CN106124753A - Electrochemiluminescence buffer and cleanout fluid - Google Patents
Electrochemiluminescence buffer and cleanout fluid Download PDFInfo
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- CN106124753A CN106124753A CN201610538476.XA CN201610538476A CN106124753A CN 106124753 A CN106124753 A CN 106124753A CN 201610538476 A CN201610538476 A CN 201610538476A CN 106124753 A CN106124753 A CN 106124753A
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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
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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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Abstract
The present invention relates to a kind of electrochemiluminescence buffer being applicable to Roche automatic lmunoassays analyzer and cleanout fluid, buffer formulation greatly reduces the consumption of the bigger tripropyl amine (TPA) of toxicity, and add the material of enhanced chemiluminescence, decrease the toxicity of original-pack reagent, reduce the operational risk of docimaster, preparation of reagents is simple, and result is accurate；Cleanout fluid formula increases the composition such as coagula and fibrin effectively cleaned composition, and add Proclin 300 preservative, safeguard electrode further.Cleanout fluid cleaning performance is good, and the holding time is long.The present invention has extremely strong industrial application value.
Description
Technical field
The present invention relates to can be used for chemiluminescent buffer and the cleanout fluid of Roche automatic lmunoassays analyzer, belong to medical treatment
Apparatus external diagnosis reagent field.
Background technology
Electrogenerated chemiluminescence (ECL), also referred to as electrochemiluminescence, is a kind of novelty of combining with electrochemistry of chemiluminescence
Research method.It is to be electrochemically reacted by electrode is applied certain voltage, certain between product or in product and system
Planting and chemical reaction occurs between component, make electron transition arrive excited state and produce excited state material, excited state material returns to ground state and produces
Hair growth promoting light；Or luminescent substance utilizes electrode to provide energy directly to carry out redox reaction, generate unstable intermediate state
Material, intermediate state material can decompose rapidly generation light radiation.
Roche company of Switzerland Cobas e411/e601 Electrogenerated chemiluminescent immunoassay system, is the electrochemistry of application advanced person
The Full-automatic desk type analyser of luminous (ECL) technology.Thyroid, hormone, myocardial damage and tumor-marker, infection it are applied to
Disease, anemia, bone metabolism etc. measure project widely.Its electrochemiluminescsystem system is tris (bipyridine) ruthenium ([Ru (bpy) 32+])-three
Propylamine (TPrA) system, its matching used Procell and Cleancell large usage quantity, supporting original-pack reagent is particularly
Procell is expensive and toxicity is bigger.
Chemiluminescent buffer Procell is tripropyl amine (TPA) cleanout fluid, is used for safeguarding electrode, transporting reactant mixture, cleaning
The coated microgranule of Streptavidin, generation reaction signal；Chemiluminescence cleanout fluid Cleancell, pipeline after every time measuring
System and the cleaning of measuring cell and electrodes maintenance.Roche Diagnistics's import registration certificate information display Procell formula is: phosphate
Buffer 300mmol/L, tripropyl amine (TPA) 180mmol/L, detergent 0.1%；Preservative, pH6.8；Cleancell formula is:
KOH 176mmol/L (is equivalent to pH value 13.2)；Cleaning agent≤1%.Tris (bipyridine) ruthenium ([Ru (bpy) 32+])-tripropyl amine (TPA)
(TPrA) system is a kind of common ECL system, and wherein tris (bipyridine) ruthenium is electrochemiluminescence agent, and it and electron donor TPrA exist
Electrode anode surface loses electronics generation oxidation reaction simultaneously, [the Ru (bpy) 3 of divalent2+] label is oxidized to the [Ru of trivalent
(bpy)33+] label, TPrA is oxidized to radical cation TPrA+*, TPrA+*The most unstable, spontaneously lose one
Proton and form free radical TPrA*, it is strong reductant, by an electronics to the [Ru (bpy) 3 of trivalent3+] so that it is become and excite
The Ru (bpy) 3 of state2+, and TPrA self is oxidized to oxidation product.The Ru (bpy) 3 of excited state2+A wavelength is launched during decay
The photon of 620nm, re-forms the Ru (bpy) 3 of ground state2+.The carrying out that this process is gone round and begun again at electrode surface, produces many
Photon so that optical signal strengthens.
Due to TprA autoxidation irreversibility and between the two interact produce light more weak, this system lead to
Often use large excess of TPrA (typically TPrA:Ru (bpy) 32+=100000 μm ol/L:1 μm ol/L), i.e. 100mmol/L,
And original-pack reagent adds 180mmol/L TPrA, beyond usual amounts, and TPrA toxicity is designated high toxicity, rat oral LD50
=72mg/kg, its steam or mist have strong impulse to eye, mucosa, upper respiratory tract and skin, for the healthy shadow of docimaster
Ring bigger.Additionally, full automatic biochemical apparatus is after a number of uses, owing to cleaning performance is the best, from measuring cell to pipeline, it is easily subject to
The pollution of reactant liquor, long-term protein precipitation, cause blank absorbency to increase, and occur reporting to the police.
Visible, further research can be used for chemiluminescent buffer and the cleanout fluid of Roche automatic lmunoassays analyzer,
Especially reduce the concentration of tripropyl amine (TPA), and ensure the concordance of testing result, stability and the buffer of accuracy, and have
The cleanout fluid of more excellent cleaning performance, remain technical problem urgently to be resolved hurrily, this most just the present invention be accomplished
Place, basis and power are leaned on.
Summary of the invention
As it has been described above, in order to further study the chemiluminescent buffer that can be used for Roche automatic lmunoassays analyzer and
Cleanout fluid, especially reduces the concentration of tripropyl amine (TPA), and ensures the concordance of testing result, stability and the buffer of accuracy,
And there is the cleanout fluid of more excellent cleaning performance, this is conducted in-depth research by the present inventor, is paying a large amount of wound
After the property made work, thus complete the present invention.
The present invention is with regard to TPrA Yu Ru (bpy) 32+The light produced that interacts more weak set about, Procell formula is changed
Enter, be substantially reduced TPrA consumption, in prescription, add N, N-bis-[just] butylamino ethanol, sodium chloride, PEG 300, third simultaneously
The material such as diureide, paracresol, enhanced chemiluminescence, reduce and weaken because tripropyl amine (TPA) concentration reduces the chemiluminescence caused, meanwhile,
Ensure the concordance with original-pack reagent and stability.
The present invention adds the composition such as carbamide, thiourea Trixton X-100, MES in Cleancell, increases it to condensation
The cleaning performance of the composition such as block and fibrin, and add Proclin 300 preservative, safeguard electrode further.
Procell and Cleancell developing improvement formula substitutes original-pack Procell and Cleancell, to reduce poison
Property, improve cleaning performance, further shield electrode, reduce cost, there is bigger profit margin.
Specifically, the present invention relates to following four aspects, more specifically, first aspect, the present invention relates to one
Matching used electrochemiluminescence buffer and cleanout fluid, buffer composition includes: sodium dihydrogen phosphate, tripropyl amine (TPA), sodium chloride, grass
Acid, Tween 20, Proclin 300；Cleanout fluid component includes: potassium hydroxide, Trixton X-100, MES, Proclin
300。
Preferably, its component of buffer therein and content is: potassium dihydrogen phosphate 250～350mmol/L, tripropyl amine (TPA) 60～
90mmol/L, sodium chloride 1～10mmol/L, oxalic acid 3～10mmol/L, Tween 20 0.05～0.2% (w%), Proclin
300 0.05～0.2% (w%), pH value 6.6～7.0；Cleanout fluid component therein and content is: potassium hydroxide 150-
200mmol/L, Trixton X-100 0.1～1% (w%), MES 0.1～1% (w%), Proclin 300 0.1～
0.5% (w%).
Second aspect, the present invention relates to a kind of matching used electrochemiluminescence buffer and cleanout fluid, buffer group
Divide and include: sodium dihydrogen phosphate, tripropyl amine (TPA), sodium chloride, PEG 300, Tween 20, Proclin 300, paracresol；Cleanout fluid group
Divide and include: potassium hydroxide, Trixton X-100, MES, carbamide, Proclin 300.
Preferably, its component of buffer therein and content is: potassium dihydrogen phosphate 250～350mmol/L, tripropyl amine (TPA) 20～
30mmol/L, sodium chloride 1～10mmol/L, PEG300 1～10mmol/L, Tween 20 0.05～0.2% (w%),
Proclin 3000.05～0.2% (w%), paracresol 0.1～1mmol/L, pH value 6.6～7.0；Cleanout fluid component therein
With content it is: potassium hydroxide 150-200mmol/L, Trixton X-100 0.1～1% (w%), MES/AES 0.1～1%
(w%), carbamide 0.1～1% (w%), Proclin300 0.1～0.5% (w%).
3rd aspect, the present invention relates to a kind of matching used electrochemiluminescence buffer and cleanout fluid, buffer group
Divide and include: sodium dihydrogen phosphate, tripropyl amine (TPA), N, N-bis-[just] butylamino ethanol, sodium chloride, PEG 300, malonyl urea, Tween
20, Proclin 300, paracresol, polyvinylpyrrolidone, methylcellulose；Cleanout fluid component includes: potassium hydroxide,
Trixton X-100, MES, thiourea, Proclin 300.
Preferably, its component of buffer therein and content is: potassium dihydrogen phosphate 250～350mmol/L, tripropyl amine (TPA) 1～
10mmol/L, N, N-bis-[just] butylamino ethanol 1～3mmol/L, sodium chloride 1～10mmol/L, PEG 300 1～
10mmol/L, malonyl urea 1～5mmol/L, Tween 20 0.05～0.2% (w%), Proclin 300 0.05～0.2%
(w%), paracresol 0.1～1mmol/L, polyvinylpyrrolidone 0.1～1mmol/L, methylcellulose 0.1～1mmol/L, pH
Value 6.6～7.0；Cleanout fluid component therein and content is: potassium hydroxide 150-200mmol/L, Trixton X-100 0.1～
1% (w%), MES0.1～1% (w%), thiourea 0.1～1% (w%), Proclin 300 0.1～0.5% (w%).
4th aspect, the present invention relates to the compound method of above-mentioned buffer and cleanout fluid, as follows:
One, the preparation of chemiluminescent buffer: weigh respectively to add in a small amount of water by the composition in addition to tripropyl amine (TPA) and stir
Mix and make dissolving, add tripropyl amine (TPA) stirring standing and be transferred to after the most molten in volumetric flask, constant volume, shake up, regulate pH value, to obtain final product.
Two, the preparation of cleanout fluid: weigh suitable amount according to the composition in formula, be dissolved in water mixing, lets cool, is transferred to
Volumetric flask, constant volume, shake up, to obtain final product.
Detailed description of the invention
Below by specific embodiment, the present invention is described in detail, but the purposes of these exemplary embodiments and
Purpose is only used for enumerating the present invention, and not the real protection scope to the present invention constitutes any type of any restriction, more non-general
Protection scope of the present invention is confined to this.
Embodiment 1
A, the component of chemiluminescent buffer (Procell) and preparation:
Potassium dihydrogen phosphate 270mmol/L
Tripropyl amine (TPA) 60mmol/L
Sodium chloride 2mmol/L
Oxalic acid 5mmol/L
Tween 20 0.05% (w%)
Proclin 300 0.05% (w%)
PH value NaOH is adjusted to 6.6
Weigh respectively to add in a small amount of water by other compositions in addition to tripropyl amine (TPA) and be stirred to dissolve, add tripropyl amine (TPA) and stir
Mix standing to move to after the most molten in volumetric flask, constant volume, shake up and get final product.
B, the component of cleanout fluid (Cleancell) and preparation:
Potassium hydroxide 150mmol/L
Trixton X-100 0.5%
MES 0.5%
Proclin 300 0.3%
Weighing suitable amount according to the composition in formula, be dissolved in water mixing, lets cool, and is transferred to volumetric flask, constant volume, shakes up
Obtain.
Embodiment 2
A, the component of buffer (Procell) and preparation:
Potassium dihydrogen phosphate 350mmol/L
Tripropyl amine (TPA) 25mmol/L
Sodium chloride 5mmol/L
PEG 300 5mmol/L
Tween 20 0.1% (w%)
Proclin 300 0.1% (w%)
Paracresol 0.2mmol/L
PH value NaOH is adjusted to 7.0
Weigh respectively to add in a small amount of water by other compositions in addition to tripropyl amine (TPA) and be stirred to dissolve, add tripropyl amine (TPA) and stir
Mix standing to move to after the most molten in volumetric flask, constant volume, shake up and get final product.
B, the component of cleanout fluid (Cleancell) and preparation:
Potassium hydroxide 200mmol/L
Trixton X-100 0.2%
AES 0.8%
Carbamide 0.1%
Proclin 300 0.5%
Weighing suitable amount according to the composition in formula, be dissolved in water mixing, lets cool, and is transferred to volumetric flask, constant volume, shakes up
Obtain.
Embodiment 3
A, the component of buffer (Procell) and preparation:
Potassium dihydrogen phosphate 300mmol/L
Tripropyl amine (TPA) 5mmol/L
N, N-bis-[just] butylamino ethanol 3mmol/L
Sodium chloride 5mmol/L
PEG 300 3mmol/L
Malonyl urea 5mmol/L
Tween 20 0.1% (w%)
Proclin 300 0.05% (w%)
Paracresol 0.1mmol/L
Polyvinylpyrrolidone 0.1mmol/L
Methylcellulose 0.1mmol/L
PH value NaOH is adjusted to 6.8
Weigh respectively to add in a small amount of water by other compositions in addition to tripropyl amine (TPA) and be stirred to dissolve, add tripropyl amine (TPA) and stir
Mix standing to move to after the most molten in volumetric flask, constant volume, shake up and get final product.
B, the component of cleanout fluid (Cleancell) and preparation:
Potassium hydroxide 180mmol/L
Trixton X-100 0.8%
MES 0.2%
Thiourea 0.5%
Proclin 300 0.5%
Weighing suitable amount according to the composition in formula, be dissolved in water mixing, lets cool, and is transferred to volumetric flask, constant volume, shakes up
Obtain.
Embodiment 4
One, Accuracy Verification test is measured
1, Procell and Cleancell that embodiment 1 is configured is placed in the Elecsys2010 electrochemiluminescence of Roche
Carry out checking test on the corresponding position of immunoassay system, take 30 patients serums and test, contrast and original-pack supporting examination
The dependency of agent Procell, Cleancell testing result.
Result shows, Procell, Cleancell of autogamy embodiment 1 and original-pack Procell, Cleancell performance one
Cause is good.
2, Procell and Cleancell that embodiment 2 is configured is placed in the Elecsys2010 electrochemiluminescence of Roche
Carry out checking test on the corresponding position of immunoassay system, take 30 patients serums and test, contrast and original-pack supporting examination
The dependency of agent Procell, Cleancell testing result.
Result shows, autogamy embodiment 2Procell, Cleancell are consistent with original-pack Procell, Cleancell performance
Property is good.
3, Procell and Cleancell that embodiment 3 is configured is placed in the Elecsys2010 electrochemiluminescence of Roche
Carry out checking test on the corresponding position of immunoassay system, take 30 patients serums and test, contrast and original-pack supporting examination
The dependency of agent Procell, Cleancell testing result.
Result shows, autogamy embodiment 3Procell, Cleancell are consistent with original-pack Procell, Cleancell performance
Property is good.
In summary it can be seen, show that Self-made reagent, in the case of greatly reducing tripropyl amine (TPA) concentration, has remained to simultaneously
Immunoassay on fully-automatic analyzer, meets and the conforming goldstandard of original-pack reagent, and result is accurately and reliably, it is seen that this
Procell, Cleancell of improving formula of invention have quite excellent safety and reliability.
Embodiment 5
Two, Cleancell cleaning performance checking test
The effect of Cleancell mainly every time measure after tubing and the cleaning of measuring cell and electrodes maintenance, this
Invention on the basis of the formula of original-pack reagent, improve formula, with improve its to pipeline and the cleaning action of measuring cell, to therein
The albumen of reactant liquor mainly residual is carried out, and is first reacted with thyrotropin test kit by a large amount of blank serum, then will
Above reactant liquor is sub-packed in blank cuvette, stands 30min, outwells, and uses autogamy Cleancell and imported reagents to exist
Carry out cuvette cleaning (respectively carrying out 20 groups of parallel tests) in the case of identical addition, add the water of equivalent after cleaning, shake
Even, under protein maximum absorption wavelength 280nm, test absorbance, observed result.
1. being tested according to above verification method by the Cleancell configured in embodiment 1, result see table:
2. being tested according to above verification method by the Cleancell configured in embodiment 2, result see table:
3. being tested according to above verification method by the Cleancell configured in embodiment 3, result see table:
In summary it can be seen, autogamy embodiment 1-3, improve the Cleancell of formula compared with imported reagents
The albumen cleaning performance of Cleancell is more excellent, therefore the Cleancell improving formula of the present invention, and cleaning action strengthens.Thus
Making, the present invention matching used electrochemiluminescence buffer and cleanout fluid will have more the industrial application value of reality.
Should be appreciated that the purposes of these embodiments is merely to illustrate the present invention and is not intended to limit the protection model of the present invention
Enclose.Additionally, it will also be appreciated that after the technology contents having read the present invention, the present invention can be made respectively by those skilled in the art
Planting change, amendment and/or modification, all these equivalent form of value falls within the guarantor that the application appended claims is limited equally
Within the scope of protecting.
Claims (6)
1. a matching used electrochemiluminescence buffer and cleanout fluid, it is characterised in that: buffer composition includes: di(2-ethylhexyl)phosphate
Hydrogen sodium, tripropyl amine (TPA), sodium chloride, oxalic acid, Tween 20, Proclin 300；Cleanout fluid component includes: potassium hydroxide, Trixton
X-100, MES, Proclin 300.
2. buffer as claimed in claim 1 and cleanout fluid, it is characterised in that its component of buffer therein and content is:
Potassium dihydrogen phosphate 250～350mmol/L, tripropyl amine (TPA) 60～90mmol/L, sodium chloride 1～10mmol/L, oxalic acid 3～10mmol/
L, Tween20 0.05～0.2% (w%), Proclin 300 0.05～0.2% (w%), pH value 6.6～7.0；Therein clearly
Washing liquid component and content is: potassium hydroxide 150-200mmol/L, Trixton X-100 0.1～1% (w%), MES 0.1～
1% (w%), Proclin 3000.1～0.5% (w%).
3. a matching used electrochemiluminescence buffer and cleanout fluid, it is characterised in that: buffer composition includes: di(2-ethylhexyl)phosphate
Hydrogen sodium, tripropyl amine (TPA), sodium chloride, PEG 300, Tween 20, Proclin 300, paracresol；Cleanout fluid component includes: hydroxide
Potassium, Trixton X-100, MES, carbamide, Proclin 300.
4. buffer as claimed in claim 3 and cleanout fluid, it is characterised in that its component of buffer therein and content is:
Potassium dihydrogen phosphate 250～350mmol/L, tripropyl amine (TPA) 20～30mmol/L, sodium chloride 1～10mmol/L, PEG 300 1～
10mmol/L, Tween 20 0.05～0.2% (w%), Proclin 300 0.05～0.2% (w%), paracresol 0.1～
1mmol/L, pH value 6.6～7.0；Cleanout fluid component therein and content is: potassium hydroxide 150-200mmol/L, Trixton
X-100 0.1～1% (w%), MES 0.1～1% (w%), carbamide 0.1～1% (w%), Proclin 300 0.1～
0.5% (w%).
5. a matching used electrochemiluminescence buffer and cleanout fluid, it is characterised in that: buffer composition includes: di(2-ethylhexyl)phosphate
Hydrogen sodium, tripropyl amine (TPA), N, N-bis-[just] butylamino ethanol, sodium chloride, PEG 300, malonyl urea, Tween 20, Proclin
300, paracresol, polyvinylpyrrolidone, methylcellulose；Cleanout fluid component includes: potassium hydroxide, Trixton X-100,
MES, thiourea, Proclin 300.
6. buffer as claimed in claim 5 and cleanout fluid, it is characterised in that its component of chemiluminescent buffer therein and
Content is: potassium dihydrogen phosphate 250～350mmol/L, tripropyl amine (TPA) 1～10mmol/L, N, N-bis-[just] butylamino ethanol 1～
3mmol/L, sodium chloride 1～10mmol/L, PEG 300 1～10mmol/L, malonyl urea 1～5mmol/L, Tween 20
0.05～0.2% (w%), Proclin 300 0.05～0.2% (w%), paracresol 0.1～1mmol/L, polyvinylpyrrolidine
Ketone 0.1～1mmol/L, methylcellulose 0.1～1mmol/L, pH value 6.6～7.0；Cleanout fluid component therein and content is:
Potassium hydroxide 150-200mmol/L, Trixton X-100 0.1～1% (w%), MES 0.1～1% (w%), thiourea 0.1～
1% (w%), Proclin 300 0.1～0.5% (w%).
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| CN108303528A (en) * | 2018-01-19 | 2018-07-20 | 青岛大学附属医院 | Substrate and its preparation method and application used in a kind of electrochemiluminescence analysis instrument |
| CN109517672A (en) * | 2017-09-20 | 2019-03-26 | 刘宏飞 | A kind of Roche Electrogenerated chemiluminescent immunoassay instrument cleaning solution and preparation method thereof |
| CN111537706A (en) * | 2020-05-26 | 2020-08-14 | 长春晨裕生物医疗科技有限公司 | Immune lotion for electrochemical luminescence immunoassay instrument |
| CN111896731A (en) * | 2020-06-24 | 2020-11-06 | 宁波瑞源生物科技有限公司 | Cleaning solution for chemiluminescence immunoassay |
| CN112924438A (en) * | 2021-01-26 | 2021-06-08 | 金华市鑫科医疗器械有限公司 | Chemiluminescent cleaning buffer solution |
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| CN108303528A (en) * | 2018-01-19 | 2018-07-20 | 青岛大学附属医院 | Substrate and its preparation method and application used in a kind of electrochemiluminescence analysis instrument |
| US11927592B2 (en) | 2019-01-03 | 2024-03-12 | Meso Scale Technologies, Llc. | Compositions and methods for carrying out assay measurements |
| CN111537706A (en) * | 2020-05-26 | 2020-08-14 | 长春晨裕生物医疗科技有限公司 | Immune lotion for electrochemical luminescence immunoassay instrument |
| CN111896731A (en) * | 2020-06-24 | 2020-11-06 | 宁波瑞源生物科技有限公司 | Cleaning solution for chemiluminescence immunoassay |
| CN112924438A (en) * | 2021-01-26 | 2021-06-08 | 金华市鑫科医疗器械有限公司 | Chemiluminescent cleaning buffer solution |
| CN116286205A (en) * | 2022-12-23 | 2023-06-23 | 浙江鑫科医疗科技有限公司 | Tripropylamine cleaning buffer solution matched with Rogowski Cobas e801 chemiluminescent immunoassay analyzer and preparation method thereof |
| CN116286205B (en) * | 2022-12-23 | 2024-04-19 | 浙江鑫科医疗科技有限公司 | Tripropylamine buffer solution matched with Luo Cobas e801 chemiluminescent immunoassay analyzer and preparation method thereof |
| CN118275417A (en) * | 2024-03-20 | 2024-07-02 | 江苏昱安生物科技有限公司 | Buffer solution for Roche electrochemiluminescence immunoassay instrument and preparation method thereof |
| CN118275417B (en) * | 2024-03-20 | 2024-09-17 | 江苏昱安生物科技有限公司 | Buffer solution for Roche electrochemiluminescence immunoassay instrument and preparation method thereof |
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