endemic area, fall ill based on children.Due to lacking the measure of effective control mosquito, dengue fever is in prevalence
Still there is higher disease incidence in season, such as breaks out large area Dengue epidemic situation in Guangzhou within 2014, there is hundreds of new disease daily
Example occurs, and there are also deaths, cause tremendous influence to people's health.Therefore, exploitation is directed to the dengue virus of large sample
Rapid detection method, for epidemic situation control and clinical treatment and prognosis important role.
Currently, the laboratory diagnosis of dengue fever mainly has isolation of virus, nucleic acid amplification and serology antibody or antigen
Detection method.Virus purification result is accurate and reliable but time-consuming too long, and sensitivity is low, it is impossible to be used in large sample screening.Nucleic acid amplification is logical
Often there are RT-PCR and fluorescence RT-PCR method, but be confined to the viremia virusemia phase, it is also desirable to extract nucleic acid and take a lot of work than relatively time-consuming.Serology
There are detection specific IgM antibodies, but detection window will be after the 5d that falls ill, and exists with the infection of other flaviviridaes and intersect instead
It answers.Therefore, antigen detection is developing direction.
NS1 albumen is one of the non-structural protein of 7 kinds of dengue virus, highly conserved in 4 kinds of serotypes of virus, is energy
It reaches in infection cell surface expression and is secreted into extracellular non-structural protein.Existing studies have shown that is in morbidity early stage, human serum
In in the case where can't detect IgM antibody or RNA, but can detecte viral antigen NS1 albumen.As it can be seen that NS1 albumen is being stepped on
There is important value in the especially dengue fever early diagnosis of leather viral diagnosis.For NS1 antigen, there are many researchers to establish at present
The ELISA detection method of double antibodies sandwich, and obtain preferable specificity and sensitivity.But ELISA method operating procedure is numerous
It is trivial, with duration, multistep is needed to wash, it is difficult to adapt to large sample detection.
Alpha (Amplified luminescent proximity homogenous assay) technology is one and passes through
Singlet oxygen molecular transmitting carrys out the homogeneous immunologic detection method of enhanced chemiluminescence.Popular says exactly there is 2 in reaction solution
Kind of microballon, it is a kind of can be photosensitive, it is a kind of to shine, when immunology antigen-antibody reaction makes this 2 kinds of microballons at reference state and mutually
Within close to certain distance after (200nm), if the photosensitive microballon of excitation, photosensitive microballon can be by discharging singlet
Oxonium ion transfers energy to luminous microballon, and by certain chemical reactions, the microballon that shines then can emit more shortwave with more high level
Long light.People are used using AlphaScreen the and AlphaLISA technology that Alpha technology establishes similar ELISA at present
In detection protein, cell factor even small molecule etc..Compared with traditional ELISA (ELISA) method, it has
The advantages that homogeneous reaction is not necessarily to wash, easy to operate, the following alternative part ELISA detection reagent, application prospect very may be used
It sees.
Summary of the invention
The present invention is directed to the high-throughput large sample detection method for establishing dengue virus using AlphaLISA technology platform is concurrent
Exhibition is kit, can provide novel quick detection technique with the prevention and control of public health arboviruse.It specially discloses a kind of dual anti-
The sandwich AlphaLISA of body detects dengue virus method.
To achieve the goals above, using following technical scheme:
A kind of AlphaLISA detection combination object, comprising:
(1) Acceptor beads of capture antibody are coated with；
(2) the detection antibody of biotin labeling；
(3) it is coated with the donor microballon of Streptavidin；
Wherein, the capture antibody and detection antibody are the pairing antibody of a pair of of anti-dengue virus NS1 albumen.
The Acceptor beads that (1) is coated with capture antibody are obtained by following making step:
1-1. carries out the purification process that desalts to capture antibody with desalination centrifugal column, and adjusting antibody concentration is 1mg/mL；
1-2. is micro- to the AlphaLISA receptor that 100 μ L step 1-1 antibody addition obtained, 50 μ L concentration are 20mg/mL
10% Tween-20, the NaBH of the 400mM of 10 μ L of 1.25 μ L is then added in pearl suspension3CN solution, with the pH of 100mM
7.4 Hepes buffer is supplied to 200 μ L final volumes, and 37 DEG C of rotations, which are swayed, to be incubated for 20 hours；
The CMO solution that the 10 freshly prepared concentration of μ L are 65mg/mL is added in 1-3. after the completion of being incubated for, 37 DEG C are incubated for 1 hour；
1-4.PBS washing centrifugation 2~3 times, Acceptor beads are resuspended in ultrasonic vibration.
Further, it is described be coated with capture antibody Acceptor beads using final concentration of 10 μ g/mL.
The detection antibody of (2) biotin labeling is obtained by following making step:
2-1. purification assays antibody, adjusting concentration is 1mg/mL, pH >=7.0, and the NHS-ChromaLink- of 2mg/mL is added
Biotin biotin labeling reagent solution, 23 DEG C are incubated for 2 hours；
2-2. is centrifuged the mixed solution of column purification step 2-1 with desalination.
Further, the detection antibody of the biotin labeling uses final concentration of 5 μ g/mL.
(3) be coated with the donor microballon of Streptavidin using final concentration of 40 μ g/mL.
The present invention also discloses a kind of AlphaLISA kits for detecting dengue virus comprising above-mentioned
AlphaLISA detection combination object and 384 hole microwell plates, positive quality control product and negative quality-control product；The positive quality control product be containing
The matrix serum of dengue virus NS 1 albumen, the feminine gender quality-control product is the solution containing basis buffer.
The application method of the AlphaLISA kit, comprising steps of sample to be examined or the positive is added to 384 hole microwell plates
Quality-control product or negative quality-control product, are then added the Acceptor beads for being coated with capture antibody, then add biotin labeling detection it is anti-
Body, is incubated for 30 at 23 DEG C~be added be coated with the donor microballon of Streptavidin after sixty minutes, it is incubated for 20 again in dark situation~
30 minutes, then detect fluorescent value.
Chinese invention patent application (application No. is: 201210031802.X, 201210047016.9) it individually discloses and adopts
With the side of AlphaLISA technology detection enterovirns type 71 capsid protein (Anti-EV71VP1IgM) and hepatitis A disease IgM
Method.But it there is no the technical solution of the kit and detection method using AlphaLISA technology detection dengue virus at present.
AlphaLISA detection combination object of the present invention, as antigen, has good specificity and spirit for NS1 albumen
Sensitivity.
It is carried out by the microwell plate that kit prepared by AlphaLISA detection combination object is able to use hole count more (384 hole)
Detection, far more than common 96 hole microwell plate, may be implemented high-throughput large sample fast qualitative detection dengue virus.
Detailed description of the invention
Fig. 1 is the Linear correlative analysis result figure for detecting the AlphaLISA kit of dengue virus.
Specific embodiment
Below with reference to embodiment, the present invention is further illustrated, and however, it is not limited to this.It is adopted in following embodiment
Molecular biology experiment technology includes PCR amplification, plasmid extracts, plasmid converts, DNA fragmentation connection, digestion, gel electrophoresis
Deng unless otherwise specified, usually conventionally operating, for details, reference can be made to " Molecular Cloning:A Laboratory guide " (third editions)
(Sambrook J, Russell DW, Janssen K, Argentine J. Huang Peitang etc. is translated, and 2002, Beijing: scientific publication
Society), or according to the normal condition proposed by manufacturer.
Embodiment 1: the preparation of the AlphaLISA kit of dengue virus is detected.
By the capture antibody for matching antibody each other and detects the buffer exchange of antibody and purifying and remove Sodium azide to desalt
(NaN3).By the Zeba of Thermo Fisher Scientific companyTMCentrifugation goes salt plug to be put into corresponding collecting pipe, 1500g from
Heart 1min removal storage liquid.It is washed 2 times with 1 × PBS again.Antibody stoste is added and puts on new collecting pipe, 1500g centrifugation
1min retains the antibody-solutions in collecting pipe, is prepared into the antibody concentration of 1mg/ml.
Detect antibody coupling biotin.By purified 1mg/mL, the NHS- of 2mg/mL is added in the detection antibody of pH >=7.0
Volume is simultaneously adjusted to 200uL by ChromaLink-biotin biotin labeling reagent solution, and 23 DEG C are incubated for 2 hours, then use
ZebaTMDesalination centrifugal column collects the antibody of marked biotin after purification, and concentration is about 500ug/mL.Recommend eventually when practical application
Concentration is 5ug/mL.
Acceptor beads coating capture antibody.0.1mg capture antibody is added to the Acceptor beads of AlphaLISA containing 1mg (50 μ
L, 20mg/mL), the 10%Tween-20 of 1.25 μ L is then added, the NaBH3CN solution of the 400mM of 10 μ L finally uses Hepes
Buffer (100mM, pH 7.4) is supplied to 200 μ L final volumes, and 37 DEG C of rotations, which are swayed, to be incubated for 20 hours.10 are added after the completion of being incubated for
The freshly prepared confining liquid of μ L (the CMO solution of 65mg/mL), 37 DEG C are incubated for 1 hour to close unreacted site.Last PBS
Washing centrifugation 2-3 times, 4 DEG C of refrigerations are protected from light spare, concentration 5mg/mL after microballon is resuspended with ultrasound.Recommend when practical application dense eventually
Degree is 10ug/mL.
It prepares immune detection buffer: being that mother liquor is separately added into other ingredients with 25mmol/L HEPES (pH7.4), at
Point have 50mmol/L NaCI, 10mmol/L DTPA, 0.5%BSA, 2mg/mL Dextran-500,0.1%Tween-20,
0.02% bovineγ-globulin, 0.01%Proclin-300,0.01% gentamicin, various reagents need to sufficiently dissolve, finally mistake
Bacterium is filtered out, is saved in 4 DEG C.
It prepares standard items buffer: being that mother liquor is separately added into other ingredients with 50mmol/L Tris-HCl (pH7.8),
Ingredient has 0.9%NaCI, 1.5%BSA, 0.01%Tween-20,0.05%Proclin-300, and the buffer prepared needs to filter
Degerming is saved in 4 DEG C.
Prepare positive quality control product: after dissolution dengue virus NS 1 albumen, gradient dilution is respectively: 400ng/ at 6 concentration
ml、200ng/ml、100ng/ml、50ng/ml、20ng/ml、10ng/ml。
Embodiment 2: the evaluation test of the AlphaLISA kit of dengue virus is detected.
Detection reagent: related reagent preparation is carried out according to 1 the method for embodiment.
Detecting step: sample to be examined or positive quality control product or negative quality-control product is added to 384 hole microwell plates, packet is then added
The Acceptor beads for being had capture antibody, then add the detection antibody of biotin labeling, it is incubated for 30 at 23 DEG C~it is added after sixty minutes
It is coated with the donor microballon of Streptavidin, is incubated for again in dark situation 20~30 minutes, then detects fluorescent value.
1, Linear correlative analysis
The positive quality control product of gradient dilution and negative quality-control product are detected simultaneously with the method for the invention, with sample
It is abscissa that concentration, which takes logarithm, and fluorescence signal value is ordinate, and linear curve obtains dengue virus NS 1 Protein Detection side
Response curve and linearly dependent coefficient r value (as shown in Figure 1) between method and sample dose.The r of test display for several times2> 0.98, table
Bright good relationship.
2, the determination of CUT-OFF value
The present invention is qualitative checking method.Take the normal population serum sample and 10 parts of feminine genders of 350 parts of dengue virus feminine genders
Quality-control product is detected with the method for the invention.Count the ratio of normal person's pattern detection fluorescent value and negative quality-control product mean value
Value, the average value of ratio calculated are 1.52, and standard deviation (SD) is 0.27.Add two SD with ratio mean value, primarily determines as we
The CUT-OFF value of method is 2.06, determines method then referring to the more general CUF-OFF value of numerous clinical detection reagent producers, most
Eventually it is proposed that CUT-OFF value are as follows: CUT-OFF=negative control fluorescent value × 2.1.As detection sample fluorescence Zhi≤CUT-
OFF is then judged to the positive, is otherwise feminine gender.
3, specificity analysis
Negative samples of 55 parts of the selection without dengue virus are detected, including normal human serum, influenza virus infection
Patients serum, flavirirus vaccines strain, zika virus culture, chikungunya virus culture etc..The results show that 53 parts of sample inspections
It surveys to be negative, 2 parts are determined as the positive, and calculating specificity is 96.4%.
4, Precision Analyze
Positive quality control product is prepared by high, medium and low 3 different level concentration, each parallel 3 hole respectively carries out 10 repetitions
Property test.Its coefficient of variation CV is calculated, as a result within the scope of 4.37-7.45%, shows that precision is preferable.
Embodiment 3: benchmark test
Method one is the AlphaLISA method that the present invention establishes.Method two is commercial kit (cortez company, the U.S.
Dengue fever NS1 quick detection reagent), detection sensitivity 6ng/mL.
Sample: 100 parts of clinical serum samples of random selection have Dengue positive case, also have normal person, while using above-mentioned
2 kinds of methods are detected.
Its yin and yang attribute coincidence rate of post analysis and consistent degree percentage are completed in detection, as a result as described in Table 1.
1 qualitative checking method Evaluation results of table
It is 95.38% according to the positive coincidence rate that test result calculates, negative match-rate 94.29%, consistent degree hundred
Divide than being 95%.
Conclusion: the AlphaLISA kit and its application method of detection dengue virus provided by the present invention and commercialization
Kit coincidence rate it is higher, be able to satisfy requirement that is high-throughput, quickly detecting, there is good commercial value.
Claims (9)
- Application of the 1.AlphaLISA detection method in detection dengue virus.
- 2. a kind of AlphaLISA detection combination object, characterized by comprising: be coated with Acceptor beads, the biotin of capture antibody The detection antibody of label, the donor microballon for being coated with Streptavidin；The capture antibody and detection antibody are a confrontation Dengues The pairing antibody of virus N S1 albumen.
- 3. AlphaLISA detection combination object according to claim 2, which is characterized in that described be coated with captures antibody Acceptor beads are obtained by following making step:1-1. carries out the purification process that desalts to capture antibody with desalination centrifugal column, and adjusting antibody concentration is 1mg/mL；1-2. is added the AlphaLISA Acceptor beads that 50 μ L concentration are 20mg/mL to 100 μ L step 1-1 antibody obtained and hangs 10% Tween-20, the NaBH of the 400mM of 10 μ L of 1.25 μ L is then added in supernatant liquid3CN solution, with the pH 7.4 of 100mM Hepes buffer supply to 200 μ L final volumes, incubation 20 hours is swayed in 37 DEG C of rotations；The CMO solution that the 10 freshly prepared concentration of μ L are 65mg/mL is added in 1-3. after the completion of being incubated for, 37 DEG C are incubated for 1 hour；1-4.PBS washing centrifugation 2~3 times, Acceptor beads are resuspended in ultrasonic vibration.
- 4. AlphaLISA detection combination object according to claim 3, which is characterized in that described be coated with captures antibody Acceptor beads use final concentration of 10 μ g/mL.
- 5. AlphaLISA detection combination object according to claim 2, which is characterized in that the detection of the biotin labeling Antibody is obtained by following making step:2-1. purification assays antibody, adjusting concentration is 1mg/mL, pH >=7.0, and the NHS-ChromaLink- of 2mg/mL is added Biotin biotin labeling reagent solution, 23 DEG C are incubated for 2 hours；2-2. is centrifuged the mixed solution of column purification step 2-1 with desalination.
- 6. AlphaLISA detection combination object according to claim 5, which is characterized in that the detection of the biotin labeling Antibody uses final concentration of 5 μ g/mL.
- 7. AlphaLISA detection combination object according to claim 2, which is characterized in that described to be coated with Streptavidin Donor microballon use final concentration of 40 μ g/mL.
- 8. a kind of AlphaLISA kit for detecting dengue virus, it is characterised in that including any one of such as claim 2~7 institute AlphaLISA detection combination object and 384 hole microwell plates, positive quality control product and the negative quality-control product stated；The positive quality control product For the matrix serum of the albumen containing dengue virus NS 1, the feminine gender quality-control product is the solution containing basis buffer.
- 9. detecting the application method of the AlphaLISA kit of dengue virus described in claim 8, it is characterised in that including step It is rapid: sample to be examined or positive quality control product or negative quality-control product is added to 384 hole microwell plates, is then added and is coated with capture antibody Acceptor beads, then add the detection antibody of biotin labeling are incubated for 30 at 23 DEG C~are added that be coated with strepto- affine after sixty minutes The donor microballon of element, is incubated for 20~30 minutes again in dark situation, then detects fluorescent value.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811048274.2A CN109444414A (en) | 2018-09-10 | 2018-09-10 | Using the method and corresponding composition, kit of AlphaLISA detection dengue virus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811048274.2A CN109444414A (en) | 2018-09-10 | 2018-09-10 | Using the method and corresponding composition, kit of AlphaLISA detection dengue virus |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109444414A true CN109444414A (en) | 2019-03-08 |
Family
ID=65530258
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811048274.2A Pending CN109444414A (en) | 2018-09-10 | 2018-09-10 | Using the method and corresponding composition, kit of AlphaLISA detection dengue virus |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109444414A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112255399A (en) * | 2020-10-20 | 2021-01-22 | 浙江洪晟生物科技股份有限公司 | Hog cholera virus detection kit based on double-antibody sandwich homogeneous phase chemiluminescence method, and preparation method and application thereof |
| CN112326635A (en) * | 2020-10-20 | 2021-02-05 | 浙江理工大学 | Double-antibody sandwich homogeneous-phase chemiluminescence porcine circovirus type 2 detection kit, and preparation method and application thereof |
| CN113092782A (en) * | 2021-03-31 | 2021-07-09 | 北京大学 | Method for screening chemical nuclear receptor activity based on Alpha technology high-throughput and multi-target |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101915836A (en) * | 2010-07-23 | 2010-12-15 | 中国检验检疫科学研究院 | Protein suspension chip for detecting dengue antibody in serum sample and preparation method and using method thereof |
| CN102161983A (en) * | 2011-03-28 | 2011-08-24 | 中山大学 | Hybridoma cell line secreting I-IV-type dengue virus NS1 monoclonal antibodies and test kit thereof |
| CN102590505A (en) * | 2012-02-13 | 2012-07-18 | 中国疾病预防控制中心病毒病预防控制所 | Reagent kit for detecting immunoglobulin m (IgM) AlphaLISA for resisting enterovirus71 capsid protein1 |
| CN102608313A (en) * | 2012-02-27 | 2012-07-25 | 中国疾病预防控制中心病毒病预防控制所 | Anti-hepatitis A virus IgM (immunoglobulin M) AlphaLISA detection kit |
| CN103235131A (en) * | 2013-03-22 | 2013-08-07 | 深圳国际旅行卫生保健中心 | Lateral flow immunochromatographic determination product for detecting yellow fever viruses and preparation method of lateral flow immunochromatographic determination product |
| CN103529181A (en) * | 2013-10-11 | 2014-01-22 | 重庆出入境检验检疫局检验检疫技术中心 | AlphaLISA detection kit of penicillinase in milk product |
| WO2015196192A2 (en) * | 2014-06-20 | 2015-12-23 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Methods and compositions relating to dengue virus |
| CN105759046A (en) * | 2016-04-08 | 2016-07-13 | 重庆理工大学 | Newcastle disease virus double-antibody sandwich AlphaLISA detection kit and detection method thereof |
| CN106290847A (en) * | 2015-05-13 | 2017-01-04 | 上海凯创生物技术有限公司 | A kind of dengue virus NS1 antigen emulsion technique detection kit |
| US20170067895A1 (en) * | 2011-12-22 | 2017-03-09 | Inbios International, Inc. | Methods and materials for the detection of dengue virus infection |
| CN107543923A (en) * | 2017-08-23 | 2018-01-05 | 黑龙江省动物疫病预防与控制中心 | Detect the kit and its detection method of avian leukosis virus A/B/J subgroup specific antibodies |
| WO2018106945A1 (en) * | 2016-12-07 | 2018-06-14 | Progenity Inc. | Gastrointestinal tract detection methods, devices and systems |
-
2018
- 2018-09-10 CN CN201811048274.2A patent/CN109444414A/en active Pending
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101915836A (en) * | 2010-07-23 | 2010-12-15 | 中国检验检疫科学研究院 | Protein suspension chip for detecting dengue antibody in serum sample and preparation method and using method thereof |
| CN102161983A (en) * | 2011-03-28 | 2011-08-24 | 中山大学 | Hybridoma cell line secreting I-IV-type dengue virus NS1 monoclonal antibodies and test kit thereof |
| US20170067895A1 (en) * | 2011-12-22 | 2017-03-09 | Inbios International, Inc. | Methods and materials for the detection of dengue virus infection |
| CN102590505A (en) * | 2012-02-13 | 2012-07-18 | 中国疾病预防控制中心病毒病预防控制所 | Reagent kit for detecting immunoglobulin m (IgM) AlphaLISA for resisting enterovirus71 capsid protein1 |
| CN102608313A (en) * | 2012-02-27 | 2012-07-25 | 中国疾病预防控制中心病毒病预防控制所 | Anti-hepatitis A virus IgM (immunoglobulin M) AlphaLISA detection kit |
| CN103235131A (en) * | 2013-03-22 | 2013-08-07 | 深圳国际旅行卫生保健中心 | Lateral flow immunochromatographic determination product for detecting yellow fever viruses and preparation method of lateral flow immunochromatographic determination product |
| CN103529181A (en) * | 2013-10-11 | 2014-01-22 | 重庆出入境检验检疫局检验检疫技术中心 | AlphaLISA detection kit of penicillinase in milk product |
| WO2015196192A2 (en) * | 2014-06-20 | 2015-12-23 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Methods and compositions relating to dengue virus |
| CN106290847A (en) * | 2015-05-13 | 2017-01-04 | 上海凯创生物技术有限公司 | A kind of dengue virus NS1 antigen emulsion technique detection kit |
| CN105759046A (en) * | 2016-04-08 | 2016-07-13 | 重庆理工大学 | Newcastle disease virus double-antibody sandwich AlphaLISA detection kit and detection method thereof |
| WO2018106945A1 (en) * | 2016-12-07 | 2018-06-14 | Progenity Inc. | Gastrointestinal tract detection methods, devices and systems |
| CN107543923A (en) * | 2017-08-23 | 2018-01-05 | 黑龙江省动物疫病预防与控制中心 | Detect the kit and its detection method of avian leukosis virus A/B/J subgroup specific antibodies |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112255399A (en) * | 2020-10-20 | 2021-01-22 | 浙江洪晟生物科技股份有限公司 | Hog cholera virus detection kit based on double-antibody sandwich homogeneous phase chemiluminescence method, and preparation method and application thereof |
| CN112326635A (en) * | 2020-10-20 | 2021-02-05 | 浙江理工大学 | Double-antibody sandwich homogeneous-phase chemiluminescence porcine circovirus type 2 detection kit, and preparation method and application thereof |
| CN113092782A (en) * | 2021-03-31 | 2021-07-09 | 北京大学 | Method for screening chemical nuclear receptor activity based on Alpha technology high-throughput and multi-target |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111693712B (en) | Method for detecting SARS-CoV-2N protein of new coronavirus by using nucleic acid aptamer | |
| Liu et al. | Highly specific detection of thrombin using an aptamer-based suspension array and the interaction analysis via microscale thermophoresis | |
| US8785132B2 (en) | Aptamer sandwich assays | |
| CN109444414A (en) | Using the method and corresponding composition, kit of AlphaLISA detection dengue virus | |
| US20230228751A1 (en) | Aptamers against sars-cov-2 | |
| WO2017008179A1 (en) | Reagent for high throughput combined detection of hepatitis c virus antigen and antibody | |
| CN113195722B (en) | DNA aptamer specifically bound to chikungunya virus E2 and application thereof | |
| CN103033619A (en) | Protein chip reagent kit and method for comprehensively detecting lung cancer marker | |
| CN102965378B (en) | Aptamer of glycosylated hemoglobin and preparation method thereof | |
| CN104634961A (en) | Preparation method and detection method of rabies virus homogeneous fluorescent composite microsphere | |
| CN104364649A (en) | Protein expression analyses for identifying genotoxic compounds | |
| Martínez-Roque et al. | DNA aptamer selection for SARS-CoV-2 spike glycoprotein detection | |
| Ma et al. | A multiple-target simultaneous detection method for immunosorbent assay and immunospot assay | |
| CN113321715A (en) | Novel coronavirus antigen and detection use thereof | |
| CN104515855A (en) | Galectin-3 detection nanometer magnetic bead sorting-time resolved immunofluorescence kit | |
| KR102048812B1 (en) | DNA aptamer binding specifically to dengue virus envelope protein domain III(DENV EDIII) and uses thereof | |
| Xu et al. | Multiplex biomarker analysis biosensor for detection of hepatitis B virus | |
| CN102183666B (en) | Liquid phase chip for detecting twelve pathogen antibodies in blood serum sample in high flux, and preparation method and using method thereof | |
| CN104109674A (en) | Aptamer of glycosylated hemoglobin and preparation method thereof | |
| CN104109675A (en) | Aptamer of glycosylated hemoglobin and preparation method thereof | |
| CN104164436A (en) | Aptamer of glycosylated hemoglobin, and preparation method thereof | |
| CN114410641B (en) | Aptamer for detecting rubella virus, kit and application | |
| CN105483132B (en) | The aptamer of affine viral hepatitis type C core antigen and its application | |
| CN114410640B (en) | Aptamer for detecting measles virus, kit and application | |
| CN114381460B (en) | Aptamer for detecting mumps virus, kit and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for 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: 20190308 |