CN104714028A - Immunodetection method for detecting IgM antibody - Google Patents
Immunodetection method for detecting IgM antibody Download PDFInfo
- Publication number
- CN104714028A CN104714028A CN201510092986.4A CN201510092986A CN104714028A CN 104714028 A CN104714028 A CN 104714028A CN 201510092986 A CN201510092986 A CN 201510092986A CN 104714028 A CN104714028 A CN 104714028A
- Authority
- CN
- China
- Prior art keywords
- igm
- monoclonal antibody
- antibody
- antigen
- solid phase
- 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.)
- Granted
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
- G01N33/68—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
Abstract
The invention discloses an immunodetection method for detecting an IgM antibody, and relates to an immunodetection technology of the IgM antibody. The immunodetection method comprises the following steps: 1, curing processing of anti-IgM; 2, combination of IgM to be detected with the anti-IgM; 3, combination of an antigen and a conjugated monoclonal antibody with the IgM to be detected on a solid phase carrier; and 4, signal detection. The immunodetection method has the advantages of greatly reducing the interference from the IgM to the detection result, reducing the background and false positive rate of the detection, improving the detection specificity, enhancing the detection signal, reducing the false negative rate and improving the detection sensitivity.
Description
Technical field
The present invention relates to technical field of biomedical detection, particularly relate to IgM antibody technical field of immunoassay.
Background technology
In existing immunologic detection method, mainly applied immunology detection technique detects test substance in sample; In clinical examination, detect the material such as antibody or antigen in body fluid mainly through antigen-antibody reaction.According to the difference of Cleaning Principle, immune detection can be divided into indirect method, sandwich method, competition law and prize law.
In normal serum; IgG is the principal ingredient of serum immune globulin; account for 75% of Immunoglobulin in Serum total content; IgM then accounts for greatly 6% of Immunoglobulin in Serum total amount; for certain antigen specific IgM often and specific IgG exist simultaneously, and specific IgG often can disturb the mensuration of IgM antibody.In existing four kinds of detection methods, specific IgG is mainly manifested in as follows to the interference that IgM antibody measures:
When indirect method surveys IgM antibody, by envelope antigen, detect by the pattern that enzyme or other indicants mark two resist, these class methods easily occur that specificity is strong, are subject to the interference of IgG in sample to be tested;
When sandwich method surveys IgM antibody, occur that specificity is strong equally, easily by the interference of IgG, detection sensitivity comparatively prize law is not high, and can vie each other with the antigen of tense marker and the antigen of bag quilt is combined with IgM to be measured, causes detection signal by force or do not have;
When competition law surveys IgM antibody, because IgM antibody molecular weight is comparatively large, not be applicable to very much using the method, and the same with indirect method, its specificity is not strong, and be subject to the interference of IgG in sample to be tested, the IgM antibody used with tense marker not easily obtains.
During current mensuration IgM antibody, main prize law, catching rule is by wrapping by anti-IgM, detecting, namely by the pattern of enzyme or other indicant labelled antigens, first Immunoglobulin IgMs all in serum (comprising specific IgM and non-specific IgM) are all fixed in solid phase, after removing IgG interference, then detect specific IgM to be measured, but this in testing process, easily occur that detection sensitivity is not high, easily cause false negative phenomenon.
Summary of the invention
The specificity existed in immunologic detection method for existing detection IgM antibody is not strong, easily to be subject in sample to be tested IgG interference and detection sensitivity not high, easily there is false-negative problem, the object of the invention is to propose a kind of immunologic detection method detecting IgM antibody to solve foregoing problems.
For reaching above-mentioned technical purpose, present invention employs following technical scheme:
The present invention is a kind of immunologic detection method detecting IgM antibody: comprise following steps:
The solidification process of step one, anti-IgM, is placed on solid phase carrier by anti-IgM, under the condition of 2 DEG C ~ 8 DEG C, hatch 12 hours and more than, then wash 3 ~ 5 times;
Step 2, IgM to be measured are combined with anti-IgM, are added in the solid phase carrier having had cured anti-IgM by the IgM to be measured as sample, hatch 0.5 ~ 1.0 hour, wash 3 ~ 5 times;
Step 3, antigen, the mark monoclonal antibody IgM to be measured on solid phase carrier is combined, can after first by antigen, monoclonal antibody mix with mark, then adds in the solid phase carrier that washs in step 2, in constant temperature oven, 37 DEG C of constant temperatures hatch 0.5 ~ 1 hour, hatch end, wash 3 ~ 5 times; Or antigen, mark monoclonal antibody are added in the solid phase carrier washed in step 2 successively, namely first added by antigen, and then add mark monoclonal antibody, in constant temperature oven, 37 DEG C of condition constant-temperature incubations 0.5 ~ 1 hour, hatch end, wash 3 ~ 5 times, wait until detection;
Step 4, input, after step 3 completes, according to the difference of the indicant on mark monoclonal antibody, corresponding instrument and equipment carries out its reaction signal detection.
More preferably, described antigen, mark monoclonal antibody and IgM to be measured hatch in process, antigen share is sub can simultaneously in conjunction with the mark monoclonal antibody of 2 and above different binding site, to increase reaction signal.
More preferably, described solid phase carrier is microwell plate, glass plate or NC film.
More preferably, described washing process adopts the cleansing solution containing surfactant.
More preferably, described anti-IgM is goat-anti IgM or mouse-anti IgM.
More preferably, the indicant in described mark monoclonal antibody is enzyme, collaurum, collargol, electroselenium, latex, radiomaterial, chemiluminescent substance and fluorescent material.
Beneficial effect of the present invention: first the present invention eliminates the combination of IgG and IgM to be measured competitiveness and antigen in sample, greatly reduce the interference of IgG to testing result, thus reduce the background and false positive rate that detect, improve detection specificity, adopt the mark monoclonal antibody of 2 and above different binding site simultaneously, strengthen detection signal, reduce false negative rate, improve detection sensitivity.
Accompanying drawing explanation
Fig. 1 is a kind of reaction principle schematic diagram detecting the immunologic detection method of IgM antibody of the present invention.
Wherein, 1-solid phase carrier; The anti-IgM of 2-; IgM to be measured in 3-sample; 4-antigen; 5-marks monoclonal antibody.
Embodiment
In order to better the present invention is described, be now described further with accompanying drawing in conjunction with the embodiments.
The present invention is a kind of immunologic detection method detecting IgM antibody, reaction principle as shown in Figure 1, and the step related to is:
The solidification process of step one, anti-IgM2.Anti-IgM2 is placed on the microwell plate of detection system or glass plate or NC film, these microwell plates or glass plate or NC film are the solid phase carrier 1 of experiment, under the condition of 2 DEG C ~ 8 DEG C, hatch 12 hours and more than, then wash 3 ~ 5 times with the cleansing solution containing surfactant, the object of washing mainly removes the anti-IgM2 not yet combining upper solid phase carrier 1.
Step 2, IgM3 to be measured are combined with anti-IgM2.Have cured in the solid phase carrier 1 of anti-IgM2 by adding containing IgM3 sample to be measured, hatch 0.5 ~ 1.0 hour, after the anti-IgM2 on IgM3 to be measured and solid phase carrier 1 combines, wash 3 ~ 5 times with the cleansing solution containing surfactant, to remove the specific IgG in sample, avoid specific IgG to the interference detected.
Step 3, antigen, the mark monoclonal antibody IgM3 to be measured on solid phase carrier is combined.Can antigen be marked after monoclonal antibody mixes with at least 2, add in the solid phase carrier that attached to anti-IgM2 and IgM3 to be measured, in constant temperature oven, thermostat temperature 37 DEG C, hatch 0.5 ~ 1 hour, hatch end, adopt the cleansing solution containing surfactant to wash 3 ~ 5 times; Or can first antigen be added in the constant temperature oven being placed on 37 DEG C attached to anti-IgM2 tie and sample zhogn IgM3 to be measured solid phase carrier in, and then at least 2 mark monoclonal antibodies are added, hatch 0.5 ~ 1 hour equally, hatch end, the cleansing solution containing surfactant is adopted to wash 3 ~ 5 times, obtain hatching thing, to be detected;
Step 4, input.After step 3 completes, according to the difference of indicant on mark monoclonal antibody, corresponding instrument and equipment carries out its reaction signal detection.
Particularly, the mark monoclonal antibody that the present invention relates to is 2 and has the monoclonal antibody of different binding site above.
Particularly, anti-IgM used in the present invention can be goat-anti IgM or mouse-anti IgM.
Particularly, the present invention can be applied to euzymelinked immunosorbent assay (ELISA), fast detection method, radio-immunoassay, immunofluorescence assay.
Particularly, indicant of the present invention can be enzyme, collaurum, collargol, electroselenium, latex, radiomaterial, chemiluminescent substance and fluorescent material.
Specific embodiment:
Rubella virus IgM antibody detection kit (euzymelinked immunosorbent assay (ELISA)) is adopted in the present embodiment.
The preparation of 1.1 microwell plates
Bag quilt: by anti-IgM (be specially goat-anti people IgM, be purchased from sigma company) coating buffer (CBS (1L, pH=9.6): Na
2cO
31.59g, NaHCO
32.93g) according to 1:1000 preparation, then use pipettor to add in microwell plate (one of solid phase carrier 1), every hole 100uL, then at 2 DEG C ~ 8 DEG C, incubation time is more than or equal to 12 hours, then washs 3 times with cleansing solution;
Close: the microwell plate washed is patted dry residual liquid on gauze, and then at 37 DEG C, every hole adds the confining liquid (PBS+0.5%BSA) of 200 μ L, close 2 hours; Directly confining liquid is outwelled, after patting dry Liquid Residue after end; If can not use immediately, be then dry microwell plate under the condition of less than 50% at room epidemic disaster, then seal preservation;
The preparation of 1.2 antigens: adopting Sample dilution (PBS) rubella virus antigen to be diluted to concentration is 0.5 ~ 1mg/mL;
The preparation of 1.3 HRP-monoclonal antibodies 1 and HRP-monoclonal antibody 2: monoclonal antibody 1 and monoclonal antibody 2 (self-control) adopt horseradish peroxidase (to be called for short HRP, be purchased from sigma company) mark according to the glutaraldehyde method of improvement, first dilute according to 1:20000 with Sample dilution before using, re-use;
The preparation of 1.4 substrate solutions and stop buffer:
Substrate A liquid: TMB200mg, absolute ethyl alcohol (or DMSO) 100mL, adds distilled water to 1000mL;
Substrate B liquid: Na
2hPO
414.60g, citric acid 9.33g, 0.75% hydrogen peroxide urea 6.4mL, adds tri-distilled water to 1000mL, regulates pH to 5.0 ~ 5.4;
Stop buffer: the concentrated sulphuric acid measuring 13.6mL98%, then slowly adds in distilled water, and ceaselessly stirs, and is finally settled to 500mL;
The detection method of 1.5 kits:
Band being detected sample uses sample diluting liquid (PBS) according to 1:100 dilution, and then add in the microwell plate prepared, every hole adds 100uL, do 2 critical contrasts simultaneously, at 37 DEG C, hatch 1 hour, then use cleansing solution (PBST) to wash 3 times;
After antigen, HRP-monoclonal antibody 1 and HRP-monoclonal antibody 2 are mixed and made into compound according to 1:1:1, every hole adds 100uL, at 37 DEG C, hatches 0.5 ~ 1 hour, then uses cleansing solution (PBST) to wash 3 times;
Every hole adds substrate A liquid and each mixing of B liquid, and at room temperature lucifuge leaves standstill 15 ~ 30 minutes;
Every hole adds stop buffer 1, in 30 minutes, read its OD value by microplate reader at 450nm wavelength place;
Concrete result criterion is as follows:
Negative: the mean value of OD value < 0.9 × critical control wells;
Positive: the mean value of OD value < 1.1 × critical control wells;
Grey area: OD value is in 1 ± 10% scope of critical control wells mean value.
1.6 testing result
1.6.1 sensitivity for analysis and specificity
Measure 100 routine negative sample through clinical confirmation and 30 routine positive sample by the rubella virus IgM antibody detection kit for preparing, the OD numerical value of reading is as shown in table 1.
The testing result of table 1 kit
Sensitivity for analysis: 30/ (30+0) × 100%=100%
Analyze specificity: 100/ (100+0) × 100%=100%
1.6.2 detection is tested
The kit (buying other companies) of the antigen+HRP-monoclonal antibody 1 that employing rubella virus is prepared separately, antigen+HRP-monoclonal antibody 2, antigen+HRP-monoclonal antibody 1+HRP-monoclonal antibody 2, directly labelled antigen, detect 28 parts of Blood serum plate (WHO), testing result is as shown in table 2.
The testing result of table 2 Blood serum plate (WHO)
As can be seen from the above results, after the present invention adopts 2 to mark monoclonal antibody, detection signal is obviously strengthened, and especially when detecting weak positive sample, shows particularly outstanding, and when detecting negative sample, then basically identical with additive method; As can be seen from the table, market adopts the prize law of direct labelled antigen pattern, in testing process, also occur false positive phenomenon simultaneously.
Described on comprehensive, technical scheme of the present invention can fully effectively complete foregoing invention object, and structural principle of the present invention and the principle of work and power are all verified in an embodiment fully, and effect and the object of expection can be reached, and embodiments of the invention also can according to these principles in the enterprising line translation of different solid phase carriers, be applicable to the antigen of other types too, therefore, the present invention includes all in claim mention all replacement contents in scope.Any equivalence change done in the present patent application the scope of the claims, within the scope of the claims of all genus this case applications.
Claims (6)
1. detect an immunologic detection method for IgM antibody, it is characterized in that: comprise following steps:
The solidification process of step one, anti-IgM, is placed on solid phase carrier by anti-IgM, under the condition of 2 DEG C ~ 8 DEG C, hatch 12 hours and more than, then wash 3 ~ 5 times;
Step 2, IgM to be measured are combined with anti-IgM, are added in the solid phase carrier having had cured anti-IgM by the IgM to be measured as sample, hatch 0.5 ~ 1.0 hour, wash 3 ~ 5 times;
Step 3, antigen, the mark monoclonal antibody IgM to be measured on solid phase carrier is combined, can after first by antigen, monoclonal antibody mix with mark, then adds in the solid phase carrier that washs in step 2, in constant temperature oven, 37 DEG C of constant temperatures hatch 0.5 ~ 1 hour, hatch end, wash 3 ~ 5 times; Or antigen, mark monoclonal antibody are added in the solid phase carrier washed in step 2 successively, namely first added by antigen, and then add mark monoclonal antibody, in constant temperature oven, 37 DEG C of condition constant-temperature incubations 0.5 ~ 1 hour, hatch end, wash 3 ~ 5 times, wait until detection;
Step 4, input, after step 3 completes, according to the difference of the indicant on mark monoclonal antibody, corresponding instrument and equipment carries out its reaction signal detection.
2. a kind of immunologic detection method detecting IgM antibody as claimed in claim 1, it is characterized in that: described antigen, mark monoclonal antibody and IgM's to be measured hatches in process, antigen share can simultaneously in conjunction with the mark monoclonal antibody of 2 and above different binding site, to increase reaction signal.
3. a kind of immunologic detection method detecting IgM antibody as claimed in claim 1, is characterized in that: described solid phase carrier is microwell plate, glass plate or NC film.
4. a kind of immunologic detection method detecting IgM antibody as claimed in claim 1, is characterized in that: described washing process adopts the cleansing solution containing surfactant.
5. a kind of immunologic detection method detecting IgM antibody as claimed in claim 1, is characterized in that: described anti-IgM is goat-anti IgM or mouse-anti IgM.
6. a kind of immunologic detection method detecting IgM antibody as claimed in claim 1, is characterized in that: the indicant in described mark monoclonal antibody is enzyme, collaurum, collargol, electroselenium, latex, radiomaterial, chemiluminescent substance and fluorescent material.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510092986.4A CN104714028B (en) | 2015-03-02 | 2015-03-02 | Immunodetection method for detecting IgM antibody |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510092986.4A CN104714028B (en) | 2015-03-02 | 2015-03-02 | Immunodetection method for detecting IgM antibody |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104714028A true CN104714028A (en) | 2015-06-17 |
| CN104714028B CN104714028B (en) | 2017-01-25 |
Family
ID=53413536
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510092986.4A Active CN104714028B (en) | 2015-03-02 | 2015-03-02 | Immunodetection method for detecting IgM antibody |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104714028B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105785045A (en) * | 2016-04-13 | 2016-07-20 | 柏荣诊断产品（上海）有限公司 | High-performance detection reagent kit for human blood immunoglobulin M |
| CN108169475A (en) * | 2017-12-18 | 2018-06-15 | 郑州安图生物工程股份有限公司 | A kind of Respiratory Syncytial Virus(RSV) IgM antibody detection kit |
| CN116143931A (en) * | 2021-11-20 | 2023-05-23 | 东莞市朋志生物科技有限公司 | Anti-human IgM antibody and preparation method and application thereof |
| WO2023088402A1 (en) * | 2021-11-22 | 2023-05-25 | 菲鹏生物股份有限公司 | Method for detecting antibody by means of capture method and application |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4829012A (en) * | 1985-01-24 | 1989-05-09 | L'association Internationale Abut Scientifique, Dite: "Institut International De Pathologie Cellulaire Et Moleculaire" | Method for immunological assay of antibodies of various types in a liquid sample |
| US20040126904A1 (en) * | 1997-11-18 | 2004-07-01 | Bio-Rad Laboratories, Inc. | Multiplex flow assays preferably with magnetic particles as solid phase |
| CN1902496A (en) * | 2003-11-07 | 2007-01-24 | 赫普金尼克斯股份有限公司 | Binding assay components |
| CN101013133A (en) * | 2007-02-12 | 2007-08-08 | 北京英诺特生物技术有限公司 | Colloidal gold chromatography strip for detecting specific IgM antibody and method for making same |
| CN101711361A (en) * | 2007-06-08 | 2010-05-19 | 博瑞巴斯德公司 | Detect the multiplex method that infects |
-
2015
- 2015-03-02 CN CN201510092986.4A patent/CN104714028B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4829012A (en) * | 1985-01-24 | 1989-05-09 | L'association Internationale Abut Scientifique, Dite: "Institut International De Pathologie Cellulaire Et Moleculaire" | Method for immunological assay of antibodies of various types in a liquid sample |
| US20040126904A1 (en) * | 1997-11-18 | 2004-07-01 | Bio-Rad Laboratories, Inc. | Multiplex flow assays preferably with magnetic particles as solid phase |
| CN1902496A (en) * | 2003-11-07 | 2007-01-24 | 赫普金尼克斯股份有限公司 | Binding assay components |
| CN101013133A (en) * | 2007-02-12 | 2007-08-08 | 北京英诺特生物技术有限公司 | Colloidal gold chromatography strip for detecting specific IgM antibody and method for making same |
| CN101711361A (en) * | 2007-06-08 | 2010-05-19 | 博瑞巴斯德公司 | Detect the multiplex method that infects |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105785045A (en) * | 2016-04-13 | 2016-07-20 | 柏荣诊断产品（上海）有限公司 | High-performance detection reagent kit for human blood immunoglobulin M |
| CN108169475A (en) * | 2017-12-18 | 2018-06-15 | 郑州安图生物工程股份有限公司 | A kind of Respiratory Syncytial Virus(RSV) IgM antibody detection kit |
| CN116143931A (en) * | 2021-11-20 | 2023-05-23 | 东莞市朋志生物科技有限公司 | Anti-human IgM antibody and preparation method and application thereof |
| CN116143931B (en) * | 2021-11-20 | 2023-10-31 | 东莞市朋志生物科技有限公司 | Anti-human IgM antibody and preparation method and application thereof |
| WO2023088402A1 (en) * | 2021-11-22 | 2023-05-25 | 菲鹏生物股份有限公司 | Method for detecting antibody by means of capture method and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104714028B (en) | 2017-01-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105548565B (en) | A kind of kit and its preparation and application for being used to detect schizotrypanum cruzi antibody | |
| CN104614536B (en) | A kind of kit for detecting G17 and its preparation method and application | |
| CN101713779B (en) | Method for performing immunological test on biomolecules by avidin/streptavidin magnetic composite particles | |
| CN104698184B (en) | The test kit of detection sugar antigen | |
| CN104714028A (en) | Immunodetection method for detecting IgM antibody | |
| CN110862881A (en) | Special cleaning solution or diluent for full-automatic chemiluminescence determinator and preparation method thereof | |
| CN101196518A (en) | Hepatitis virus type C immune body chemiluminescence method diagnostic reagent kit and its producing method | |
| CN103837674A (en) | Method for detecting specific IgE antibody, kit used in method and preparation and using methods for kit | |
| CN101377514A (en) | Insulin autoantibody chemiluminescence immune analysis determination reagent kit and preparing method thereof | |
| CN104237520A (en) | Hepatitis c virus antigen-antibody joint detection reagent box and preparation method thereof | |
| CN101178404B (en) | Human immunodeficiency virus antibody chemiluminescence immune analyzing diagnose reagent box and method of producing the same | |
| CN108508001A (en) | Chemiluminescence detection kit | |
| WO2014005527A1 (en) | Method for improving sensitivity of competitive immunoassay | |
| JP7209498B2 (en) | Immunoassay method for hepatitis B virus core antibody | |
| CN102183636A (en) | Diagnostic kit for anti-strep-A DNase B antibody with chemiluminescence immunoassay and using method thereof | |
| CN103513036B (en) | The CLEIA detection kit of salmonella and detection method | |
| CN105758846A (en) | Chemiluminescence enzyme-linked immunosorbent assay reagent kit for detecting clenbuterol | |
| JPH03502248A (en) | Aqueous washing solution for assay, diagnostic test kit and method for measuring herpes simplex virus | |
| KR20140145121A (en) | Immunological analysis method and reagent | |
| CN104849455A (en) | Preparation method and application of protein chip based on serological detection of syphilis | |
| CN102368068A (en) | Kit for detecting chlamydia pneumoniae IgM antibody | |
| CN102095881A (en) | Chemiluminescence quantitative detection kit for free thyroxine | |
| CN105004861A (en) | Human chromogranin A immunoassay method and determination kit | |
| CN109633163A (en) | The two-in-one detection kit of Procalcitonin/c reactive protein | |
| CN105974127A (en) | Human neutrophil apolipoprotein heterodimer quantifying device based on enzyme-linked immune adsorption technology |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20190422 Address after: 518000 6 301A, No. 14, Hangzi Zhongxing Road, Hangzi Street, Pingshan District, Shenzhen City, Guangdong Province Patentee after: Shenzhen Kerunda Bioengineering Co., Ltd. Address before: 518172 Room B1301, 13 floors, Building B, No. 2, Tian'an Digital Innovation Park, Longgang District, Shenzhen, Guangdong Province, at the junction of Qinglin West Road and Huangge North Road, Longgang District, Longcheng Street Center Patentee before: Kai Ruide Bioisystech Co., Ltd of Shenzhen |