CN108593905A - A kind of digoxin immune detection reagent and its preparation and detection method - Google Patents
A kind of digoxin immune detection reagent and its preparation and detection method Download PDFInfo
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- CN108593905A CN108593905A CN201711402527.7A CN201711402527A CN108593905A CN 108593905 A CN108593905 A CN 108593905A CN 201711402527 A CN201711402527 A CN 201711402527A CN 108593905 A CN108593905 A CN 108593905A
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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
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
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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
- G01N33/532—Production of labelled immunochemicals
- G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
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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
- G01N33/94—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors
- G01N33/9453—Cardioregulators, e.g. antihypotensives, antiarrhythmics
Abstract
The invention discloses a kind of digoxin immune detection reagent and its preparation and detection methods.Including：Enzyme mark digoxin, the indicator for detecting DigiTAb enzyme mark digoxin compound；Above-mentioned enzyme mark digoxin is coupled by digoxin and glucose dehydrogenase.The digoxin immune detection reagent of the present invention can accurately and quickly determine digoxin content in the samples such as blood of human body.Compared with existing detection reagent in the market, detection reagent of the present invention has many advantages, such as fast and easy, high sensitivity, high specificity, quantitative accurate, is conducive to clinical promote the use of.
Description
Technical field
The present invention relates to field of biological detection, specifically a kind of digoxin immune detection reagent and its preparation and detection side
Method.
Background technology
Digoxin (Digoxin, hereinafter referred to as Dig) is a kind of middle effect cardiac glycosides medicine extracted from digitalis
Object is white crystals or crystalline powder, odorless, bitter.Dig can enhance myocardial contractive power, increase heart discharge and do not increase
Add myocardial oxygen consumption, is mainly used to treat various acute and chronic cardiac insufficiencies and paroxysmal supraventricular tachycardia, atrial fibrillation, room are flutterred
The diseases such as arrhythmia cordis.The absorption of Dig is incomplete, and can be influenced by renal function with a variety of drug interactions,
Safety coefficient is small, and the concentration in blood can be because of the different difference several times of individual bioavilability.Digoxin is measured for supervising
The digoxin level of patient is surveyed to obtain best therapy, it can also be used to diagnose potential overdose.
The common method of detection digoxin has at present：Enzyme linked immunosorbent assay (ELISA) (ELISA), radiommunoassay (RIA),
Fluoroimmunoassay (FIA), chemiluminescence immunoassay (CLIA) and high efficiency liquid phase chromatographic analysis method etc..Wherein, ELISA
Standard measure accuracy is poor, the operating time is long, the degree of automation is low, is chiefly used in qualitative detection；It is longer the time required to RIA methods, detection
Unstable result, repeatability is poorer than ELISA, and there are radioactive pollution danger.FIA high specificities, sensibility is high, but needs
Expensive instrument and equipment and veteran operating personnel generally mostly use in specific medical mechanism.The reaction of CLIA methods is fast, reagent
Stability is high, but instrument and equipment and matched reagent rely on import more, and testing cost is expensive, using limited.High performance liquid chromatography
It analyzes time-consuming and is not easy to automate.
The method that the present invention uses is for homogeneous enzyme immunoassay detection method, advantage：Easy to operate, quick, high sensitivity, standard
Really property is good, is suitable for automating, and is widely used, and with automatic clinical chemistry analyzer to small-molecule substance and macromolecular substances all
It high-throughput can quickly measure.
Invention content
It is an object of the invention to solve, digoxin detection process is complicated for operation in the prior art and accuracy of measurement is low
The problem of, the present invention provides it is a kind of quickly, high sensitivity, accurately detect that the digoxin of digoxin content in sample to be tested is equal
Phase enzyme immunologic function test reagent and preparation method thereof.
To achieve the above object, the present invention provides the following technical solutions：
A kind of digoxin immune detection reagent and its preparation and detection method, it is characterised in that：Enzyme mark digoxin is used to detect ground
The indicator of digoxin antibody-enzyme mark digoxin compound；Above-mentioned enzyme mark digoxin is coupled by digoxin and glucose dehydrogenase
It forms.
As a further solution of the present invention, the indicator is selected from enzymatic reagent, including：The bottom of enzyme mark conjugate and enzyme
Object；Above-mentioned enzyme mark conjugate includes glucose dehydrogenase-digoxin conjugate；The substrate of above-mentioned enzyme is glucose.
As a further solution of the present invention, the glucose dehydrogenase-digoxin conjugate is by glucose dehydrogenase
It is coupled to be formed with digoxin.
As a further solution of the present invention, a kind of digoxin immune detection reagent and preparation method thereof, it is special
Sign is, includes the following steps：
(1) preparation of glucose dehydrogenase-digoxin conjugate：The coupling of glucose dehydrogenase (GDH) and digoxin, purifying are even
The enzyme mark digoxin of connection；
(2) preparation of digoxin homogeneous enzyme immunoassay detection reagent：
The preparation of reagent 1：It is mixed by DigiTAb and homogeneous zymolyte；
The preparation of reagent 2：It is mixed with phosphate buffer by glucose dehydrogenase-antigen conjugates.
As a further solution of the present invention, the preparation method of a kind of digoxin immune detection reagent, feature
It is, the step (1) detailed process is：
1) coupling of glucose dehydrogenase (GDH) and digoxin
A. 100-300mg digoxin is accurately weighed, 5-15mL absolute ethyl alcohols is used in combination to dissolve；
B. the sodium metaperiodate 5-15mL of 10-200mM, and slight oscillatory are added dropwise in above-mentioned a solution, reaction 0.5-2 is stirred at room temperature
Hour；
C. the ethylene glycol 0.5-1mL of 0.5-2M is added dropwise in above-mentioned b solution, reaction 5-10 minutes is stirred at room temperature；
D. above-mentioned reaction mixture is added drop-wise in the GDH solution for the 2-3% that 5-15mL is being stirred, and adjusts pH value of solution and arrives
9.0-9.5 continues to be stirred to react 0.5-2 hours, and pH value of solution is made to stablize；
E. 100-200mg tetrahydro boron sodiums, stirring reduction 12-24 hours are added in above-mentioned d solution.
2) the enzyme mark digoxin of purifying coupling
The enzyme mark digoxin being coupled by G-25 gel chromatography column purifications, obtains glucose dehydrogenase-digoxin conjugate, and in
It is stored at 2-8 DEG C.
As a further solution of the present invention, the preparation method of a kind of digoxin immune detection reagent, feature
It is, the detailed process of step (2) is as follows：
The preparation of reagent 1：By nicotinamide adenine dinucleotide NAD, the 0.5-3g glucose 0.5-2L phosphorus of 2-5g oxidation state
Homogeneous zymolyte is made in phthalate buffer dissolving；DigiTAb is added in above-mentioned homogeneous zymolyte, antibody and homogeneous enzyme bottom
The volume ratio of object is 1:100~1:10000；
The preparation of reagent 2：The glucose dehydrogenase of preparation-digoxin conjugate is added in phosphate buffer, above-mentioned coupling
The volume ratio of object and phosphate buffer is 1:100~1:10000.
As a further solution of the present invention, the detection method of the digoxin immune detection reagent, which is characterized in that
Include the following steps：
1) sample to be tested is contacted with DigiTAb；
2) according to the combination situation of enzyme mark digoxin in sample to be tested and DigiTAb, using in indicator judgement sample
The content of Gaoxin；The sample to be tested is serum, blood plasma, saliva or urine.
The principle of the present invention is that haptens is combined into enzyme mark haptens with enzyme, retains the bioactivity of haptens and enzyme, when
After enzyme mark haptens is combined with antibody, zymoprotein on hapten molecule and antibody close contact, make the activated centre of enzyme by
It influences, the activity of enzyme is suppressed.Antigen, enzyme mark haptens when measurement in sample are combined with antibody competition, in sample
Antigenic content is higher, its OD value is higher after adding substrate.
The advantage of the invention is that：The digoxin immune detection reagent of the present invention can accurately and quickly determine blood of human body
Digoxin content in equal samples.Compared with existing detection reagent in the market, detection reagent of the present invention has fast and easy, sensitive
It spends height, high specificity, quantify the advantages that accurate, be conducive to clinical promote the use of.
Description of the drawings
Fig. 1 is digoxin homogeneous enzyme immunoassay reaction calibration graph；
Fig. 2 is digoxin homogeneous enzyme immunoassay range of linearity figure；
Fig. 3 is the correlation analysis figure of the chemoluminescence method testing result of digoxin homogeneous enzyme immunoassay and NPD projects.
Specific implementation mode
The present invention provides a kind of digoxin immune detection reagent and its preparation and detection method, for make the object of the invention,
Technical solution and effect are clearer, clear, and the present invention is described in detail below.
The present invention provides a kind of digoxin immune detection reagent and its preparation and detection methods.Including：Enzyme mark digoxin,
Indicator for detecting DigiTAb-enzyme mark digoxin compound；Above-mentioned enzyme mark digoxin is by digoxin and glucose
Dehydrogenase is coupled.
Signified " digoxin " refers not only to complete digoxin molecule in the present invention, also includes that reservation intact antigen is special
The digoxin segment or derivative of property binding ability.
A kind of digoxin homogeneous enzyme immunoassay detection reagent, including：Enzyme mark digoxin, for detecting DigiTAb-enzyme mark
The indicator of digoxin compound.Indicator is selected from enzymatic reagent, radioactive isotope reagent, fluorescent reagent and chemiluminescence
Reagent.Preferably, indicator is enzymatic reagent, including：The substrate of enzyme mark conjugate and enzyme.Wherein, enzyme mark conjugate includes Portugal
Grape glucocorticoid dehydrogenase-antigen conjugates, can be obtained by chemical synthesis process.
The application method of above-mentioned digoxin immune detection reagent, includes the following steps：
1) sample to be tested is contacted with DigiTAb；
2) according to the combination situation of enzyme mark digoxin in sample to be tested and DigiTAb, using in indicator judgement sample
The content of Gaoxin；The sample to be tested is serum, blood plasma, saliva or urine etc..Preferably, sample to be tested is serum or blood plasma.
Below by specific embodiment, the present invention is described in detail.
Embodiment 1：The preparation of glucose dehydrogenase-digoxin conjugate
1) coupling of glucose dehydrogenase (GDH) and digoxin
A. 219mg digoxin is accurately weighed, 10mL absolute ethyl alcohols is used in combination to dissolve；
B. the sodium metaperiodate 10mL of 100mM, and slight oscillatory are added dropwise in above-mentioned a solution, reaction 1 hour is stirred at room temperature；
C. the ethylene glycol 0.6mL of 1M is added dropwise in above-mentioned b solution, reaction 5 minutes is stirred at room temperature；
D. above-mentioned reaction mixture is added drop-wise in the 2.8% GDH solution that 10mL is being stirred, and adjusts pH value of solution and arrives
9.0-9.5 continues to be stirred to react 1 hour, and pH value of solution is made to stablize；
E. 150mg tetrahydro boron sodiums, stirring reduction 16 hours are added in above-mentioned d solution.
2) the enzyme mark digoxin of purifying coupling
The enzyme mark digoxin being coupled by G-25 gel chromatography column purifications, obtains glucose dehydrogenase-digoxin conjugate, and in
It is stored at 2-8 DEG C.
Embodiment two：The preparation of digoxin homogeneous enzyme immunoassay detection reagent
Digoxin homogeneous enzyme immunoassay detection reagent, including：Enzyme mark digoxin, for detecting DigiTAb-enzyme mark high
The indicator of pungent compound.Indicator is selected from enzymatic reagent, radioactive isotope reagent, fluorescent reagent and chemiluminescence examination
Agent.Preferably, indicator is enzymatic reagent, including：The substrate of enzyme mark conjugate and enzyme.Wherein, enzyme mark conjugate includes grape
Glucocorticoid dehydrogenase-antigen conjugates can be obtained by chemical synthesis process.
Digoxin homogeneous enzyme immunoassay detection reagent before the use, in order to avoid the enzyme mark conjugate and enzyme in indicator
Substrate react, the substrate of enzyme mark conjugate and enzyme is separated, therefore digoxin homogeneous enzyme immunoassay detection reagent
The reagent being provided separately including two kinds, it is specific as follows：
1. the preparation of reagent 1：By nicotinamide adenine dinucleotide NAD, 1.802g of 3.588g (10mM) oxidation state
Homogeneous zymolyte is made in the phosphate buffer dissolving of (10mM) glucose 1L 50mM, 8.0 pH；DigiTAb is added to
In above-mentioned homogeneous zymolyte, the volume ratio of antibody and homogeneous zymolyte is 1:100~1:10000, in the present embodiment specifically
Ratio is 1:800.
2. the preparation of reagent 2：The phosphate that the glucose dehydrogenase of preparation-digoxin conjugate is added to 50mM, pH8.0 delays
In fliud flushing, the volume ratio of above-mentioned conjugate and phosphate buffer is 1:100~1:10000, it is specific in the present embodiment to compare
Example is 1:2000.
The application method of above-mentioned digoxin homogeneous enzyme immunoassay detection reagent, includes the following steps：
1) sample to be tested is contacted with DigiTAb；
2) according to the combination situation of enzyme mark digoxin in sample to be tested and DigiTAb, using in indicator judgement sample
The content of Gaoxin.
Specifically, sample to be tested is added in reagent 1 when detection, the digoxin in sample to be tested and the ground in reagent 1 are high
Pungent antibody is specifically bound, and generates anti digoxin antibody-digoxin compound；Reagent 2 is added, at this time in reagent 2
Glucose dehydrogenase-digoxin conjugate is mixed with the substrate of the enzyme in reagent 1, is contacted, and enzymatic reaction occurs, and constitutes detection ground
The indicator of digoxin antibody-enzyme mark digoxin compound, indicator is according to digoxin in sample to be tested and above-mentioned digoxin
The combination situation of antibody judges the content of digoxin in sample to be tested.
Since the digoxin competitive binding digoxin in glucose dehydrogenase-digoxin conjugate and sample to be tested is anti-
Body, so, the amount of digoxin is more in sample to be tested, the glucose dehydrogenase-digoxin conjugate to dissociate in homogeneous enzyme solutions
Amount it is more, enzymatic reaction is faster, leads to OD340Rise.
Above-mentioned sample to be tested is physiology sample, such as serum, blood plasma, urine, saliva etc., as a preferred solution,
Above-mentioned sample to be tested is serum or blood plasma.
Embodiment three：Digoxin homogeneous enzyme immunoassay detection reagent reacts calibration curve.
1) digoxin calibration object is prepared：Commercially available people's digoxin is dissolved in 80% ethanol solution, the school of various concentration is made
Quasi- product.Using Suzhou Bo Yuan companies digoxin calibration object as primary standard, using its calibration of digoxin kit to various concentration
Product detect 10 times respectively, find out mean value, obtain the concentration of digoxin calibration object：0.1,0.5,1.0,2.0,4.0ng/mL.
2) Biochemical Analyzer detects：By taking Hitachi 7170 operates as an example：Measurement wavelength is 340nm, takes the calibration of various concentration respectively
Digoxin R is added in product solution (15 μ L)1Reagent (200 μ L), mixing adds digoxin R2Reagent (50 μ L) after mixing, measures
The OD of different time points340Light absorption value calculates reaction rate when different calibration object concentration, needs constantly to adjust in actual mechanical process
The volume ratio of whole reagent 1 and reagent 2, while survey luminous point is adjusted, comparatively ideal reaction normal curve graph is finally obtained, per Guan Chong
Repetition measurement is 3 times fixed, and using the average value of 3 absorbance difference Δ A measured of each calibration pipe as ordinate, corresponding calibration object is a concentration of
Abscissa draws " concentration-absorbance difference " calibration curve (see Fig. 1).
Test serum or plasma sample are taken, the absorbance difference of sample is measured in the same method, substitutes into calibration curve, you can calculate
The content of digoxin in sample to be tested.If the concentration of digoxin exceeds calibration curve range in serum or blood plasma, need to be to sample
It is detected again after being diluted to ensure the accuracy of testing result.
This detection reagent is applicable not only to Hitachi 7170, applies also for semi-automatic, the full-automatic life of other brands and model
Change analyzer, design parameter can be adjusted according to instrument.
Example IV：The range of linearity determines
With the digoxin high concentration sample (3.84ng/mL) close to the range of linearity upper limit, it is pressed 1/2 with physiological saline,
1/4,1/8,1/16,1/32,1/64 dilution, is configured to the solution of 6 diluted concentrations (xi) altogether, is detected with the Biochemical Analyzer
Method measures each diluted sample concentration.Each diluted concentration is tested 3 times, finds out the mean value of each diluted concentration testing result respectively
(yi).It is that dependent variable finds out equation of linear regression to measure mean value (yi), according to formula with diluted concentration (xi) for independent variable
(1) correlation coefficient r for calculating linear regression, as a result show regression equation as y=1.0335x-0.0008, correlation coefficient r=
0.9999, show reagent of the present invention good relationship in the 0.1ng/mL-3.8ng/mL ranges of linearity (see Fig. 2).
Embodiment five：Correlation analysis
90 clinical samples including 30 positive samples and 60 ' negative ' specimens are newly produced using Shenzhen respectively
The homogeneous enzyme immunoassay method of industry biomedical engineering Co., Ltd and the present invention to sample replication 2 times, calculate separately flat respectively
Mean value carries out linear regression analysis to 90 pattern detection results, calculates the related coefficient of two kinds of reagent testing results.
The results show that correlation coefficient r=0.9964 of two kinds of kit testing results, equation of linear regression y=
0.9821x+0.0245 (see Fig. 3) shows the chemistry of digoxin homogeneous enzyme immunoassay method detection reagent and commercially available NPD projects of the present invention
Luminescence method detection reagent has good consistency.
Since the detection process of the present invention is completed by instrument full-automation, so to the of less demanding of testing staff, easily
In realizing and promote the use of.
It should be noted that obviously invention is not limited to the details of the above exemplary embodiments, the scope of the present invention is by institute
Attached claim rather than above description limit, it is intended that will fall within the meaning and scope of the equivalent requirements of the claims
All changes are included in the scope of patent protection of the present invention.
In addition, above-described is only the preferred embodiments of the invention, for the technical staff that this technology leads city,
Without departing from the principle of the present invention, several modifications and adaptations can also be done, these improved adjustment also should be regarded as this
The protection domain of invention.
Claims (7)
1. a kind of digoxin immune detection reagent and its preparation and detection method, it is characterised in that：Enzyme mark digoxin, for detecting
The indicator of DigiTAb-enzyme mark digoxin compound；
The immunologic function test reagent of digoxin according to claim 1, it is characterised in that：The enzyme mark digoxin is by digoxin
It is coupled with glucose dehydrogenase.
2. digoxin immune detection reagent according to claim 1, it is characterised in that：The indicator is tried selected from enzyme
Agent, including：The substrate of enzyme mark conjugate and enzyme；Above-mentioned enzyme mark conjugate includes glucose dehydrogenase-digoxin conjugate；It is above-mentioned
The substrate of enzyme is glucose.
3. a kind of digoxin immune detection reagent and preparation method thereof, which is characterized in that include the following steps：
（1）The preparation of glucose dehydrogenase-digoxin conjugate：Glucose dehydrogenase（GDH）With the coupling of digoxin, purifying is occasionally
The enzyme mark digoxin of connection；
（2）The preparation of digoxin homogeneous enzyme immunoassay detection reagent：
The preparation of reagent 1：It is mixed by DigiTAb and homogeneous zymolyte；
The preparation of reagent 2：It is mixed with phosphate buffer by glucose dehydrogenase-digoxin conjugate.
4. a kind of preparation method of digoxin immune detection reagent according to claim 5, which is characterized in that the step
Suddenly（1）Detailed process is：
1）Glucose dehydrogenase（GDH）With the coupling of digoxin
A. 100-300 mg digoxin is accurately weighed, 5-15 mL absolute ethyl alcohols is used in combination to dissolve；
B. the sodium metaperiodate 5-15 mL of 10-200 mM, and slight oscillatory are added dropwise in above-mentioned a solution, reaction is stirred at room temperature
0.5-2 hours；
C. the ethylene glycol 0.5-1 mL of 0.5-2 M are added dropwise in above-mentioned b solution, reaction 5-10 minutes is stirred at room temperature；
D. above-mentioned reaction mixture is added drop-wise in the GDH solution for the 2-3% that 5-15 mL are being stirred, and adjusts pH value of solution
To 9.0-9.5, continue to be stirred to react 0.5-2 hours, and pH value of solution is made to stablize；
E. 100-20 mg tetrahydro boron sodiums, stirring reduction 12-24 hours are added in above-mentioned d solution.
5.2）Purify the enzyme mark digoxin of coupling
The enzyme mark digoxin being coupled by G-25 gel chromatography column purifications, obtains glucose dehydrogenase-digoxin conjugate, and in
It is stored at 2-8 DEG C.
6. a kind of preparation method of digoxin immune detection reagent according to claim 5, which is characterized in that step（2）
Detailed process it is as follows：
The preparation of reagent 1：By nicotinamide adenine dinucleotide NAD, the 0.5-3 g glucose 0.5-2 L of 2-5 g oxidation state
Homogeneous zymolyte is made in phosphate buffer dissolving；DigiTAb is added in above-mentioned homogeneous zymolyte, antibody and homogeneous enzyme
The volume ratio of substrate is 1:100~1:10000；
The preparation of reagent 2：The glucose dehydrogenase of preparation-digoxin conjugate is added in phosphate buffer, above-mentioned coupling
The volume ratio of object and phosphate buffer is 1:100~1:10000.
7. utilizing the detection method of the digoxin immune detection reagent described in 4 any one of Claims 1-4, which is characterized in that
Include the following steps：
1）Sample to be tested is contacted with DigiTAb；
2）According to the combination situation of digoxin in sample to be tested and DigiTAb, digoxin in indicator judgement sample is utilized
Content；The sample to be tested is serum, blood plasma, saliva or urine.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110441538A (en) * | 2019-08-23 | 2019-11-12 | 北京丹大生物技术有限公司 | A kind of immuno-chromatographic test paper strip and its application for detecting digoxin |
| CN111487207A (en) * | 2019-01-09 | 2020-08-04 | 北京九强生物技术股份有限公司 | 6-glucose phosphate dehydrogenase mutant and application thereof in preparation of digoxin detection reagent |
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| CN101048660A (en) * | 2004-08-27 | 2007-10-03 | 灵芝国际股份有限公司 | Homogeneous enzyme immunoassay for oral fluid |
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2017
- 2017-12-22 CN CN201711402527.7A patent/CN108593905A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101048660A (en) * | 2004-08-27 | 2007-10-03 | 灵芝国际股份有限公司 | Homogeneous enzyme immunoassay for oral fluid |
Non-Patent Citations (1)
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111487207A (en) * | 2019-01-09 | 2020-08-04 | 北京九强生物技术股份有限公司 | 6-glucose phosphate dehydrogenase mutant and application thereof in preparation of digoxin detection reagent |
| CN111487207B (en) * | 2019-01-09 | 2023-07-07 | 北京九强生物技术股份有限公司 | Glucose 6-phosphate dehydrogenase mutant and application thereof in preparation of digoxin detection reagent |
| CN110441538A (en) * | 2019-08-23 | 2019-11-12 | 北京丹大生物技术有限公司 | A kind of immuno-chromatographic test paper strip and its application for detecting digoxin |
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Application publication date: 20180928 |