CN115363637A - Saliva sample acquisition card and method for acquiring saliva sample by using same - Google Patents
Saliva sample acquisition card and method for acquiring saliva sample by using same Download PDFInfo
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- CN115363637A CN115363637A CN202211010799.3A CN202211010799A CN115363637A CN 115363637 A CN115363637 A CN 115363637A CN 202211010799 A CN202211010799 A CN 202211010799A CN 115363637 A CN115363637 A CN 115363637A
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- 210000003296 saliva Anatomy 0.000 title claims abstract description 53
- 238000000034 method Methods 0.000 title claims abstract description 14
- 239000007788 liquid Substances 0.000 claims abstract description 16
- 238000005070 sampling Methods 0.000 claims abstract description 14
- 239000000243 solution Substances 0.000 claims description 11
- 238000004321 preservation Methods 0.000 claims description 10
- 239000003761 preservation solution Substances 0.000 claims description 10
- 239000002994 raw material Substances 0.000 claims description 6
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 5
- 238000001514 detection method Methods 0.000 claims description 5
- 239000012530 fluid Substances 0.000 claims description 5
- 239000007983 Tris buffer Substances 0.000 claims description 4
- ZJYYHGLJYGJLLN-UHFFFAOYSA-N guanidinium thiocyanate Chemical compound SC#N.NC(N)=N ZJYYHGLJYGJLLN-UHFFFAOYSA-N 0.000 claims description 4
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 4
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 3
- 229920004890 Triton X-100 Polymers 0.000 claims description 3
- 239000003755 preservative agent Substances 0.000 claims 1
- 230000002335 preservative effect Effects 0.000 claims 1
- 241000894006 Bacteria Species 0.000 abstract description 8
- 241000700605 Viruses Species 0.000 abstract description 8
- 230000000415 inactivating effect Effects 0.000 abstract description 4
- 238000010525 oxidative degradation reaction Methods 0.000 abstract description 3
- 230000003313 weakening effect Effects 0.000 abstract description 3
- 238000002558 medical inspection Methods 0.000 abstract description 2
- 239000000523 sample Substances 0.000 description 32
- 239000001963 growth medium Substances 0.000 description 20
- 230000003385 bacteriostatic effect Effects 0.000 description 9
- 230000000052 comparative effect Effects 0.000 description 9
- 230000000694 effects Effects 0.000 description 4
- 239000000463 material Substances 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 239000007858 starting material Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 229920000742 Cotton Polymers 0.000 description 1
- 241000588914 Enterobacter Species 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000008139 complexing agent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000011840 criminal investigation Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- YQOKLYTXVFAUCW-UHFFFAOYSA-N guanidine;isothiocyanic acid Chemical compound N=C=S.NC(N)=N YQOKLYTXVFAUCW-UHFFFAOYSA-N 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 210000002200 mouth mucosa Anatomy 0.000 description 1
- 239000006179 pH buffering agent Substances 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B10/00—Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
- A61B10/0045—Devices for taking samples of body liquids
- A61B10/0051—Devices for taking samples of body liquids for taking saliva or sputum samples
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pathology (AREA)
- Hematology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Biophysics (AREA)
- Surgery (AREA)
- Biotechnology (AREA)
- Animal Behavior & Ethology (AREA)
- Microbiology (AREA)
- Pulmonology (AREA)
- Medical Informatics (AREA)
- Heart & Thoracic Surgery (AREA)
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- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention belongs to the technical field of biological and medical inspection, and relates to a saliva sample acquisition card and a method for acquiring a saliva sample by using the same. The invention constructs a saliva sample collection combination comprising a sampling swab, a filter paper card and a sample collection liquid, wherein the sampling swab has a more efficient DNA enrichment capacity, the filter paper card has an indication function and a capacity of prolonging the DNA storage time, and the sample collection liquid has a capacity of weakening DNA oxidative degradation and inactivating viruses and bacteria, so that the risk of contacting the viruses and the bacteria by personnel is reduced.
Description
Technical Field
The invention belongs to the technical field of biological and medical inspection, and relates to a saliva sample acquisition card and a method for acquiring a saliva sample by using the same.
Background
Saliva is an important test material type in forensic science DNA detection work, the essence of the saliva is to detect DNA in oral mucosa exfoliative cells contained in the saliva, and the saliva can be applied to the fields of criminal investigation, health examination, academic research and the like. Saliva is in a liquid form and is not easy to store and transfer, and a saliva sample is fixed in a saliva collecting card form and is widely accepted and applied in the field of DNA detection in forensic science.
Two key problems are encountered when saliva samples are collected by forensic DNA field examination materials: 1. the conventional method of using a cotton swab to collect and transfer the DNA onto a card has poor preservation effect; 2. when unknown on-site test materials possibly attached with viruses, bacteria and the like are sampled, the safety of sampling, transferring and detecting personnel is ensured.
Disclosure of Invention
The invention aims to provide a saliva sample collection card with high-efficiency DNA enrichment capacity and prolonged DNA storage time aiming at the problems in the prior art.
The purpose of the invention can be realized by the following technical scheme:
a saliva sample collecting card comprises a saliva collecting area, a sample personnel information filling area and a sampling information filling and pasting area, and is characterized in that the saliva collecting area contains a collecting liquid, and the collecting liquid comprises the following raw materials: 55-60g/L Tris, 85-95g/L SDS, 0.03-0.08M EDTA, 7-8g/L Na 2 SO 3 8-12g/L litmus.
Tris is used as a PH buffering agent, SDS is used as a complexing agent, the enrichment capacity of DNA can be improved, EDTA is an antibacterial agent, pathogenic microorganisms possibly carried in a biological sample can be inactivated, the safety of sampling personnel can be ensured, litmus is used as a color indicator, and the DNA is ensured to be collected on a collection card.
In the saliva sample collection card, the pH of the collection liquid is 3-5.
The invention also provides a method for collecting saliva samples, which collects the saliva samples by using the collection card
In the above method for collecting a saliva sample, the method comprises the steps of: coating the collected saliva in a saliva collecting area, uniformly spraying a sample preservation solution in the saliva collecting area, fixing a collecting card, and carrying out sample detection.
In the method for collecting a saliva sample, the sample preservation solution comprises the following raw materials: 2-3g/L Triton X100, 80-90g/L guanidinium isothiocyanate and 0.01M Tris-HCl solution.
The invention destroys protein by adding 80-90g/L guanidinium isothiocyanate, has the function of inactivating virus, and can greatly improve the safety of sampling personnel.
In one of the above methods for collecting a saliva sample, the pH of the sample preservation solution is 6.5 to 7.5.
Compared with the prior art, the invention has the following beneficial effects:
the invention constructs a saliva sample collection combination comprising a sampling swab, a filter paper card and a sample collection liquid, wherein the sampling swab has a more efficient DNA enrichment capacity, the filter paper card has an indication function and a capacity of prolonging the DNA storage time, and the sample collection liquid has a capacity of weakening DNA oxidative degradation and inactivating viruses and bacteria, so that the risk of contacting the viruses and the bacteria by personnel is reduced.
Drawings
FIG. 1 is a schematic diagram of a sample acquisition card according to embodiment 1; 1. a saliva collection region; 2. a sample staff information filling area; 3. filling and pasting a sampling information area; 4. and (6) protecting paper.
FIG. 2 is a photograph of the collection card of example 1 placed in a culture medium and cultured for 1 day.
FIG. 3 is a photograph of the collection card of example 2 placed in a culture medium and cultured for 1 day.
FIG. 4 is a photograph of the collection card of example 3 placed in a culture medium and cultured for 1 day.
FIG. 5 is a photograph of the collection card of example 4 placed in a culture medium and cultured for 1 day.
FIG. 6 is a photograph of the collection card of example 5 placed in a culture medium for 1 day.
FIG. 7 is a photograph of the collection card of example 6 placed in a culture medium and cultured for 1 day.
FIG. 8 is a photograph of the collection card of example 7 placed in a culture medium and cultured for 1 day.
FIG. 9 is a photograph of the collection card of comparative example 1 placed in a culture medium and cultured for 1 day.
FIG. 10 is a photograph of the sample card of comparative example 2 placed in a culture medium and cultured for 1 day.
Detailed Description
The following are specific examples of the present invention and further describe the technical solutions of the present invention, but the present invention is not limited to these examples.
Example 1:
a saliva sample collection card as shown in fig. 1, comprising a saliva display collection area, a sample staff information filling area, a sampling information filling and pasting area, wherein the saliva collection area contains a collection liquid, the collection liquid has a pH of 4, and the collection card comprises the following raw materials: 58.2g/L Tris, 90g/LSDS, 0.05M EDTA, 7.6g/L Na 2 SO 3 10g/L litmus.
The collected saliva is coated on a saliva collecting area of a saliva sample collecting card, then the sample preserving fluid is evenly sprayed in the saliva collecting area, and then the collecting card is fixed and sent for detection.
The pH value of the sample preservation solution is 7, and the sample preservation solution comprises the following raw materials: 2.53g/L Triton X100, 86.53g/L guanidinium isothiocyanate, 0.01M Tris-HCl solution.
Example 2:
the difference from example 1 is only that guanidine isothiocyanate was not added to the specimen preservation solution.
Example 3:
the only difference from example 1 is that the saliva collection area is not sprayed with a sample preservation solution.
Example 4:
the difference from example 1 is only that the sample preservation solution has a pH of 6.
Example 5:
the difference from example 1 is only that the sample preservation solution has a pH of 8.
Example 6:
the difference from example 1 was only that the pH of the collected solution was 2.
Example 7:
the only difference from example 1 is that the pH of the collection solution was 6.
Comparative example 1:
the difference from example 1 was only that SDS was not added to the starting material of the collected liquid.
Comparative example 2:
the difference from example 1 was only that EDTA was not added to the starting material of the collected solution.
Respectively dripping enterobacter competent bacteria liquid into saliva collecting areas of the collecting cards prepared in examples 1-7 and comparative examples 1-2, then adding a sample preserving fluid, and finally placing the sample preserving fluid in a culture medium to culture for 1 day to observe the culture medium.
FIG. 2 is a photograph of the collection card of example 1 placed in a culture medium for 1 day, and it can be seen that the collection card of example 1 has bacteriostatic effect and is obvious in effect.
FIG. 3 is a photograph of the collection card of example 2 placed in a culture medium for 1 day, from which it can be seen that the collection card of example 2 has no bacteriostatic effect.
FIG. 4 is a photograph of the collection card of example 3 placed in a culture medium for 1 day, from which it can be seen that the collection card of example 3 has no bacteriostatic effect.
FIG. 5 is a photograph of the collection card of example 4 placed in a culture medium for 1 day, from which it can be seen that the collection card of example 4 has no bacteriostatic effect.
FIG. 6 is a photograph of the collection card of example 5 placed in a culture medium and cultured for 1 day, and it can be seen that the collection card of example 5 has no bacteriostatic effect.
FIG. 7 is a photograph of the collection card of example 6 placed in a culture medium for 1 day, from which it can be seen that the collection card of example 6 has bacteriostatic effect, but the effect is not significant.
FIG. 8 is a photograph of the collection card of example 7 placed in a culture medium and cultured for 1 day, and it can be seen that the collection card of example 7 has bacteriostatic effect, but the effect is not obvious.
FIG. 9 is a photograph of the sample card of comparative example 1 placed in a culture medium and cultured for 1 day, and it can be seen that the sample card of comparative example 1 has no bacteriostatic effect.
FIG. 10 is a photograph of the sample card of comparative example 2 placed in a culture medium and cultured for 1 day, and it can be seen that the sample card of comparative example 2 has no bacteriostatic effect.
In conclusion, the saliva sample collection combination comprising the sampling swab, the filter paper card and the sample collection liquid is constructed, the sampling swab has a high-efficiency DNA enrichment capacity, the filter paper card has an indication function and a capacity of prolonging the DNA storage time, and the sample collection liquid has a capacity of weakening DNA oxidative degradation and inactivating viruses and bacteria, so that the risk of contacting the viruses and the bacteria by personnel is reduced.
The technical scope of the invention claimed by the embodiments of the present application is not exhaustive, and new technical solutions formed by equivalent replacement of single or multiple technical features in the technical solutions of the embodiments are also within the scope of the invention claimed by the present application; in all the embodiments of the present invention, which are listed or not listed, each parameter in the same embodiment only represents an example (i.e., a feasible embodiment) of the technical solution, and there is no strict matching and limiting relationship between the parameters, wherein the parameters may be replaced with each other without departing from the axiom and the requirements of the present invention, unless otherwise specified.
The technical means disclosed by the scheme of the invention are not limited to the technical means disclosed by the technical means, and the technical scheme also comprises the technical scheme formed by any combination of the technical characteristics. While the foregoing is directed to embodiments of the present invention, it will be appreciated by those skilled in the art that various changes and modifications may be made without departing from the principles of the invention, and it is intended that all such changes and modifications be considered as within the scope of the invention.
The specific embodiments described herein are merely illustrative of the spirit of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit or ambit of the invention as defined in the appended claims.
Claims (6)
1. A saliva sample collection card comprises a saliva collection area, a sample personnel information filling area and a sampling information filling and pasting area, and is characterized in that the saliva collection area contains a collection liquid, and the collection liquid comprises the following raw materials: 55-60g/L Tris, 85-95g/L SDS, 0.03-0.08M EDTA, 7-8g/L Na 2 SO 3 8-12g/L litmus.
2. The saliva sample collection card of claim 1, wherein the pH of the collection solution is 3-5.
3. A method for collecting a saliva sample, wherein the saliva sample is collected using the collection card of claim 1.
4. A method of collecting a saliva sample according to claim 3, characterized in that the method comprises the steps of: the collected saliva is coated on a saliva collecting area, the sample preserving fluid is evenly sprayed in the saliva collecting area, and then the collecting card is fixed and sent for detection.
5. The method of claim 4, wherein the sample preservative fluid comprises the following raw materials: 2-3g/L Triton X100, 80-90g/L guanidinium isothiocyanate and 0.01M Tris-HCl solution.
6. A method of sampling saliva according to claim 4 or 5 characterised in that the pH of the sample preservation solution is 6.5-7.5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211010799.3A CN115363637A (en) | 2022-08-23 | 2022-08-23 | Saliva sample acquisition card and method for acquiring saliva sample by using same |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211010799.3A CN115363637A (en) | 2022-08-23 | 2022-08-23 | Saliva sample acquisition card and method for acquiring saliva sample by using same |
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| CN115363637A true CN115363637A (en) | 2022-11-22 |
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| CN202211010799.3A Pending CN115363637A (en) | 2022-08-23 | 2022-08-23 | Saliva sample acquisition card and method for acquiring saliva sample by using same |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103614371A (en) * | 2013-11-21 | 2014-03-05 | 安徽华卫集团禽业有限公司 | Method of simultaneously extracting animal DNA (Deoxyribonucleic Acid) virus and RNA (Ribonucleic Acid) virus nucleic acid in blood serum and double swabs |
| CN104651524A (en) * | 2015-03-13 | 2015-05-27 | 苏州新海生物科技有限公司 | Method for storing biological samples and kit |
| CN107227306A (en) * | 2017-06-26 | 2017-10-03 | 郑州安图生物工程股份有限公司 | A kind of swab eluent with Sample preservation and inactivation function |
| CN207979706U (en) * | 2017-06-19 | 2018-10-19 | 苏州阅微基因技术有限公司 | A kind of saliva capture card |
| CN111902545A (en) * | 2019-01-03 | 2020-11-06 | 杭州诺辉健康科技有限公司 | Composition and method for urine sample preservation and DNA extraction |
-
2022
- 2022-08-23 CN CN202211010799.3A patent/CN115363637A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103614371A (en) * | 2013-11-21 | 2014-03-05 | 安徽华卫集团禽业有限公司 | Method of simultaneously extracting animal DNA (Deoxyribonucleic Acid) virus and RNA (Ribonucleic Acid) virus nucleic acid in blood serum and double swabs |
| CN104651524A (en) * | 2015-03-13 | 2015-05-27 | 苏州新海生物科技有限公司 | Method for storing biological samples and kit |
| CN207979706U (en) * | 2017-06-19 | 2018-10-19 | 苏州阅微基因技术有限公司 | A kind of saliva capture card |
| CN107227306A (en) * | 2017-06-26 | 2017-10-03 | 郑州安图生物工程股份有限公司 | A kind of swab eluent with Sample preservation and inactivation function |
| CN111902545A (en) * | 2019-01-03 | 2020-11-06 | 杭州诺辉健康科技有限公司 | Composition and method for urine sample preservation and DNA extraction |
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