CN110568183A - substrate solution for chemiluminescence immunoassay analyzer and preparation method thereof - Google Patents
substrate solution for chemiluminescence immunoassay analyzer and preparation method thereof Download PDFInfo
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- CN110568183A CN110568183A CN201910880472.3A CN201910880472A CN110568183A CN 110568183 A CN110568183 A CN 110568183A CN 201910880472 A CN201910880472 A CN 201910880472A CN 110568183 A CN110568183 A CN 110568183A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
Abstract
The invention discloses a substrate liquid for a chemiluminescence immunoassay analyzer and a preparation method thereof, the substrate liquid is suitable for domestic and foreign full-automatic chemiluminescence immunoassay analyzers such as Yapeh, Kanghua and the like, has low cost, is compared with the Yapeh substrate liquid for on-machine experiments, has higher signal-to-noise ratio and higher luminous intensity, has measured value repeatability superior to that of the Yapeh substrate liquid, and can completely and effectively replace the Yapeh substrate liquid which is widely sold in the market and has high price, thereby effectively solving the problem that domestic hospitals and manufacturers excessively rely on import.
Description
Technical Field
The invention relates to the technical field of chemiluminescence immunoassay.
in particular to a substrate liquid for a chemiluminescence immunoassay analyzer and a preparation method thereof.
background
in the 70 s of the 20 th century, the developed national medical community of Europe and America appeared to detect human diseases by using chemiluminescence immunoassay, which combines chemiluminescence method with immunoreaction and has immunoreaction specificitythe magnetic particle chemiluminescence detection technology is characterized by non-radioactive labeling and no radioactive pollution, is a latest detection technology in the in vitro diagnosis industry, is widely favored by various domestic and foreign clinical laboratories of the great three hospitals, is applied to detection of items such as cardiac markers, tumor markers, thyroid function, metabolism, sex hormones and the like, is popularized and developed to be mature at home and abroad at present, is still in the beginning stage at home, has a wide development prospect, and among the magnetic particle chemiluminescence labeled by acridinium ester is the fuzz of the technology. In the alkaline state H2O2in the solution, when the acridinium ester molecule is attacked by hydrogen peroxide ions, unstable dioxyethane is generated and decomposed into CO2And an electronically excited state of N-methylacridone which emits photons having a maximum emission wavelength of 430nm when it returns to the ground state. The compound is characterized in that the non-luminous substituent part connected to the acridinium ring is separated from the acridinium ring before the electronic excited state intermediate is formed in the luminous reaction, namely, the non-luminous part is separated from the luminous part, so that the luminous efficiency is not influenced by the structure of the substituent group basically. No need of catalyst for chemiluminescence of acridine ester or acridine sulfonamide compound in the presence of H2O2can emit light in the dilute alkaline solution. Therefore, the application of the method to chemiluminescence detection has many advantages. The method has the advantages that the background luminescence is low, and the signal-to-noise ratio is high; secondly, the interference factors of the luminescence reaction are few; the light release is fast and centralized, the luminous efficiency is high, and the luminous intensity is large; fourthly, the protein is easy to be connected and the photon yield is not reduced after the connection; the marker is stable (can be stored for several months at 2-8 ℃). Acridinium esters or acridinium sulfonamides are therefore a very effective, very good class of chemiluminescent labels.
at present, manufacturers at home and abroad are interested in independently researching and developing chemiluminescent substrate liquid and matching with self products. The Yapek substrate liquid is taken as a representative abroad, and domestic manufacturers such as Shenzhen Fenpeng, Suzhou Chang Guang Hua Yi, Shenzhen Yahuilong, Weihaiwei Gao, Xiamen Wantakairui, Nanjing Digranos and the like all develop autonomous substrates, but generally the signal-to-noise ratio and the repeatability are poor, and the Yapek substrate liquid is an internationally recognized better product but is expensive.
Disclosure of Invention
the invention aims to overcome the defects of the traditional technology and provides the substrate liquid for the chemiluminescence immunoassay analyzer and the preparation method thereof, which have the advantages of low cost, excellent signal-to-noise ratio and repeatability and capability of meeting the requirements of the medical field.
The aim of the invention is achieved by the following technical measures:
a substrate liquid for a chemiluminescence immunoassay analyzer comprises a pre-excitation liquid and an excitation liquid;
The pre-excitation liquid comprises the following components:
fluoboric acid: 0.05M-0.2M;
H202：0.05％-0.2％；
EDTA：5mM-10mM；
PC-300：0.05％-0.1％；
the exciting liquid comprises the following components:
NaOH：0.1M-0.35M；
NP-10：0.5％-2.5％；
DMF：0.1％-0.25％；
Krovin600：0.05％-1％。
the fluoboric acid in the pre-excitation liquid is mainly used for adjusting the pH value of the system to ensure that the system has certain acidity, H2O2Has strong oxidizing property and can be used as an oxidizing agent. The pre-excitation liquid mainly plays a role in cracking the marker from the complex and providing an acidic environment to prevent the premature reaction.
NaOH in the excitation liquid mainly enables the solution to have certain ionic strength and certain alkaline environment, and the chemiluminescence reaction is excited.
as an improvement: comprises pre-excitation liquid and excitation liquid;
The pre-excitation liquid comprises the following components:
fluoboric acid: 0.05M;
H202：0.2％；
EDTA：5mM；
PC-300：0.1％；
the exciting liquid comprises the following components:
NaOH：0.1M；
NP-10：2.5％；
DMF：0.1％；
Krovin600：1％。
as an improvement: comprises pre-excitation liquid and excitation liquid;
the pre-excitation liquid comprises the following components:
fluoboric acid: 0.2M;
H202：0.05％；
EDTA：10mM；
PC-300：0.05％；
the exciting liquid comprises the following components:
NaOH：0.35M；
NP-10：0.5％；
DMF：0.25％；
Krovin600：0.05％。
as an improvement: comprises pre-excitation liquid and excitation liquid;
the pre-excitation liquid comprises the following components:
Fluoboric acid: 0.125M;
H202：0.125％；
EDTA：7.5mM；
PC-300：0.075％；
the exciting liquid comprises the following components:
NaOH：0.225M；
NP-10：1.5％；
DMF：0.175％；
Krovin600：0.075％。
a method for preparing substrate liquid for a chemiluminescence immunoassay analyzer;
the preparation method of the pre-excitation liquid comprises the following steps:
A1, taking 800ml of pure water;
a2, adding 4.97ml to 19.88ml of fluoboric acid;
A3, adding 1.66ml-6.64ml H2O2；
a4, adding 1.69g-3.38g of EDTA;
a5, adding 0.5ml-1ml of PC-300;
a6, stirring thoroughly, mixing well, adjusting pH to 1.50 + -0.2 with 1M NaOH/1M HCl;
a7, and the volume is adjusted to 1L by pure water.
the preparation method of the excitation liquid comprises the following steps:
b1, taking 800ml of pure water;
b2, adding 4g-14g of NaOH;
b3, adding 5ml-25ml NP-10;
B4, adding 1ml-2.5ml DMF;
b5, adding 0.5ml-1ml Krovin 600;
B6, stirring fully and mixing uniformly, and adjusting the pH value to be 12.50 +/-0.2 by using 1MNaOH/1 MHCl;
B7, using pure water to fix the volume to 1L.
As an improvement:
the preparation method of the pre-excitation liquid comprises the following steps:
a1, taking 800ml of pure water;
a2, adding 4.97ml of fluoboric acid;
a3, 6.64ml H2O2；
a4, adding 1.69g of EDTA;
a5, adding 0.5ml of PC-300;
a6, stirring thoroughly, mixing well, adjusting pH to 1.50 + -0.2 with 1M NaOH/1M HCl;
a7, and the volume is adjusted to 1L by pure water.
the preparation method of the excitation liquid comprises the following steps:
B1, taking 800ml of pure water;
b2, adding 4g of NaOH;
B3, adding 25ml of NP-10;
B4, adding 1 mlmlmlDMF;
b5, adding 1ml of Krovin 600;
b6, stirring fully and mixing uniformly, and adjusting the pH value to be 12.50 +/-0.2 by using 1MNaOH/1 MHCl;
b7, using pure water to fix the volume to 1L.
as an improvement:
the preparation method of the pre-excitation liquid comprises the following steps:
a1, taking 800ml of pure water;
A2, adding 19.88ml of fluoboric acid;
a3, addition of 1.66ml H2O2；
a4, adding 3.38g of EDTA;
a5, adding 0.5ml of PC-300;
A6, stirring thoroughly, mixing well, adjusting pH to 1.50 + -0.2 with 1M NaOH/1M HCl;
a7, and the volume is adjusted to 1L by pure water.
The preparation method of the excitation liquid comprises the following steps:
b1, taking 800ml of pure water;
b2, adding 14g of NaOH;
b3, adding 5ml of NP-10;
b4, adding 1 mlmlmlDMF;
B5, adding 0.5ml of Krovin 600;
b6, stirring fully and mixing uniformly, and adjusting the pH value to be 12.50 +/-0.2 by using 1MNaOH/1 MHCl;
b7, using pure water to fix the volume to 1L.
as an improvement:
the preparation method of the pre-excitation liquid comprises the following steps:
a1, taking 800ml of pure water;
A2, adding 12.43ml of fluoboric acid;
a3, addition of 4.15ml H2O2；
A4, adding 2.54g of EDTA;
a5, adding 0.75ml of PC-300;
a6, stirring thoroughly, mixing well, adjusting pH to 1.50 + -0.2 with 1M NaOH/1M HCl;
a7, and the volume is adjusted to 1L by pure water.
The preparation method of the excitation liquid comprises the following steps:
b1, taking 800ml of pure water;
b2, adding 9g of NaOH;
b3, adding 15ml of NP-10;
b4, adding 1.75ml of DMF;
b5, adding 0.75ml of Krovin 600;
b6, stirring fully and mixing uniformly, and adjusting the pH value to be 12.50 +/-0.2 by using 1MNaOH/1 MHCl;
b7, using pure water to fix the volume to 1L.
due to the adoption of the technical scheme, compared with the prior art, the invention has the advantages that:
1. low cost, and is suitable for Yapeh, Kanghua and other domestic and foreign full-automatic chemiluminescence immune analyzers.
2. Compared with Yapeh substrate liquid, the technical scheme has higher signal-to-noise ratio, higher luminous intensity and better measured value repeatability than that of the Yapeh substrate liquid, and can completely and effectively replace the Yapeh substrate liquid which is widely sold in the market and has high price, thereby effectively solving the problem that domestic hospitals and manufacturers excessively rely on import.
Detailed Description
example 1: a substrate liquid for chemiluminescence immunoassay analyzer comprises a pre-excitation liquid and an excitation liquid.
the pre-excitation liquid comprises the following components:
fluoboric acid: 0.05M;
H202：0.2％；
EDTA：5mM；
PC-300：0.1％；
the exciting liquid comprises the following components:
NaOH：0.1M；
NP-10：2.5％；
DMF：0.1％；
Krovin600：1％。
example 2: a substrate liquid for chemiluminescence immunoassay analyzer comprises a pre-excitation liquid and an excitation liquid.
The pre-excitation liquid comprises the following components:
fluoboric acid: 0.2M;
H202：0.05％；
EDTA：10mM；
PC-300：0.05％；
the exciting liquid comprises the following components:
NaOH：0.35M；
NP-10：0.5％；
DMF：0.25％；
Krovin600：0.05％。
example 3: a substrate liquid for chemiluminescence immunoassay analyzer comprises a pre-excitation liquid and an excitation liquid.
The pre-excitation liquid comprises the following components:
Fluoboric acid: 0.125M;
H202：0.125％；
EDTA：7.5mM；
PC-300：0.075％；
the exciting liquid comprises the following components:
NaOH：0.225M；
NP-10：1.5％；
DMF：0.175％；
Krovin600：0.075％。
example 4: the method for preparing a substrate solution for a chemiluminescent immunoassay analyzer as described in example 1.
the preparation method of the pre-excitation liquid comprises the following steps:
a1, measuring 800ml of pure water by using a measuring cylinder, and adding into a container;
a2, measuring 4.97ml of fluoboric acid by a pipette, and adding the fluoboric acid into a container;
A3, measuring 6.64mlH by using a pipette202adding into a container;
a4, weighing 1.69g of EDTA by using an electronic balance, and adding into a container;
A5, measuring 0.5ml of PC-300 by a pipette, and adding into a container;
a6, stirring thoroughly, mixing well, adjusting pH to 1.50 + -0.2 with 1M NaOH/1M HCl;
A7, and the volume is adjusted to 1L by pure water.
the preparation method of the excitation liquid comprises the following steps:
B1, measuring 800ml of pure water by using a measuring cylinder, and adding into a container;
b2, weighing 4g of NaOH by using an electronic balance, and adding the NaOH into a container;
b3, measuring 25ml NP-10 by using a pipette, and adding into a container;
b4, measuring 1 mlmlmlmlDMF by using a pipette, and adding the obtained product into a container;
B5, measuring 1ml of Krovin600 by using a pipette, and adding the Krovin600 into a container;
B6, stirring fully and mixing uniformly, and adjusting the pH value to be 12.50 +/-0.2 by using 1MNaOH/1 MHCl;
b7, using pure water to fix the volume to 1L.
example 5: the method for preparing a substrate solution for a chemiluminescent immunoassay analyzer as described in example 2.
the preparation method of the pre-excitation liquid comprises the following steps:
a1, measuring 800ml of pure water by using a measuring cylinder, and adding into a container;
a2, measuring 19.88ml of fluoboric acid by a pipette, and adding the fluoboric acid into a container;
a3, measuring 1.66mlH by using a pipette202adding into a container;
a4, weighing 3.38g of EDTA by using an electronic balance, and adding into a container;
a5, measuring 0.5ml of PC-300 by a pipette, and adding into a container;
A6, stirring thoroughly, mixing well, adjusting pH to 1.50 + -0.2 with 1M NaOH/1M HCl;
a7, and the volume is adjusted to 1L by pure water.
the preparation method of the excitation liquid comprises the following steps:
b1, measuring 800ml of pure water by using a measuring cylinder, and adding into a container;
b2, weighing 14g of NaOH by using an electronic balance, and adding the NaOH into a container;
b3, measuring 5ml NP-10 by using a pipette, and adding into a container;
b4, measuring 1 mlmlmlmlDMF by using a pipette, and adding the obtained product into a container;
B5, measuring 0.5ml of Krovin600 by using a pipette, and adding the Krovin600 into a container;
b6, stirring fully and mixing uniformly, and adjusting the pH value to be 12.50 +/-0.2 by using 1MNaOH/1 MHCl;
B7, using pure water to fix the volume to 1L.
Example 6: the method for preparing a substrate solution for a chemiluminescent immunoassay analyzer as described in example 3.
The preparation method of the pre-excitation liquid comprises the following steps:
A1, measuring 800ml of pure water by using a measuring cylinder, and adding into a container;
A2, measuring 12.43ml of fluoroboric acid by using a pipette, and adding the fluoroboric acid into a container;
a3, measuring 4.15mlH by using a pipette202adding into a container;
A4, weighing 2.54g of EDTA by using an electronic balance, and adding into a container;
a5, measuring 0.75ml of PC-300 by a pipette, and adding into a container;
a6, stirring thoroughly, mixing well, adjusting pH to 1.50 + -0.2 with 1M NaOH/1M HCl;
a7, and the volume is adjusted to 1L by pure water.
the preparation method of the excitation liquid comprises the following steps:
b1, measuring 800ml of pure water by using a measuring cylinder, and adding into a container;
b2, weighing 9g of NaOH by using an electronic balance, and adding the NaOH into a container;
b3, measuring 15ml NP-10 by using a pipette, and adding into a container;
b4, measuring 1.75ml of DMF by using a pipette, and adding the DMF into a container;
b5, measuring 0.75ml of Krovin600 by using a pipette, and adding the Krovin600 into a container;
B6, stirring fully and mixing uniformly, and adjusting the pH value to be 12.50 +/-0.2 by using 1MNaOH/1 MHCl;
B7, using pure water to fix the volume to 1L.
the computer experiment was performed as described in example 3, and the raw data are shown below.
Table one: and (5) verifying the repeatability and the luminous intensity of the cleaning magnetic beads.
table two: and (5) verifying the luminous intensity of the TSH item.
table three: and (5) verifying the repeatability of the TSH project.
Table four: PGII project luminous intensity verification
table five: and (5) verifying the repeatability of the PGII project.
the data comparison is shown below.
table one: magnetic bead washing verification (low, medium, high 3 concentration).
table two: TSH project
table three: PGII project
although several embodiments of the present invention have been described in detail, the description is only for the preferred embodiments of the present invention and should not be construed as limiting the scope of the present invention. All equivalent changes and modifications made within the scope of the present invention shall fall within the scope of the present invention.
Claims (8)
1. a substrate liquid for a chemiluminescence immunoassay analyzer is characterized in that: comprises pre-excitation liquid and excitation liquid;
the pre-excitation liquid comprises the following components:
fluoboric acid: 0.05M-0.2M;
H202：0.05%-0.2%；
EDTA：5mM-10mM；
PC-300：0.05%-0.1%；
the exciting liquid comprises the following components:
NaOH：0.1M-0.35M；
NP-10：0.5%-2.5%；
DMF：0.1%-0.25%；
Krovin600：0.05%-1%。
2. the substrate liquid for a chemiluminescent immunoassay analyzer according to claim 1, wherein: comprises pre-excitation liquid and excitation liquid;
the pre-excitation liquid comprises the following components:
fluoboric acid: 0.05M;
H202：0.2%；
EDTA：5mM；
PC-300：0.1%；
the exciting liquid comprises the following components:
NaOH：0.1M；
NP-10：2.5%；
DMF：0.1%；
Krovin600：1%。
3. The substrate liquid for a chemiluminescent immunoassay analyzer according to claim 1, wherein: comprises pre-excitation liquid and excitation liquid;
the pre-excitation liquid comprises the following components:
fluoboric acid: 0.2M;
H202：0.05%；
EDTA：10mM；
PC-300：0.05%；
The exciting liquid comprises the following components:
NaOH：0.35M；
NP-10：0.5%；
DMF：0.25%；
Krovin600：0.05%。
4. The substrate liquid for a chemiluminescent immunoassay analyzer according to claim 1, wherein: comprises pre-excitation liquid and excitation liquid;
the pre-excitation liquid comprises the following components:
fluoboric acid: 0.125M;
H202：0.125%；
EDTA：7.5mM；
PC-300：0.075%；
the exciting liquid comprises the following components:
NaOH：0.225M；
NP-10：1.5%；
DMF：0.175%；
Krovin600：0.075%。
5. a preparation method of substrate liquid for a chemiluminescence immunoassay analyzer is characterized by comprising the following steps:
the preparation method of the pre-excitation liquid comprises the following steps:
a1, taking 800ml of pure water;
a2, adding 4.97ml to 19.88ml of fluoboric acid;
A3, adding 1.66ml-6.64ml H2O2；
a4, adding 1.69g-3.38g of EDTA;
a5, adding 0.5ml-1ml of PC-300;
a6, stirring thoroughly, mixing well, adjusting pH to 1.50 + -0.2 with 1M NaOH/1M HCl;
a7, adding pure water to a constant volume of 1L;
the preparation method of the excitation liquid comprises the following steps:
b1, taking 800ml of pure water;
b2, adding 4g-14g of NaOH;
b3, adding 5ml-25ml NP-10;
b4, adding 1ml-2.5ml DMF;
b5, adding 0.5ml-1ml Krovin 600;
b6, stirring fully and mixing uniformly, and adjusting the pH value to be 12.50 +/-0.2 by using 1MNaOH/1 MHCl;
b7, using pure water to fix the volume to 1L.
6. the method for preparing a substrate liquid for a chemiluminescent immunoassay analyzer according to claim 5, wherein:
the preparation method of the pre-excitation liquid comprises the following steps:
A1, taking 800ml of pure water;
A2, adding 4.97ml of fluoboric acid;
a3, 6.64ml H2O2；
a4, adding 1.69g of EDTA;
a5, adding 0.5ml of PC-300;
a6, stirring thoroughly, mixing well, adjusting pH to 1.50 + -0.2 with 1M NaOH/1M HCl;
a7, adding pure water to a constant volume of 1L;
The preparation method of the excitation liquid comprises the following steps:
b1, taking 800ml of pure water;
B2, adding 4g of NaOH;
B3, adding 25ml of NP-10;
B4, adding 1 mlmlmlDMF;
B5, adding 1ml of Krovin 600;
b6, stirring fully and mixing uniformly, and adjusting the pH value to be 12.50 +/-0.2 by using 1MNaOH/1 MHCl;
b7, using pure water to fix the volume to 1L.
7. the method for preparing a substrate liquid for a chemiluminescent immunoassay analyzer according to claim 5, wherein:
The preparation method of the pre-excitation liquid comprises the following steps:
a1, taking 800ml of pure water;
a2, adding 19.88ml of fluoboric acid;
A3, addition of 1.66ml H2O2；
a4, adding 3.38g of EDTA;
a5, adding 0.5ml of PC-300;
a6, stirring thoroughly, mixing well, adjusting pH to 1.50 + -0.2 with 1M NaOH/1M HCl;
a7, adding pure water to a constant volume of 1L;
the preparation method of the excitation liquid comprises the following steps:
b1, taking 800ml of pure water;
B2, adding 14g of NaOH;
b3, adding 5ml of NP-10;
b4, adding 1 mlmlmlDMF;
b5, adding 0.5ml of Krovin 600;
b6, stirring fully and mixing uniformly, and adjusting the pH value to be 12.50 +/-0.2 by using 1MNaOH/1 MHCl;
B7, using pure water to fix the volume to 1L.
8. the method for preparing a substrate liquid for a chemiluminescent immunoassay analyzer according to claim 5, wherein:
the preparation method of the pre-excitation liquid comprises the following steps:
a1, taking 800ml of pure water;
a2, adding 12.43ml of fluoboric acid;
A3, addition of 4.15ml H2O2；
a4, adding 2.54g of EDTA;
a5, adding 0.75ml of PC-300;
A6, stirring thoroughly, mixing well, adjusting pH to 1.50 + -0.2 with 1M NaOH/1M HCl;
a7, adding pure water to a constant volume of 1L;
The preparation method of the excitation liquid comprises the following steps:
b1, taking 800ml of pure water;
b2, adding 9g of NaOH;
b3, adding 15ml of NP-10;
b4, adding 1.75ml of DMF;
b5, adding 0.75ml of Krovin 600;
B6, stirring fully and mixing uniformly, and adjusting the pH value to be 12.50 +/-0.2 by using 1MNaOH/1 MHCl;
b7, using pure water to fix the volume to 1L.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117129668A (en) * | 2023-10-27 | 2023-11-28 | 江西赛基生物技术有限公司 | Cleaning solution for chemiluminescence immunoassay and preparation method and application thereof |
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| CN117129668B (en) * | 2023-10-27 | 2024-01-09 | 江西赛基生物技术有限公司 | Cleaning solution for chemiluminescence immunoassay and preparation method and application thereof |
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Application publication date: 20191213 |