CN102565400A - Magnetic granule chemiluminescence kit for detecting melamine and application of magnetic granule chemiluminescence kit - Google Patents
Magnetic granule chemiluminescence kit for detecting melamine and application of magnetic granule chemiluminescence kit Download PDFInfo
- Publication number
- CN102565400A CN102565400A CN2010105891235A CN201010589123A CN102565400A CN 102565400 A CN102565400 A CN 102565400A CN 2010105891235 A CN2010105891235 A CN 2010105891235A CN 201010589123 A CN201010589123 A CN 201010589123A CN 102565400 A CN102565400 A CN 102565400A
- Authority
- CN
- China
- Prior art keywords
- melamine
- kit
- fluorescein
- add
- monoclonal antibody
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The invention relates to a magnetic granule chemiluminescence kit for detecting melamine, which comprises the following reagents: a luminous marker, a fluorescein marker, a standard substance, a quality control substance and a separation reagent. The luminous marker is melamine hapten marked by an isoluminol luminous marker; the fluorescein marker is a melamine monoclonal antibody marked by fluorescein or a fluorescein derivative; and the separation reagent is paramagnetic nano micro-beads enveloped in an anti-goat FITC (fluorescein isothiocyanate) monoclonal antibody. The invention further relates to a method adopting the kit to detect the melamine in animal derived food; and the method has higher sensitivity, specificity and detecting speed for melamine.
Description
Technical field
The present invention relates to a kind of chemiluminescence detection kit and method of testing thereof, particularly detect the magnetic granule chemoluminescence detection kit of content of melamine in dairy products, the feed equal samples.
Technical background
At present, the conventional sense method of melamine residual amount mainly contains high performance liquid chromatography (HPLC), gaschromatographic mass spectrometry method (GC-MS), LC/MS (LC-MS) and ELISA (ELISA) etc.
1, high performance liquid chromatography (HPLC) has accuracy of detection height, characteristics that false positive rate is low, and the lowest detection that detects melamine residual is limited to 2mg/kg.The major defect of HPLC method is that instrument costs an arm and a leg, operate more loaded down with trivial details, length consuming time, testing cost is expensive, and testing staff's specialty is had relatively high expectations, the sample pre-treatment is complicated.
2, gas chromatography-mass spectrography (GC-MS) sensitivity is very high, and false positive rate is low, and the lowest detection that detects melamine residual is limited to 0.05mg/Kg.The major defect of this method is that sample needs derivatization to handle complicated like this early stage.Though can in mass spectrum, provide more structural information through the sample of derivatization treatment, derivatization can produce a plurality of different products and cause the partial loss of sample, causes the deviation of experimental result.
3, LC/MS (LC-MS), sample does not need derivatization treatment, can detect urine, blood, liver, hair and eyeball sample.This method reaches 0.01mg/Kg to the LDL of melamine, and the LC-MS/MS coupling can further improve signal to noise ratio (S/N ratio), so can be used for the affirmation means to positive findings.But, no matter LC-MS or LC-MS/MS, the instrument detecting method is the same with GC-MS, and fails to solve instrument and involve great expense, complex operation step, the sample complex pretreatment requires problems such as height to operating personnel's specialty attainment.
4, ELISA (ELISA) is to occur the seventies in 20th century; Be used for the detection of micro substance; Be applied to Clinical detection such as infectious disease, tumor markers, hormonal readiness the earliest; Beginning the nineties to apply in China food safety detection field, rely on the kit of elisa technique, the leading products that test strips has become food security fast detecting field at present, also is simultaneously the major product type that my company researches and develops and sells.The enzyme linked immunosorbent detection method is based on the specific reaction of Ag-Ab; Detection sensitivity is higher, specificity is better; Technical operation is simple and easy; Easy master, qualitative, the quantitative test work that have solved a large amount of former insoluble many micro-small-molecule substances such as microbiotic, hormone, residues of pesticides etc. have really been played positive facilitation to the development of food security Fast Detection Technique.But there are many defectives that self can't overcome in the ELISA method, and mainly shows:
1) heterogeneous reaction: be separated free thing and bond in the testing process, need multistep to wash the plate process, time and effort consuming is difficult to improve the automation of operation degree.
2) enzymic catalytic reaction: by substrate for enzymatic activity colour developing, assaying reaction liquid absorbance.There are considerable influence in the time and the temperature of reaction to enzyme activity, and reagent stability is poor.
Summary of the invention
Technical matters to be solved by this invention is to provide a kind of melamine detection kit, not only possesses higher sensitivity when adopting this kit to carry out melamine detection, specificity, and have higher reaction velocity.
Another object of the present invention is to provide a kind of method of testing of melamine, and this method is simple to operate, and is highly sensitive, and specificity is good.
Realize above-mentioned purpose, the present invention provides a kind of melamine detection kit, and its main agents that comprises has: luminous marker, fluorescein-labelled thing, standard items, quality-control product, separation agent.
Described separation agent is the paramagnetism nano microsphere that is coated with goat-anti FITC monoclonal antibody.
Described paramagnetism nano microsphere, surface contain-OH ,-COOH or-NH
2The microballon of reactive group is used to encapsulate goat-anti FITC monoclonal antibody, and its inner core is Fe
3O
4Or γ-Fe
2O
3, make microballon have paramagnetism.
Described luminous marker is the melamine hapten of different luminol derivant mark.Described different luminol derivant is ABEI, AHEI or ABEN.
Described kit also comprises standard items, quality-control product and concentrated washing lotion.
Described fluorescein-labelled thing is a FITC mark melamine monoclonal antibody.
The present invention also provides a kind of method of utilizing kit to detect melamine, comprises the following steps:
1) draw 20 μ l-100 μ l standard items or samples respectively, add 20 μ l-100 μ l luminous markers then, add the fluorescein-labelled thing of 20 μ l-100 μ l again, behind the abundant mixing, 37 ℃ of incubation 15min;
2) add the paramagnetism nano microsphere 80-150 μ l be coated with goat-anti FITC monoclonal antibody, behind the mixing at 37 ℃ of incubation 5min;
3) separate 5min with the magnetic separator frame, precipitate with cleaning fluid 300-500 μ l flushing compound after abandoning supernatant;
4) above-mentioned cleaning step repeats 2-4 time;
5) 4) compound of gained separator well is directly put into the measurement camera bellows; Add and excite substrate 1 and excite substrate 2; Postpone to detect the relative light intensity (RLU) that sends behind the 3-5s, content in the sample and RLU proportion relation can be through the concentration of RLU set calibration curve method calculating melamine.
The principal ingredient of described luminous substrate is NaOH and H
2O
2
The used analyser of analysis test method is among the present invention: comprise power circuit, automatic syringe pump 1 and 2, measuring chamber, illuminated chamber, photomultiplier counter and output system; Also dispose the Windows Control Software of computing machine and Chinese interface simultaneously; Can carry out that data typing, result gather, quality control, the result stores and function such as result queries; Can accomplish the programming of multiple analytical model, quantitative or qualitative reporting the result generates and storage, update functions automatically; Automatically revise typical curve at 2, the system that is adopted is the CI-2008 system.
Separation agent of the present invention is to encapsulate goat-anti FITC monoclonal antibody, and the content of its surface group is 0.1-0.3eqm/g, and it is stored in 0.8-1.2% casein, 0.2-0.5%Tween-20,0.1-0.3%NaN
3, in the pH7.2-7.6TRIS damping fluid.Described FITC is a fluorescein isothiocynate.Said percentage composition is the quality percentage composition.
Described luminous marker is the melamine hapten of different luminol derivant ABEI, AHEI or ABEN mark, and it is stored in pH7.2-7.6, contains 0.2-0.5%Tween-20,0.1-0.3%NaN
3The TRIS damping fluid in.Said percentage composition is the quality percentage composition.
The melamine monoclonal antibody that described fluorescein-labelled thing is the FITC mark, it is stored in pH7.2-7.6, contains the 1.0-2.0% ovalbumin, 0.2-0.4%NaN
3, the 0.05-0.1mol/LTRIS damping fluid.Said percentage composition is the quality percentage composition.
The melamine standard solution (0ng/ml, 0.01ng/ml, 0.03ng/m, 0.09ng/ml, 0.27ng/ml, 0.81ng/ml), the standard items dilution is pH7.0-7.5,0.2-0.4%NaN
3, the 0.2-0.3mol/L borate buffer solution.Said percentage composition is the quality percentage composition.
Melamine quality-control product solution concentration is respectively 0.02ng/ml, and 0.5ng/ml, standard items dilution are pH7.0-7.5,0.2-0.4%NaN
3, the 0.2-0.3mol/L borate buffer solution.Said percentage composition is the quality percentage composition.
Described concentrated washing lotion is PH7.2-7.8,0.2-0.4%Tween-20,0.2-0.4%NaN
3, the 0.1-0.2mol/LPBS damping fluid.Said percentage composition is the quality percentage composition.
Beneficial effect of the present invention is following:
1) but kit specific detection melamine of the present invention does not have intersection to other drug.
2) sensitivity of kit of the present invention is higher, can reach 10ng/ml to melamine detection sensitivity.
3) detection of kit of the present invention is quick, is lower than 20min detection time.
Embodiment
The preparation of the concrete component of embodiment 1 kit
1, the preparation of luminous marker
A) haptenic synthetic
Get 0.5g (3.97mmol) melamine in the 50ml of drying round-bottomed flask; Add 35ml DMF stirring and dissolving, get 0.44g (7.93mmol) KOH, 0.12g anhydrous Na I adds in the above-mentioned solution, drip the 0.646ml bromo-acetic acid tert-butyl; After fully mixing, 65 ℃ of reactions of heat temperature raising 10h.Reaction is cooled to room temperature after finishing, and thin up is used ethyl acetate extraction, and drying is revolved steaming, gets yellow oil, ethyl acetate: sherwood oil=cross post get melamine hapten at 2: 1.
B) luminous marker preparation
Get 4.5mmol/L ABEI, be dissolved in the 4ml distilled water, the 5.0mmol/L N-hydroxy-succinamide is dissolved in 0.5ml N, in the dinethylformamide, and room temperature reaction 3-4h behind the two abundant mixing.Get the melamine hapten 15mg of above-mentioned preparation, to 1.5ml, add the ABEI solution of above-mentioned activation with the pH7.4PBS adjusted volume then, fully room temperature reaction spends the night behind the mixing, crosses G-25 gel column purifying.
2, fluorescent marker preparation
A) immunogenic preparation:
Adopt carbodiimide method to carry out coupling melamine hapten and ovalbumin and obtain immunogene.
B) melamine MONOCLONAL ANTIBODIES SPECIFIC FOR
Animal immune: with immunogene the Balb/c mouse is carried out immunity, immunizing dose is 150 μ g/, makes it produce antiserum.
Fusion of Cells and cloning: get immune Balb/c mouse boosting cell, merge, obtain the hybridoma cell strain of monoclonal antibody in 9: 1 ratios and SP2/0 myeloma cell.
Cell cryopreservation and recovery: hybridoma is processed 1 * 10 with cryopreserving liquid
9The cell suspension of individual/ml is in the medium-term and long-term preservation of liquid nitrogen.Take out frozen pipe during recovery, put into 37 ℃ of water-bath middling speeds immediately and melt, behind the centrifugal removal cryopreserving liquid, move in the culture flask and cultivate.
MONOCLONAL ANTIBODIES SPECIFIC FOR and purifying: the Balb/c mouse peritoneal is only injected sterilization paraffinum liquidum 0.5ml/, 7 days pneumoretroperitoneum injection hybridomas 5 * 10
5Individual/as only, to gather ascites after 7 days.Carry out purifying, obtain monoclonal antibody with sad-saturated ammonium sulfate method.
C) fluorescent marker preparation
Use 0.025mol/L, the carbonate buffer solution of pH9.0 is diluted to 1% mass percent concentration with the melamine monoclonal antibody; Total protein according to desiring mark adds 0.01mg FITC by every milligram of immunoglobulin (Ig), accurately takes by weighing required FITC powder with analytical balance.With the solution that FITC is made into 0.1mg/ml,, sneak into the FITC dilution with same damping fluid by 3-5 times of above-mentioned antibody-solutions volume; Electromagnetic agitation, mark 30-48h under 4～℃ lucifuge condition; With label solution 3000r/min, the centrifugal 20min of room temperature removes wherein a spot of sediment, and in the bag filter of packing into, the PBS damping fluid of pH7.4 dialysis is 2-3 days again, during change dislysate at least 3 times; Get the label of dialysed overnight, through SephadexG-25 or G-50 post, the separated free luciferin is collected the fluorescent marker of mark and is identified that packing is stored in 4 ℃ of refrigerators.
3, separation agent preparation
A) magnetic bead activation
The magnetic bead of surface-COOH group (purchase in DYNAL, particle diameter is 2.8 μ m), its content is 0.15eq/g; Get 100 μ l magnetic beads, with 100 μ l 25mmol/L, pH5.0,0.05%Tween-20MES solution washing twice, magnetic removes supernatant after separating; Before the use, dispose EDC, the NHS solution of 50mmol/L respectively with the 25mmol/L MES solution of 4 ℃ of storages; Add new EDC that disposes of 50 μ l and NHS solution respectively in the centrifuge tube that magnetic bead is housed, vortex mixing, room temperature activation 30min; Centrifuge tube places on the magnetic separator frame magnetic to separate 4min, removes supernatant, adds 100 μ l, 25mmol/L, pH5.0, MES clean get final product after 2-3 time the magnetic bead of surperficial activated carboxylic.
B) magnetic bead coupling goat-anti FITC monoclonal antibody
50-100 μ g goat-anti FITC monoclonal antibody is dissolved into 60 μ l, and 25mmol/L among the pH5.0MES, regulates cumulative volume to 100 μ l with said MES solution, soft mixing magnetic bead and antibody; At least coupling 30min or 4 ℃ of coupling 2h under the room temperature condition, vortex appearance capable of using makes magnetic bead keep the mixing state during this period; Centrifuge tube places magnetic separation 3-5min on the magnetic separator frame, removes supernatant; For cancellation unreacted-COOH, can add 100 μ l, pH7.4, TRIS reaction 15min or 100 μ l, pH8.0 contains the PBS sealing magnetic bead of 50mmol/L monoethanolamine; With 100 μ l, 0.1-0.3%BSA, the PBS of 0.1%Tween-20 or TRIS clean the good magnetic bead of sealing 3-5 time; At last magnetic bead is redissolved in containing 0.1-0.5%BSA, 0.01-0.1%Tween-20,0.02%NaN
3PBS or TRIS damping fluid in, 2-8 ℃ of preservation.
The establishment of embodiment two kits
Set up the magnetic granule chemoluminescence detection kit that detects melamine, make it contain following component:
The fluorescent marker of the melamine monoclonal antibody of FITC mark
The luminous marker of the melamine hapten of ABEI mark
The separation agent of the paramagnetism Nano microsphere of the anti-sheep FITC of pan coating monoclonal antibody
The melamine standard solution (0ng/ml, 0.01ng/ml, 0.03ng/m, 0.09ng/ml, 0.27ng/ml, 0.81ng/ml), the standard items dilution is pH7.2,0.3%NaN
3, the 0.3mol/L borate buffer solution.Said percentage composition is the quality percentage composition.
Melamine quality-control product solution concentration is respectively 0.02ng/ml, and 0.5ng/ml, standard items dilution are pH7.5,0.3%NaN
3, the 0.3mol/L borate buffer solution.Said percentage composition is the quality percentage composition.
Concentrated washing lotion is PH7.4,0.3%Tween-20,0.4%NaN
3, 0.2mol/L PBS damping fluid.Said percentage composition is the quality percentage composition.
Melamine detection in embodiment three actual samples
1, sample pre-treatment
(1) milk sample pre-treating method:
Get 400 μ l fresh milk samples; Add 800 μ l acetonitriles, whirling motion 1min gets 100 μ l supernatants and adds 500 μ l redissolution working fluid, mixing; Getting 50 μ l is used for analyzing diluted sample multiple: 20.
(2) milk powder sample pre-treating method:
Take by weighing 1.0 ± 0.05g milk powder sample to the 10ml centrifuge tube, add the 2.5ml deionized water, whirling motion 1min; Pipette 400 μ l in the 2ml centrifuge tube, add 400 μ l acetonitriles, add 400 μ l methyl alcohol again, whirling motion 1min, more than the 3000g, the centrifugal 3-5min of room temperature (20-25 ℃); Get 100 μ l supernatants and add 500 μ l redissolution working fluid, mixing; Getting 50 μ l is used for analyzing diluted sample multiple: 20
(3) feed sample pre-treating method:
Take by weighing in feed sample to the 50ml polystyrene centrifuge tube of 2.0 ± 0.05g pulverizing; Add 9ml methyl alcohol and add 1ml again and extract working fluid, with the oscillator 5min that vibrates; More than the 3000g, the centrifugal 10min of room temperature (20-25 ℃); Add 950 μ l and redissolve working fluid in melamine purification centrifuge tube, pipette 50 μ l feed supernatants again and add wherein, whirling motion 3min, fully mixing; More than the 3000g, the centrifugal 10min of room temperature (20-25 ℃); Getting supernatant liquor 50 μ l is used for analyzing diluted sample multiple: 100.
2, detect and interpretation of result with kit
Draw 20 μ l-100 μ l standard items or samples respectively, add 20 μ l-100 μ l luminous markers then, add the fluorescein-labelled thing of 20 μ l-100 μ l again, behind the abundant mixing, 37 ℃ of incubation 15min; Add the paramagnetism Nano microsphere 80-150 μ l be coated with goat-anti FITC monoclonal antibody, behind the mixing at 37 ℃ of incubation 5min; Separate 5min with the magnetic separator frame, precipitate with cleaning fluid 300-500 μ l flushing compound after abandoning supernatant; The compound of gained separator well is directly put into the measurement camera bellows; Add and excite substrate 1 and excite substrate 2; Postpone to detect the relative light intensity (RLU) that sends behind the 3-5s; The content of melamine and RLU proportion relation in the sample can be through the concentration of RLU combined standard curve method calculating melamine.
Exciting substrate 1 is NaOH, and exciting substrate 2 is H
2O
2Before substrate loads, clean the substrate pump 10-20 time, behind the remaining water mark, more corresponding substrate is directly put into instrument in the emptying pipe, pump line is inserted in the substrate bottle with distilled water; With substrate flushing pipe 5 times, and measure the RLU value of substrate, under the normal condition, the RLU value of substrate should be above 1200.If surpass 1200, need with distilled water pipeline and substrate pump to be carried out more times cleaning again, in blank value is reduced to zone of reasonableness.Do not do experiment if surpass three days, need unloading substrate bottle, and cover lid, with vaporization prevention.Use the distilled water pipe blow-through subsequently, and emptying, in order to avoid strong base solution corrosion substrate pump.
The present invention adopt 6 melamine standard items (0ng/ml, 0.01ng/ml, 0.03ng/m, 0.09ng/ml, 0.27ng/ml 0.81ng/ml) carries out plotting curves.Instrument detects according to said method and obtains a calibration curve that the RLU value is relevant with melamine concentration, and in after this measuring, the melamine concentration in each sample is compared with typical curve and drawn the content of melamine in the sample.
The mensuration of embodiment four kit quality
1, the sensitivity of kit
Being defined as of kit sensitivity: measure 20 times the zero standard article, the mean value of mensuration adds 3 times of standard deviations.The sensitivity of this kit is 0.01ng/ml.
2, the accuracy of sample and precision
Accuracy is meant the matching degree between measured value and true value, and the kit accuracy recovery commonly used is represented.Precision is claimed repeatability again, and the coefficient of variation commonly used is represented.
Sample extraction method according to embodiment three; Melamine with 0.5ng/g (ml), two concentration of 1ng/g (ml) adds recovery to milk, milk powder sample respectively; Melamine with 2ng/g, two concentration of 4ng/g adds recovery to the feed sample respectively; Every kind of each 4 of each concentration of sample are parallel, measure with three batches of kits, calculate the average recovery rate and the precision of sample.Experimental result sees the following form.
Table 1 accuracy and precision are measured ng/g (ml)
Can know from table, the average recovery rate scope that two concentration of melamine are all added in milk, milk powder, the feed sample between 88.7-98.9%, in batch, batch between all the coefficient of variation less than 15%.
3, specificity
Cross reacting rate is meant the ability that antibody combines with structure different antigens determinant.
Table 2 kit cross reacting rate
| Medicine name | Cross reacting rate |
| Melamine | 100％ |
| Cyanuric acid | ＜1％ |
| Triazine | ＜1％ |
| Triazinediamine | ＜1％ |
4, correlativity
X＝CI-2008，Y＝RIA
Y＝1.25X-2.96
R＝0.9752
X is CI-2008 system measurement result, and Y is that the result is measured in the aspect
Claims (8)
1. a magnetic particle that detects melamine is competed chemical luminescence reagent kit, and the reagent that comprises has: luminous marker, fluorescein-labelled thing, standard items, quality-control product, separation agent.
2. kit according to claim 1 is characterized in that: described separation agent is the paramagnetism nano microsphere that is coated with goat-anti FITC monoclonal antibody.
3. kit according to claim 2 is characterized in that: described paramagnetism nano microsphere is that the inside is coated with Fe
3O
4Or γ-Fe
2O
3, the surface contains-OH ,-COOH or-NH
2The microballon of reactive group.
4. according to each described kit of claim 1-3, it is characterized in that: described luminous marker is the melamine hapten of different luminol derivant mark.
5. kit according to claim 4 is characterized in that: described different luminol luminous marker is ABEI, AHEI or ABEN.
6. according to each described kit of claim 1-3, it is characterized in that: described kit also comprises titer, quality-control product and concentrated washing lotion.
7. according to the described kit of one of claim 1-3, it is characterized in that: the melamine monoclonal antibody that described fluorescein-labelled thing is the FITC mark.
8. a method of utilizing each described kit of claim 1-7 to detect melamine comprises the following steps:
1) draw 20 μ l-100 μ l standard items or samples respectively, add 20 μ l-100 μ l luminous markers then, add the fluorescein-labelled thing of 20 μ l-100 μ l again, behind the abundant mixing, 37 ℃ of incubation 15min;
2) add the paramagnetism nano microsphere 80-150 μ l be coated with goat-anti FITC monoclonal antibody, behind the mixing at 37 ℃ of incubation 5min;
3) separate 5min with the magnetic separator frame, precipitate with cleaning fluid 300-500 μ l flushing compound after abandoning supernatant;
4) above-mentioned cleaning step repeats 2-4 time;
5) 4) compound of gained separator well is directly put into the measurement camera bellows; Add and excite substrate 1 and excite substrate 2; Postpone to detect the relative light intensity (RLU) that sends behind the 3-5s; The content of melamine and RLU proportion relation in the sample can be through the concentration of RLU set calibration curve method calculating melamine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105891235A CN102565400A (en) | 2010-12-15 | 2010-12-15 | Magnetic granule chemiluminescence kit for detecting melamine and application of magnetic granule chemiluminescence kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105891235A CN102565400A (en) | 2010-12-15 | 2010-12-15 | Magnetic granule chemiluminescence kit for detecting melamine and application of magnetic granule chemiluminescence kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102565400A true CN102565400A (en) | 2012-07-11 |
Family
ID=46411357
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010105891235A Pending CN102565400A (en) | 2010-12-15 | 2010-12-15 | Magnetic granule chemiluminescence kit for detecting melamine and application of magnetic granule chemiluminescence kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102565400A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103439320A (en) * | 2013-09-04 | 2013-12-11 | 青岛科技大学 | Method for determining melamine (Me) by chemiluminescence |
| CN105181680A (en) * | 2015-09-28 | 2015-12-23 | 成都博奥新景医学科技有限公司 | Magnetic bead separation chemiluminescence immunoassay method for melamine |
| CN107490574A (en) * | 2017-08-11 | 2017-12-19 | 太原瑞盛生物科技有限公司 | A kind of chemiluminescence detection kit of melamine and preparation method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1888901A (en) * | 2006-04-21 | 2007-01-03 | 深圳市新产业生物医学工程有限公司 | Magnetic separating direct chemical illuminating reagent and testing method using the same reagent |
| CN101324578A (en) * | 2007-06-13 | 2008-12-17 | 清华大学 | Immune radiometric analysis reagent kit for detecting tumour sign object CA50 and use method thereof |
| CN101413943A (en) * | 2008-12-02 | 2009-04-22 | 北京望尔康泰生物技术有限公司 | Method for detecting melamine and specific enzyme-linked immunologic reagent kit |
| CN101424685A (en) * | 2008-11-14 | 2009-05-06 | 深圳市绿诗源生物技术有限公司 | ELISA kit for detecting melamine and method |
| CN201298041Y (en) * | 2008-11-12 | 2009-08-26 | 河南省农业科学院 | Rapid detecting test paper for melamine |
| CN101688863A (en) * | 2007-04-23 | 2010-03-31 | 斯莱特抗体有限公司 | Assay devices and methods and components for use therein |
-
2010
- 2010-12-15 CN CN2010105891235A patent/CN102565400A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1888901A (en) * | 2006-04-21 | 2007-01-03 | 深圳市新产业生物医学工程有限公司 | Magnetic separating direct chemical illuminating reagent and testing method using the same reagent |
| CN101688863A (en) * | 2007-04-23 | 2010-03-31 | 斯莱特抗体有限公司 | Assay devices and methods and components for use therein |
| CN101324578A (en) * | 2007-06-13 | 2008-12-17 | 清华大学 | Immune radiometric analysis reagent kit for detecting tumour sign object CA50 and use method thereof |
| CN201298041Y (en) * | 2008-11-12 | 2009-08-26 | 河南省农业科学院 | Rapid detecting test paper for melamine |
| CN101424685A (en) * | 2008-11-14 | 2009-05-06 | 深圳市绿诗源生物技术有限公司 | ELISA kit for detecting melamine and method |
| CN101413943A (en) * | 2008-12-02 | 2009-04-22 | 北京望尔康泰生物技术有限公司 | Method for detecting melamine and specific enzyme-linked immunologic reagent kit |
Non-Patent Citations (9)
| Title |
|---|
| ERIC A.E.GARBER: "Detection of Melamine Using Commercial Enzyme-Linked Immunosorbent Assay Technology", 《JOURNAL OF FOOD PROTECTION》 * |
| JOOHEE CHOI,ET AL.: "Rapid quantification of melamine in milk using competitve 1,1`-oxalyldiimidazole chemiluminescent enzyme immunoassy", 《ANALYST》 * |
| TSUNG-HSUAN TSAI,ET AL.: "Detection of Melamine in Milk Powder and Human Urine", 《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》 * |
| ZHUMING WANG,ET AL.: "Subpicogram determination of melamine in milk products using a luminol-myoglobin chemilumiescence system", 《AGRICULTURAL AND FOOD CHEMISTRY》 * |
| 尹东光: "几种主要化学发光物质的发光性能及其化学发光免疫分析体系", 《标记免疫分析与临床》 * |
| 林祥梅，王建峰，贾广乐等: "三聚氰胺的毒性研究", 《毒理学杂志》 * |
| 柳其芳: "ELISA法测定三聚氰胺效果分析", 《中国热带医学》 * |
| 王栩 等: "化学发光免疫分析技术新进展", 《分析试验室》 * |
| 高红军 等: "化学发光免疫分析法检测血清NSE的室间比对结果分析", 《中国卫生检验杂志》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103439320A (en) * | 2013-09-04 | 2013-12-11 | 青岛科技大学 | Method for determining melamine (Me) by chemiluminescence |
| CN103439320B (en) * | 2013-09-04 | 2015-06-17 | 青岛科技大学 | Method for determining melamine (Me) by chemiluminescence |
| CN105181680A (en) * | 2015-09-28 | 2015-12-23 | 成都博奥新景医学科技有限公司 | Magnetic bead separation chemiluminescence immunoassay method for melamine |
| CN105181680B (en) * | 2015-09-28 | 2018-05-04 | 成都博奥新景医学科技有限公司 | Magnetic bead separation chemiluminescence immunoassay method for melamine |
| CN107490574A (en) * | 2017-08-11 | 2017-12-19 | 太原瑞盛生物科技有限公司 | A kind of chemiluminescence detection kit of melamine and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101226194B (en) | Malachite green vestigial ELISA detection kit and usage method thereof | |
| CN101571539B (en) | Elisa kit for detecting cephalo-type medicine and application thereof | |
| CN102539412A (en) | Magnetic particle chemical luminous kit for detecting clenbuterol and application thereof | |
| CN102928413A (en) | Magnetic particle chemiluminescence kit for detecting tetracyclines, and applications thereof | |
| CN102928417A (en) | Magnetic particle chemiluminescence kit for detecting sulfanilamide drugs, and applications thereof | |
| CN102608316B (en) | Kit or test strip for detecting quinoxaline compound | |
| CN102928410A (en) | Magnetic particle chemiluminescence kit for detecting ractopamin, and applications thereof | |
| CN102565400A (en) | Magnetic granule chemiluminescence kit for detecting melamine and application of magnetic granule chemiluminescence kit | |
| CN102368054A (en) | Chemiluminescence ELISA kit for detecting concentration of drug tacrolimus in human whole blood | |
| CN102928407A (en) | Magnetic particle chemiluminescence kit for detecting avermectins, and applications thereof | |
| CN101935348A (en) | Lead ion antigen and preparation method and application thereof | |
| CN102928411A (en) | Magnetic particle chemiluminescence kit for detecting streptomycin, and applications thereof | |
| CN102928406A (en) | Magnetic particle chemiluminescence kit for detecting sudan I, and applications thereof | |
| CN102539413A (en) | Magnetic particle chemiluminescence kit for detecting frazolidone metabolites and application of magnetic particle chemiluminescence kit | |
| CN103592435A (en) | Magnetic-particle chemiluminescence kit used for detecting monensin and applications of the kit | |
| CN102928409A (en) | Magnetic particle chemiluminescence kit for detecting lincomycin, and applications thereof | |
| CN102565032A (en) | Magnetic granule chemiluminescence kit for detecting nitrofurantoin metabolite and application of magnetic granule chemiluminescence kit | |
| CN102928592A (en) | Magnetic particle chemiluminescence kit for detecting trenbolone, and applications thereof | |
| CN102539762B (en) | Enzyme-linked immunosorbent assay kit for detecting Sudan red and paranitroaniline red medicaments and application thereof | |
| CN102539763A (en) | Magnetic particle chemiluminescent kit for detecting furaltadone metabolite and application thereof | |
| CN102230936A (en) | Immunochromatography test paper for detecting ractopamine and preparation method thereof | |
| CN102928405A (en) | Magnetic particle chemiluminescence kit for detecting nitroimidazoles, and applications thereof | |
| CN202631541U (en) | Colloidal gold kit for detecting fluoroquinolones medicines in milk | |
| CN102928415A (en) | Magnetic particle chemiluminescence kit for detecting estradiol, and applications thereof | |
| CN102565403A (en) | Magnetic particle chemiluminescence kit for detecting furaltadone and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20120711 |