CN108303528A - Substrate and its preparation method and application used in a kind of electrochemiluminescence analysis instrument - Google Patents
Substrate and its preparation method and application used in a kind of electrochemiluminescence analysis instrument Download PDFInfo
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- CN108303528A CN108303528A CN201810054857.XA CN201810054857A CN108303528A CN 108303528 A CN108303528 A CN 108303528A CN 201810054857 A CN201810054857 A CN 201810054857A CN 108303528 A CN108303528 A CN 108303528A
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- polymerization
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- substrate
- polyoxy vinethene
- ethoxylated dodecyl
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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
- G01N21/763—Bioluminescence
Abstract
The invention discloses substrates and its preparation method and application used in a kind of electrochemiluminescence analysis instrument, it is related to electrochemical analysis field, including luminescence buffer, measuring cell cleaning solution and prerinse liquid, luminescence buffer includes that the degree of polymerization is the isooctanol polyoxy vinethene of (12 16), the ethoxylated dodecyl alcohol that the degree of polymerization is (6 10).Inventive substrate measurement range is identical as original product, as a result accurately and reliably, prepares simply, of low cost, stability is long.
Description
Technical field
The present invention relates to electrochemical analysis fields, and in particular to substrate and its preparation used in a kind of electrochemiluminescence analysis instrument
Methods and applications.
Background technology
Electrogenerated chemiluminescent immunoassay technology is one kind in the immuno analytical method of Roche Holding Ag of Switzerland invention.It is extensively
Using the quantitative determination of Thyreoidine, sex hormone, bone metabolism, myocardial infarction, tumor markers, infectious disease antigen antibody etc..Its
Based on reaction principle be antigen-antibody immune response.
Reaction process is as follows：1. instrument draws the biotin reagent containing antibody, antigen or enzyme and is coated with strepto-
The magnetic microsphere compound of Avidin, is added to reaction cup mixing, is incubated at 37 DEG C and reacts 9~27min in disk, makes test substance
It is combined with antibody, antigen or the enzyme spcificity after biotinylation；2. being used in conjunction with after the completion ofPrerinse liquid(abbreviation PreClean M)
Potential interfering substance is removed, unbonded ingredient is made to be removed before compound is admitted to measuring cell；3. probe is inhaled again
It takes containing tripropyl amine (TPA)Luminescence bufferInto measuring cell, a concentration of 180mmol/L of tripropyl amine (TPA) wherein in luminescence buffer；
4. finally cleaning entire pipeline with measuring cell cleaning solution (CleanCell).
Reaction mechanism is：For determinand under the action of electric field, luminous marker loses an electronics oxidation in reaction solution
At the tris (bipyridine) ruthenium of trivalent, while tripropyl amine (TPA) also loses an electronics and is aoxidized, and is dehydrogenated to tripropyl amine (TPA) free radical, tripropyl amine (TPA) is certainly
An electronics is transmitted by base to be allowed to become excitation state to trivalent tris (bipyridine) ruthenium；And the second bipyridine ruthenium in excitation state is unstable
Fixed, the formation with one wavelength of transmitting for 620nm photons, which releases energy, returns to ground state, and tris (bipyridine) ruthenium is not consumed, then is connect
Electronic circulation by the transmission of tripropyl amine (TPA) shines.Photomultiplier mounted on measuring cell top receives the light letter of 620nm photons
Number, and electric signal is converted thereof into, the amplitude relationship proportional to testing concentration or content of electric signal, finally by calculating
Go out the concentration or content of measured substance.
In electrochemical luminescence whole process, need complete using luminescence buffer, detection cell cleaning solution, prerinse liquid ability
At measurement, and the original-pack substrate of Roche is of high cost.Therefore it is allowed to domesticize and reach that result is accurate and reliable, reduces testing cost tool
There is important Social benefit and economic benefit.
It is learnt through retrieval, on 2 18th, 2015 Publication No. CN104360051A, Publication No. on June 10th, 2015
The patent application of CN104698164A, the November in 2016 of Publication No. CN106124753A on the 16th have carried out correlative study, but its
Technically there is problem and risk.Because during electrochemical luminescence, tripropyl amine (TPA) is oxidized to di-n-propylamine and propionic aldehyde, itself
It is constantly consumed, the concentration of test substance is higher, needs the amount of the tripropyl amine (TPA) consumed more.The original-pack of Roche Holding Ag shines
The tripropyl amine (TPA) of buffer solution (ProCell) containing concentration 180mmol/L, just allows in the human sample of clinical examination often
The test substance for high concentration occur, needs the up to tripropyl amine (TPA) of 180mmol/L that can just meet clinical needs in Electrochemical Detection,
If such concentration is not achieved in the tripropyl amine (TPA) concentration in luminescence buffer, measurement range reduction is directly resulted in, immune response occurs
Hook effect, the patient's sample result of high concentration will be inaccurate, causes Lower result, or even false negative result occur, makes
Risk is failed to pinpoint a disease in diagnosis at clinic, is delayed the diagnosing and treating of patient.
Wherein, the tripropyl amine (TPA) for being 2.6% containing volumetric concentration in the formula of Publication No. CN104360051A, mass concentration
For 19.63g/L, the 76% of a concentration of original-pack concentration of Roche, tripropyl amine (TPA) 12.0- in the formula of Publication No. CN104698164A
16.0g/L, the 46~62% of a concentration of original-pack concentration, tripropyl amine (TPA) a concentration of 60 in the formula of Publication No. CN106124753A~
90mmol/L, the 33%~50% of a concentration of original-pack concentration.In these formulations, it is original-pack to be all not up to Roche for the concentration of tripropyl amine (TPA)
180mmol/L, cause the clinical sample testing result of high concentration inaccurate, bring and potentially fail to pinpoint a disease in diagnosis risk.
Invention content
Existing luminescence buffer discloses tripropyl amine (TPA) concentration in formula and causes less than original-pack concentration for the above-mentioned prior art
Measurement range reduces, the problem of high concentration clinical sample testing result inaccuracy, and the purpose of the present invention is to provide a kind of electrifications
Substrate and its preparation method and application used in luminescence analyzer is learned, inventive substrate measurement range is identical as original product, as a result
Accurately and reliably, it prepares simply, of low cost, stability is long.
Substrate used in a kind of electrochemiluminescence analysis instrument, including luminescence buffer, measuring cell cleaning solution and prerinse liquid, hair
Light buffer solution includes that the degree of polymerization is the isooctanol polyoxy vinethene of (12-16), the laruyl alcohol polyoxy second that the degree of polymerization is (6-10)
Alkene ether.
Specifically, luminescence buffer include the degree of polymerization be the isooctanol polyoxy vinethene of (12-16), the degree of polymerization is (6-
10) concentrated phosphoric acid of ethoxylated dodecyl alcohol, 85wt%, 2 centinormal 1 dilute hydrochloric acid, dichloro acetamide, potassium dihydrogen phosphate
And tripropyl amine (TPA).
Luminescence buffer is further preferably：
Phosphoric acid (2-5) g/L of 85wt%,
2 centinormal 1 hydrochloric acid (0.1-1.0) g/L,
Dichloro acetamide 1-3g/L,
The degree of polymerization is isooctanol polyoxy vinethene (0.6-1.2) g/L of (12-16),
The degree of polymerization is ethoxylated dodecyl alcohol (0.8-1.5) g/L of (6-10),
Potassium dihydrogen phosphate (35-40) g/L, and
Tripropyl amine (TPA) (20-30) g/L.
Specifically, measuring cell cleaning solution includes that the degree of polymerization is for the isooctanol polyoxy vinethene of (12-16), the degree of polymerization
The ethoxylated dodecyl alcohol and 85wt% potassium hydroxide aqueous solutions of (6-10).
Measuring cell cleaning solution is further preferably：
The degree of polymerization is isooctanol polyoxy vinethene (0.8-1.2) g/L of (12-16),
The degree of polymerization is ethoxylated dodecyl alcohol (8-12) g/L of (6-10), and
Potassium hydroxide aqueous solution (8-12) g/L of 85wt%.
Specifically, prerinse liquid is phosphate buffer, and phosphoric acid, dichloro acetamide containing 85wt%, the degree of polymerization are
The isooctanol polyoxy vinethene and the degree of polymerization of (12-16) are the ethoxylated dodecyl alcohol of (6-10).
Prerinse liquid is further preferably
Sodium dihydrogen phosphate dihydrate (0.5-1.0) g/L,
Disodium hydrogen phosphate (2.0-2.5) g/L,
Sodium chloride (1.0-1.5) g/L,
Dichloro acetamide (1.0-2.0) g/L,
85wt% phosphoric acid (180-220) mg/L,
The degree of polymerization is isooctanol polyoxy vinethene (0.8-1.2) g/L of (12-16), and
The degree of polymerization is ethoxylated dodecyl alcohol (0.8-1.2) g/L of (6-10).
The present invention also provides a kind of preparation methods of substrate used in above-mentioned electrochemiluminescence analysis instrument, include the following steps：
1. preparing luminescence buffer：Each required component in addition to tripropyl amine (TPA) is weighed according to formula, is mixed with 700 milliliters of water, stirring and dissolving；Add
Enter tripropyl amine (TPA), stirs 10 minutes, be settled to 1000 milliliters, mixing；2. preparing measuring cell cleaning solution：It is weighed according to formula each required
Component, mixes with 700 milliliters of water, and stirring and dissolving is settled to 1000 milliliters, mixing；3. preparing prerinse liquid：It is weighed according to formula
Component needed for each, mixes, stirring and dissolving is settled to 1000 milliliters, mixing with 700 milliliters of water.
Preferably, the isooctanol polyoxy vinethene degree of polymerization is 14, and the ethoxylated dodecyl alcohol degree of polymerization is 9.
A kind of application of low polymerization degree surfactant in preparing substrate used in electrochemiluminescence analysis instrument, the oligomerization
Degree surfactant includes that the laruyl alcohol that the degree of polymerization is the isooctanol polyoxy vinethene of (12-16), the degree of polymerization is (6-10) is poly-
Ethylene oxide ether.
Preferably, the isooctanol polyoxy vinethene degree of polymerization is 14, and the ethoxylated dodecyl alcohol degree of polymerization is 9.
The present invention uses the low polymerization degree surfactant of small-molecular-weight, preferably selects the oligomeric state laruyl alcohol being in a liquid state poly-
Ethylene oxide ether and isooctanol polyoxy vinethene improve micelle state, control emulsifying power, and avoid gelation.Especially increase
Tripropyl amine (TPA) rate of dissolution in luminescence buffer process for preparation makes the dynamic concentration of tripropyl amine (TPA) be not less than the original-pack luminous buffering of Roche
Liquid, and matched with phosphoric acid and hydrochloric acid, increase the sensitivity of electrochemical luminescence, joint ensures measurement range, while using dichloro
Acetamide ensures that the stability of luminescence buffer, the term of validity can reach 24 months as preservative.
Two kinds of surfactants contained by measuring cell cleaning solution of the present invention are in liquid condition, soluble, have excellent emulsification
And wetting power, reduction surface tension is with obvious effects, can completely remove the pollutant in reaction system, ensures to measure the clear of pipeline
Wash effect.It need not heat and cool down in process for preparation simultaneously, and preservative need not be added, keep process for preparation simple, easily
In production.
Prerinse liquid of the present invention not only makes the acid-base value of buffer system maintain neutrality, and can thoroughly remove potential
Interfering substance makes unbonded ingredient be removed before immune complex is admitted to measuring cell, ensures the specificity of result
And accuracy.
Resultant effect caused by the present invention includes：
The complete substrate of the excellent luminescence buffer of inventive formulation, measuring cell cleaning solution and prerinse liquid composition, can
Ensure clinical sample measurement range and original-pack consistent, raising tripropyl amine (TPA) rate of dissolution, detection accuracy height, formula and preparation method
Simple and practicable, performance is stablized, effective up to 24 months, saves Expenses of laboratory examination 60%, has significant clinical meaning and economic effect
Benefit.
Description of the drawings
Fig. 1 is that the THS measurement ranges of substrate verification test of the embodiment of the present invention verify regression figure.
Fig. 2 is that the E2 measurement ranges of substrate verification test of the embodiment of the present invention verify regression figure.
Fig. 3 is that the PROG measurement ranges of substrate verification test of the embodiment of the present invention verify regression figure.
Fig. 4 is that the AFP measurement ranges of substrate verification test of the embodiment of the present invention verify regression figure.
Fig. 5 is that the THS measurement ranges of substrate verification test of the embodiment of the present invention verify regression figure.
Fig. 6 is the THS Accuracy Verification regression figures of substrate verification test of the embodiment of the present invention.
Fig. 7 is the E2 Accuracy Verification regression figures of substrate verification test of the embodiment of the present invention.
Fig. 8 is the PROG Accuracy Verification regression figures of substrate verification test of the embodiment of the present invention.
Fig. 9 is the AFP Accuracy Verification regression figures of substrate verification test of the embodiment of the present invention.
Figure 10 is the THS Accuracy Verification regression figures of substrate verification test of the embodiment of the present invention.
Specific implementation mode
The specific implementation mode that the following embodiment of the present invention only is used for illustrating to realize the present invention, these embodiments cannot
It is not understood as limitation of the present invention.It is other it is any the change made without departing from the spirit and principles of the present invention,
Modification substitutes, combination, simplifies, and is accordingly to be regarded as equivalent substitute mode, falls within the scope and spirit of the invention.
Embodiment
Substrate and preparation method thereof used in a kind of electrochemiluminescence analysis instrument should include that luminescence buffer, measuring cell clean
Liquid and prerinse liquid.
1. luminescence buffer (abbreviation Procell) forms and preparation method is：
Composition：The phosphoric acid 3.2g/L of 85wt%,
2 centinormal 1 hydrochloric acid 0.5g/L,
Dichloro acetamide 1.5g/L,
The isooctanol polyoxy vinethene 1.0g/L that the degree of polymerization is 14,
The ethoxylated dodecyl alcohol 1.2g/L that the degree of polymerization is 9,
Potassium dihydrogen phosphate 39g/L, and
Tripropyl amine (TPA) 26g/L.
It prepares：Beaker is placed on balance, zero setting, is then weighed according to above-mentioned formula composition required in addition to tripropyl amine (TPA)
Component is added in beaker, and 700 milliliters of water are added, and is stirred 5 minutes and is dissolved, tripropyl amine (TPA) is added, stirs 10 minutes, be settled to
It is 1000 milliliters, existing after mixing.
2. measuring cell cleaning solution (abbreviation CleanCell) forms and preparation method is：
Composition：The isooctanol polyoxy vinethene 1.0g/L that the degree of polymerization is 14,
The ethoxylated dodecyl alcohol 10g/L that the degree of polymerization is 9, and
The potassium hydroxide aqueous solution 10.5g/L of 85wt%.
It prepares：Beaker is placed on balance, zero setting, then weighing required component according to formula is added in beaker,
700 milliliters of water are added, stirs 5 minutes and dissolves, be settled to 1000 milliliters, it is existing after mixing.
3. prerinse liquid (abbreviation PreClean M) is phosphate buffer, composition and preparation method are：
Composition：Sodium dihydrogen phosphate dihydrate 0.7g/L,
Disodium hydrogen phosphate 2.2g/L,
Sodium chloride 1.35g/L,
Dichloro acetamide 1.5g/L,
85wt% phosphoric acid 200mg/L,
The isooctanol polyoxy vinethene 1.0g/L that the degree of polymerization is 14, and
The ethoxylated dodecyl alcohol 1.0g/L that the degree of polymerization is 9.
It prepares：Beaker is placed on balance, zero setting, then weighing required component according to formula is added in beaker,
700 milliliters of water are added, stirs 5 minutes and dissolves, be settled to 1000 milliliters, it is existing after mixing.
4. by the above-mentioned substrate of preparation, including luminescence buffer, measuring cell cleaning solution and prerinse liquid are positioned over Roche electricity
It is tested and analyzed for electrochemical luminescence on chemical e601 analyzers.
5. placing analytical reagent and quality-control product and clinical sample, start 601 analyzer of electrochemical luminescence, without school again
Standard is directly detected.
6. recording result after the completion of detection.
Verification test
By above-described embodiment substrate for electrochemiluminescence analysis separately verify its precision, measurement range, accuracy and
Stability.
Verification Project：The present invention verifies precision and selects with the representative project of apparent clinical meaning.
Thyroid hormone, abbreviation TSH is a kind of protein that molecular weight is 30kD, and TSH is thermophilic in prehypophyseal specificity
Alkali generates into the cell.TSH detections are the Primary Screening Tests for finding out thyroid function.The minor change of free thyroid concentration will band
Carry out notable adjustment of the TSH concentration to negative direction, TSH is the very sensitive specificity parameter for testing thyroid function, especially suitable
Together in the dysfunction of early detection or exclusion Hypothalamus-Pituitary-Thyroid central regulation loop.
Estradiol, abbreviation E2 are strongest estrogen, are mainly generated by ovary.Detection estradiol can be used for explaining inferior colliculus
Brain-pituitary-sexual gland regulatory function disorder, generates on the ovary and orchioncus and kidney of estrogenic gynecomastia
Gland corticohyperplassia etc..
Progesterone, abbreviation PROG belong to steroid hormone, molecular weight 314.5D, mainly in the cell of corpus luteum and the gestational period
Placenta in formed, the concentration of progesterone and the growth of corpus luteum and degenerate closely related.Progesterone can make uterine mucosa be transformed into gland
The abundant tissue of body (secretory phase).Progesterone is measured to be diagnosed for reproduction：The detection of the onset of ovulation and the estimation of luteal phase.
Alpha-fetoprotein, abbreviation AFP derive from yolk bag, undifferentiated liver cell and Gastrointestinal Tract of Fetus.70-95%'s is primary
Property liver cancer patient AFP increase, late period, AFP contents are higher.
Prostate-specific antigen, abbreviation tPSA belong to glycoprotein, and increasing general prompt prostate, there are lesions, such as forefront
Adenositis, hyperplasia of prostate or cancer.
One, precision is verified：
1 verification meaning：Good precision is the premise of measurement range and accuracy, thus firstly the need of verify its
Precision.
2 verification methods：The present embodiment is according to National Committee for Clinical Laboratory Standards (abbreviation NCCLS) EP5- in the U.S.
The evaluation of A-- clinical chemistry equipment operation precision carries out precision verification.
2.1 quality-control product sources：It is raw that normal value quality-control product and exceptional value quality-control product are all made of Bio Rad Laboratories (Bio-RAD)
The mating quality controlled serum of the electrochemical luminescence e601 analyzers of production.
2.2 withinrun precisions are tested：It chooses normal value quality-control product and exceptional value quality-control product is continuously done 20 times, calculating mean value,
Standard deviation, the coefficient of variation (CV%).Allow the 1/4 of overall error for symbol with criteria of quality evaluation between clinical examination centre chamber of the Ministry of Public Health
It closes and requires.
2.3 betweenrun precisions are tested：Daily normal value quality-control product and respectively detection 4 times of exceptional value quality-control product, add up detection 5
It, each gets 20 testing results.Calculate mean value, standard deviation, the coefficient of variation (CV%).With clinical examination centre chamber of Ministry of Public Health interstitial
Amount evaluation criterion allows the 1/3 of overall error to meet the requirements.
2.4 statistical results are shown in Table 1, from table 1 it follows that the precision verification of above-mentioned 5 representative items
As a result it meets the requirements.
The precision verification result of substrate used in 1 electrochemiluminescence analysis instrument of table
Two, measurement range is verified：
1 verification meaning：Measurement range refers to system final output value (such as concentration or activity) and tested analyte concentration
Or active proportional band.Both close to the degree of straight line, it reflected entire measurement system concentration curve for the verification of measurement range
The output characteristics of system, is very important technical indicator, directly affects the accuracy of high concentration sample results.If measured
Range is small, and the human sample Lower result of high concentration is may result in during clinical measurement, is delayed the diagnosing and treating of patient.
2 verification methods：The present embodiment according to the U.S. National Committee for Clinical Laboratory Standards file EP6-A--《It is quantitative
The linear evaluation of analysis method》, choose without haemolysis, piarhemia or jaundice etc. significantly interfere with item and the court detected close to Roche
The uppe r limit of measurement range of company's detection project and the clinical samples serum of lower limit are a case each, according to 1L, 0.8L+0.2H, 0.6L+
The volume relationship of 0.4H, 0.4L+0.6H, 0.2L+0.8H and 1H are prepared, and a series of 6 evaluations samples are formed, various concentration
Sample is analyzed in batch at one and is detected 4 times.It is X with theoretical concentration, measurement average value is Y, calculates regression equation Y=bx+a,
If correlation coefficient r >=0.975, for slope b in 0.97~1.03 range, there was no significant difference by intercept a and 0, then judges authenticator
It closes and requires.Above-mentioned each project is separately verified into its measurement range, statistical result is as shown in table 2, result of itemizing such as table 3-7 institutes
Show.
The measurement range verification result of substrate used in 2 electrochemiluminescence analysis instrument of table
3 TSH measured values of table are verified
4 E2 measurement ranges of table are verified
5 PROG measurement ranges of table are verified
6 AFP measurement ranges of table are verified
7 tPSA measurement ranges of table are verified
Shown in above table verify data and Fig. 1-5, substrate measurement range of the invention reaches the original-pack substrate of Roche
Measurement range, when sample for detecting high concentration remains to access accurately as a result, provide correct diagnostic message to clinical, is
The diagnosis and treatment of patient provide safeguard.
Three, Accuracy Verification：
1 verification method：According to National Committee for Clinical Laboratory Standards (abbreviation NCCLS) the file EP9-A2-- in the U.S.
《The bias for carrying out methods comparison with patient's sample and assessing between quantitative analysis method compares analysis》, choose fresh patient's mark
This, the requirement of table is suggested in concentration selection according to method contrast test data distribution in EP9-A2 files, covers detection range, special
It does not cover medical science decision level attentively, and there are enough amounts to carry out repeated detection.The original-pack bottom of Roche of serum sample
The statistical analysis of object and inventive substrate measurement result programs point according to the requirement of NCCLS EP9-A2 on EXCEL
Analysis is measured with the substrate of the present invention again after measuring 40 samples with the original-pack substrate of Roche respectively, measurement result (Xij,
Yij) input X, Y row, calculate and are depicted as corresponding chart automatically.The main contents include the inspections of outlier in (1) carry out method；
(2) inspection of comparison method (X) measurement range, such as r >=0.975 (or r2 >=0.95), then it is assumed that X value ranges are suitable, straight line
The slope and intercept of regression calculation are reliable；(3) regression equation y=bx+a is calculated；(4) systematic error between computational methods：According to
Clinic needs, and the medical science decision level concentration (Xc) of above-mentioned 6 representative items is substituted into regression equation, calculates this hair
Systematic error (SE) between bright substrate measurement result (Y) and the original-pack result of Roche (X), SE=︱ Yc-Xc ︱=︱ (b-1) Xc
+ a ︱；SE%=(SE/Xc) × 100%.Allow the two of overall error according to criteria of quality evaluation between clinical examination centre chamber of the Ministry of Public Health
/ mono- (1/2TEa) is criterion, if SE%<1/2TEa is then judged as substrate result and the original-pack bottom of Roche of the present invention
Object result accuracy is with uniformity, disclosure satisfy that clinical needs.
8 THS Accuracy Verifications of table
The medical science decision level of TSH is 4.2 μ IU/ml, and it is 0.197 μ IU/ml to acquire system deviation SE at this time by table 8,
SE% is 4.68, as shown in fig. 6, less than the 1/2TEa (12.5%) of the project, is judged as the substrate result and sieve of the present embodiment
The original-pack substrate result accuracy of family name is with uniformity, disclosure satisfy that clinical needs.
9 E2 Accuracy Verifications of table
The medical science decision level of E2 be 607pmol/L, at this time system deviation SE be 32.338pmol/L, SE% 5.33,
As shown in fig. 7, less than the 1/2TEa (12.5%) of the project, it is judged as the substrate result and the original-pack substrate result of Roche of the present invention
Accuracy is with uniformity, disclosure satisfy that clinical needs.
10 PROG Accuracy Verifications of table
The medical science decision level of PROG be 4.7nmol/L, at this time system deviation SE be 0.395nmol/L, SE% 8.41,
As shown in figure 8, less than the 1/2TEa (12.5%) of the project, it is judged as the substrate result and the original-pack substrate result of Roche of the present invention
Accuracy is with uniformity, disclosure satisfy that clinical needs.
11 AFP Accuracy Verifications of table
The medical science decision level of AFP is 7ng/ml, and system deviation SE is 0.488ng/ml, SE% 6.97 at this time, is such as schemed
Shown in 9, it is less than the 1/2TEa (12.5%) of the project, is judged as that the substrate result of the present invention and the original-pack substrate result of Roche are accurate
Property is with uniformity, disclosure satisfy that clinical needs.
12 tPSA Accuracy Verifications of table
The medical science decision level of tPSA is 4ng/ml, and system deviation SE is 0.186ng/ml, SE% 4.65 at this time, is such as schemed
Shown in 10, it is less than the 1/2TEa (12.5%) of the project, is judged as that the substrate result of the present invention and the original-pack substrate result of Roche are accurate
True property is with uniformity, disclosure satisfy that clinical needs.
From above-mentioned 5 representative items Accuracy Verification interpretation of result, precision of analysis can reach requirement, high
Concentration human sample result is also accurate, avoids since result caused by the low possibility of tripropyl amine (TPA) concentration is inaccurate, shows this
Invention substrate accuracy is precisely reliable.
Four, the preservation of substrate and stability verification：By luminescence buffer, the measuring cell cleaning solution and pre- in embodiment substrate
Cleaning solution, after being placed at room temperature for 24 months, each component content is unchanged, and appearance still keeps water white transparency, no precipitation, nothing to go mouldy,
The sampled above-mentioned each index of verification still conforms to provide.Show Electrogenerated chemiluminescent immunoassay system of the present invention luminescence buffer,
The stability of measuring cell cleaning solution and prerinse liquid was up to 24 months or more.
Comprehensive analysis is placed on Roche electrochemiluminescence analysis instrument and directly uses using the substrate of the present invention without calibration,
Detection range with it is original-pack consistent, hook effect will not be generated by reaching the human sample of the high concentration of upper limit of detection concentration, as a result
Accurately and reliably, so as to avoid failing to pinpoint a disease in diagnosis, there is important clinical value；24 months stationary phases, cost reduce by 60% than original-pack,
Have a vast market value.
Although the present invention has been described in detail, it will be understood by those skilled in the art that in spirit and scope of the invention
Modification will be apparent.However, it should be understood that various aspects, different specific implementation mode that the present invention records
Each section and the various features enumerated can be combined or all or part of exchange.In above-mentioned each specific implementation mode, that
A little embodiments with reference to another embodiment can be combined suitably with other embodiment, this is will be by this field skill
Art personnel are to understand.In addition, it will be understood to those of skill in the art that the description of front is only exemplary mode, not purport
In the limitation present invention.
Claims (10)
1. substrate used in a kind of electrochemiluminescence analysis instrument, which is characterized in that including luminescence buffer, measuring cell cleaning solution and pre-
Cleaning solution, luminescence buffer include that the degree of polymerization is the isooctanol polyoxy vinethene of (12-16), the bay that the degree of polymerization is (6-10)
Alcohol polyoxyethylene ether.
2. substrate according to claim 1, which is characterized in that luminescence buffer includes that the degree of polymerization is the different pungent of (12-16)
Alcohol polyoxy vinethene, the degree of polymerization be the ethoxylated dodecyl alcohol of (6-10), the concentrated phosphoric acid of 85wt%, 2 centinormal 1 dilute
Hydrochloric acid, dichloro acetamide, potassium dihydrogen phosphate and tripropyl amine (TPA).
3. substrate according to claim 2, which is characterized in that the luminescence buffer contains：
Phosphoric acid (2-5) g/L of 85wt%,
2 centinormal 1 hydrochloric acid (0.1-1.0) g/L,
Dichloro acetamide 1-3g/L,
The degree of polymerization is isooctanol polyoxy vinethene (0.6-1.2) g/L of (12-16),
The degree of polymerization is ethoxylated dodecyl alcohol (0.8-1.5) g/L of (6-10),
Potassium dihydrogen phosphate (35-40) g/L, and
Tripropyl amine (TPA) (20-30) g/L.
4. substrate according to claim 1, which is characterized in that measuring cell cleaning solution includes that the degree of polymerization is the different of (12-16)
Octanol polyoxy vinethene, the ethoxylated dodecyl alcohol and 85wt% potassium hydroxide aqueous solutions that the degree of polymerization is (6-10).
5. substrate according to claim 4, which is characterized in that the measuring cell cleaning solution contains：
The degree of polymerization is isooctanol polyoxy vinethene (0.8-1.2) g/L of (12-16),
The degree of polymerization is ethoxylated dodecyl alcohol (8-12) g/L of (6-10), and
Potassium hydroxide aqueous solution (8-12) g/L of 85wt%.
6. substrate according to claim 1, which is characterized in that prerinse liquid is phosphate buffer, contains 85wt%
Phosphoric acid, dichloro acetamide, the isooctanol polyoxy vinethene that the degree of polymerization is (12-16) and laruyl alcohol that the degree of polymerization is (6-10)
Polyoxyethylene ether.
7. substrate according to claim 6, which is characterized in that the prerinse liquid contains
Sodium dihydrogen phosphate dihydrate (0.5-1.0) g/L,
Disodium hydrogen phosphate (2.0-2.5) g/L,
Sodium chloride (1.0-1.5) g/L,
Dichloro acetamide (1.0-2.0) g/L,
85wt% phosphoric acid (180-220) mg/L,
The degree of polymerization is isooctanol polyoxy vinethene (0.8-1.2) g/L of (12-16), and
The degree of polymerization is ethoxylated dodecyl alcohol (0.8-1.2) g/L of (6-10).
8. the preparation method of substrate, feature used in a kind of electrochemiluminescence analysis instrument as described in any one of claim 1-7
It is, includes the following steps：1. preparing luminescence buffer：Each required component in addition to tripropyl amine (TPA) is weighed according to formula, with 700 milliliters
Water mixes, stirring and dissolving；Tripropyl amine (TPA) is added, stirs 10 minutes, is settled to 1000 milliliters, mixing；2. preparing measuring cell cleaning solution：
Each required component is weighed according to formula, is mixed with 700 milliliters of water, stirring and dissolving is settled to 1000 milliliters, mixing；3. preparing pre-
Cleaning solution：Each required component is weighed according to formula, is mixed with 700 milliliters of water, stirring and dissolving is settled to 1000 milliliters, mixing.
9. preparation method according to claim 8, which is characterized in that the isooctanol polyoxy vinethene degree of polymerization used is
14, the ethoxylated dodecyl alcohol degree of polymerization used is 9.
10. a kind of application of low polymerization degree surfactant in preparing substrate used in electrochemiluminescence analysis instrument, feature exist
In, the low polymerization degree surfactant include the degree of polymerization be the isooctanol polyoxy vinethene of (12-16), the degree of polymerization is (6-10)
Ethoxylated dodecyl alcohol,
Wherein, the isooctanol polyoxy vinethene degree of polymerization is preferably 14, and the ethoxylated dodecyl alcohol degree of polymerization is preferably 9.
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| CN113295678A (en) * | 2021-05-14 | 2021-08-24 | 济南迪曼生物科技有限公司 | Electrochemiluminescence cleaning fluid |
| CN115368976A (en) * | 2022-08-03 | 2022-11-22 | 吉林基蛋生物科技有限公司 | Immune lotion for electrochemiluminescence immunoassay |
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