CN102539414A - Magnetic particle chemiluminescence kit for detecting nitrofurantion and application thereof - Google Patents
Magnetic particle chemiluminescence kit for detecting nitrofurantion and application thereof Download PDFInfo
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- CN102539414A CN102539414A CN2010105929251A CN201010592925A CN102539414A CN 102539414 A CN102539414 A CN 102539414A CN 2010105929251 A CN2010105929251 A CN 2010105929251A CN 201010592925 A CN201010592925 A CN 201010592925A CN 102539414 A CN102539414 A CN 102539414A
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Abstract
The invention relates to a magnetic particle chemiluminescence kit for detecting nitrofurantion. The kit comprises the following reagents: a luminescent label, a fluorescent label, a standard product, a quality control product and a separating reagent. The luminescent label is of a nitrofurantion hapten labeled by an isoluminol luminescent label, the fluorescent label is of a nitrofurantion monoclonal antibody labeled by fluorescein or derivatives thereof, and the separating reagent is of paramagnetic nano micro-beads coated with a goat anti-FITC (fluorescein isothiocyanate) monoclonal antibody. The invention further relates to a method for detecting the nitrofurantion in feed by adopting the kit, and the method has the advantages of higher sensitivity in detection of the nitrofurantion, higher specificity and faster detection speed.
Description
Technical field
The present invention relates to a kind of direct chemical luminous detection kit and method of testing thereof, particularly detect the magnetic particle competition direct chemical luminous detection kit of Nitrofurantoin residue amount in the feed sample.
Technical background
The itrofurans medicine is the broad-spectrum antibiotic of synthetic, and it has extraordinary antibacterial action and the dynamic (dynamical) characteristic of medicine, once is widely used as domestic animal, poultry, propagates the growth-promoting additive of aquatic products artificially.But find through long-term experimental study; Itrofurans medicine and metabolin all can make animal used as test that canceration and gene mutation take place; And infer thus; Human long-term use such medicine or long-term edible poultry, domestic animal of adding such growth-promoting additive, propagate aquatic products artificially, equally also canceration and gene mutation can take place.So this type of drug withdrawal uses in treatment and feed.
At present, the conventional sense method of Nitrofurantoin residue amount mainly contains high performance liquid chromatography (HPLC), Ultra Performance Liquid Chromatography method (UPLC) and enzyme linked immunosorbent assay (ELISA) etc.
1, high performance liquid chromatography (HPLC) has accuracy of detection height, characteristics that false positive rate is low.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, Ultra Performance Liquid Chromatography method (UPLC) is far superior to liquid phase chromatography on analysis time and solvent load, demonstrates its good superiority when especially the complicated trace components of matrix being measured.Even like this, UPLC fails to solve instrument and involves great expense, complex operation step, and the sample complex pretreatment requires problems such as height to operating personnel's specialty attainment.
3, enzyme linked immunosorbent assay (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 furantoin detection kit, not only possesses higher sensitivity when adopting this kit to carry out the detection of furantoin, specificity, and have higher reaction velocity.
Another object of the present invention is to provide a kind of method of testing of furantoin, 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 furantoin 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 furantoin haptens 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.
The furantoin monoclonal antibody that described fluorescein-labelled thing is the FITC mark.
The present invention also provides a kind of method of utilizing kit to detect furantoin, 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 furantoin.
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 pH7.1-7.5; Contain the 3-5% casein; The 0.1-0.3% Tween-20,0.02-0.04% thimerosal antiseptic is in the 0.03-0.05mol/LTRIS damping fluid.Described FITC is a fluorescein isothiocynate.Said percentage composition is the quality percentage composition.
Described luminous marker is the furantoin haptens of different luminol derivant ABEI, AHEI or ABEN mark, and it is stored in it and is stored in pH7.2-7.6, contains the 0.1-0.3% Tween-20,0.02-0.04%NaN
3The TRIS damping fluid in.Said percentage composition is the quality percentage composition.
The furantoin monoclonal antibody that described fluorescein-labelled thing is the FITC mark, it is stored in pH7.2-7.6, contains the 8-10% ovalbumin, 0.02-0.04% thimerosal antiseptic, 0.1-0.2mol/LPBS damping fluid.Said percentage composition is the quality percentage composition.
The furantoin standard solution (0ng/ml, 0.01ng/ml, 0.03ng/ml, 0.09ng/ml, 0.27ng/ml, 0.81ng/ml), the standard items dilution is pH7.2-7.6,0.03-0.05% thimerosal antiseptic, 0.05-0.1mol/LTRIS damping fluid.Said percentage composition is the quality percentage composition.
Furantoin quality-control product solution concentration is respectively 0.02ng/ml, 0.5ng/ml, quality-control product dilution pH7.2-7.4,0.03-0.05% thimerosal antiseptic, 0.05-0.1mol/L TRIS damping fluid.Said percentage composition is the quality percentage composition
Described concentrated washing lotion is PH7.0-7.4,0.2-0.4% Tween-20,0.2-0.4% sodium azide, 0.1-0.2mol/L PBS damping fluid.Said percentage composition is the quality percentage composition.
Beneficial effect of the present invention is following:
1) but kit specific detection furantoin of the present invention does not have intersection to other drug.
2) sensitivity of kit of the present invention is higher, can reach 0.01ng/ml to the detection sensitivity of furantoin.
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
React in water with furantoin with to carboxyl benzaldehyde and to carry out derivatization and obtain the furantoin derivative hapten.
Getting furantoin 500mg is dissolved in the 5ml pure water.500mg is dissolved in the 15ml water carboxyl benzaldehyde, adds room temperature reaction 48h in the furantoin solution, obtain faint yellow deposition and be the furantoin derivative hapten.Clean 5-6 time repeatedly with 25ml water, obtain the furantoin derivative hapten after the oven dry.
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 furantoin haptens 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 mixed anhydride method to carry out coupling furantoin haptens and ovalbumin and obtain immunogene.
B) furantoin 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
6The 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 with sad-saturated ammonium sulfate method, the ascites behind the purifying is put into-20 ℃ of environment and is preserved.
C) fluorescent marker preparation
Use 0.025mol/L, the carbonate buffer solution of pH9.0 is diluted to 1% mass percent concentration with the furantoin 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 furantoin, make it contain following component:
The fluorescent marker of the furantoin monoclonal antibody of FITC mark
The haptenic luminous marker of the furantoin of ABEI mark
The separation agent of the paramagnetism nano microsphere of pan coating goat-anti FITC monoclonal antibody
The furantoin standard solution (0ng/ml, 0.01ng/ml, 0.03ng/ml, 0.09ng/ml, 0.27ng/ml, 0.81ng/ml), the standard items dilution is pH7.4, contains 0.05% thimerosal antiseptic, the 0.1mol/LTRIS damping fluid.Said percentage composition is the quality percentage composition.
Furantoin quality-control product solution concentration is respectively 0.02ng/ml, 0.5ng/ml, and the quality-control product dilution contains 0.05% thimerosal antiseptic, the 0.1mol/LTRIS damping fluid for pH7.4.Said percentage composition is the quality percentage composition.
Concentrated washing lotion is pH7.2,0.4% Tween-20,0.3% sodium azide, 0.1mol/L PBS damping fluid.Said percentage composition is the quality percentage composition.
The detection of furantoin in embodiment three actual samples
1, feed sample pre-treatment
Take by weighing 1.0g ± 0.05g feed sample to the 50ml centrifuge tube, add 8ml ethyl acetate, add the 2ml acetonitrile again; Vibration 5min, static 10min, more than the room temperature 3000g, centrifugal 10min gets in the clean glass test tube of supernatant liquor 2ml to 10ml, flows down in 50～60 ℃ of water-bath nitrogen to dry up; Add the 1ml normal hexane,, add 1ml redissolution working fluid again, with vortex appearance whirling motion 30s with vortex appearance whirling motion 1min; More than the 3000g, the centrifugal 10min of room temperature (20-25 ℃); Remove upper organic phase, take off layer water 200 μ l and 200 μ l redissolution working fluid mixing; Getting 50 μ l is used for analyzing diluted sample multiple: 10.
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 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 furantoin and RLU proportion relation in the sample can be through the concentration of RLU combined standard curve method calculating furantoin.
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 furantoin standard items (0ng/ml, 0.01ng/ml, 0.03ng/ml, 0.09ng/ml, 0.27ng/ml 0.81ng/ml) carries out plotting curves.Instrument detects according to said method and obtains a RLU value and the concentration dependent calibration curve of furantoin, and in after this measuring, the furantoin concentration in each sample is compared with typical curve and drawn the residual content of the furantoin 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; Furantoin with 0.2ng/g, two concentration of 0.4ng/g adds recovery to the feed sample; 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
Can know from table, the average recovery rate scope that two concentration of furantoin are all added in the feed sample between 93.6-94.2%, 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 |
| Furantoin | 100％ |
| Furantoin | 15.3％ |
| Furaltadone | Less than 1% |
| Furazolidone | Less than 1% |
| AMOZ | Less than 0.1% |
| Furaxone metabolite | Less than 1% |
| Nitrofurazone (nitrofurazone) | Less than 1% |
| Nitrofurazone (nitrofurazone) metabolin | Less than 1% |
4, correlativity
X＝CI-2008，Y＝RIA
Y＝1.23X-2.57
R＝0.9815
X is CI-2008 system measurement result, and Y is that the result is measured in the aspect.
Claims (8)
1. magnetic granule chemoluminescence kit that detects furantoin, 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 furantoin haptens 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 furantoin 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 furantoin 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 furantoin and RLU proportion relation in the sample can be through the concentration of RLU set calibration curve method calculating furantoin.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103645310A (en) * | 2013-11-18 | 2014-03-19 | 洛阳莱普生信息科技有限公司 | Macrodantin chemiluminescence detection kit |
Citations (5)
| 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 |
| CN101029897A (en) * | 2007-02-09 | 2007-09-05 | 深圳市新产业生物医学工程有限公司 | Calcitonin reagent unit and its testing method |
| CN101393212A (en) * | 2008-11-04 | 2009-03-25 | 山东大学 | Chemiluminescence ELISA detection kit of furadantin |
| CN101551389A (en) * | 2009-01-14 | 2009-10-07 | 天津九鼎医学生物工程有限公司 | Magnetic particle chemiluminescence detection kit of free thyroxine and application thereof |
| CN101614742A (en) * | 2009-07-23 | 2009-12-30 | 清华大学 | The magnetic microparticle chemiluminescence immune assay determination kit of luteinizing principle |
-
2010
- 2010-12-16 CN CN2010105929251A patent/CN102539414A/en active Pending
Patent Citations (5)
| 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 |
| CN101029897A (en) * | 2007-02-09 | 2007-09-05 | 深圳市新产业生物医学工程有限公司 | Calcitonin reagent unit and its testing method |
| CN101393212A (en) * | 2008-11-04 | 2009-03-25 | 山东大学 | Chemiluminescence ELISA detection kit of furadantin |
| CN101551389A (en) * | 2009-01-14 | 2009-10-07 | 天津九鼎医学生物工程有限公司 | Magnetic particle chemiluminescence detection kit of free thyroxine and application thereof |
| CN101614742A (en) * | 2009-07-23 | 2009-12-30 | 清华大学 | The magnetic microparticle chemiluminescence immune assay determination kit of luteinizing principle |
Non-Patent Citations (3)
| Title |
|---|
| 《中国卫生检验杂志》 20100531 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 1234-1235 1-8 第20卷, 第5期 * |
| 李耀平等: "水产品中硝基呋喃代谢物残留快速检测新方法的研究", 《分析测试学报》 * |
| 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 (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103645310A (en) * | 2013-11-18 | 2014-03-19 | 洛阳莱普生信息科技有限公司 | Macrodantin chemiluminescence detection kit |
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Application publication date: 20120704 |