CN104101709A - Enzyme linked immunosorbent assay kit and detection method of rhodamine B - Google Patents
Enzyme linked immunosorbent assay kit and detection method of rhodamine B Download PDFInfo
- Publication number
- CN104101709A CN104101709A CN201310126445.XA CN201310126445A CN104101709A CN 104101709 A CN104101709 A CN 104101709A CN 201310126445 A CN201310126445 A CN 201310126445A CN 104101709 A CN104101709 A CN 104101709A
- Authority
- CN
- China
- Prior art keywords
- rhodamine
- kit
- sample
- concentration
- phosphate buffer
- 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.)
- Granted
Links
Classifications
-
- 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
-
- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
Abstract
The invention relates to an enzyme linked immunosorbent assay kit and a detection method of rhodamine B and belongs to the technical field of food safety detection. The kit is easy to operate and can sensitively, simply, conveniently, quickly and accurately detect content of the rhodamine B in a sample.
Description
Technical field
The present invention relates to food safety detection field.Particularly, the present invention relates to a kind of enzyme-linked immunologic detecting kit for detection of rhodamine B and detection method.
Background technology
Rhodamine B (Rhodamine B) claim again rose red b or basic rhodamine, is commonly called as pollen red, is the artificial synthetic dyestuffs of a kind of fresh pink.Find through zoopery, rhodamine B is carcinogen, does not therefore allow to add in food.Within 2008, the Ministry of Public Health issues " the non-edible material from soybeans of the illegal interpolation of possibility and the food additives kind list (first) of easy abuse in food ", clearly provides against and in food, adds rhodamine B class industrial dye.But still have a lot of illegal enterprises that rhodamine B is made an addition in flavouring and other food and is used as coloring agent.Therefore be badly in need of setting up a kind of detection method of rapid sensitive, for detection of illegal rhodamine B of adding in food.
At present, the detection of rhodamine B is mainly Laboratory Instruments analysis, and as high efficiency liquid phase-fluorescence detection and high efficiency liquid phase-mass spectrometry detection method, and immunoassay Related product is less on market.But lab analysis is higher with instrument and equipment and operation expense thereof, require also higher to operating personnel's specialty.And immunoassay is in instrumental analysis, pre-treating method is easy, detections is quick, can be used for Site Detection and be convenient to popularization.
Therefore, this area need to be prepared simply, and can detect sensitive, easy, quickly and accurately the enzyme linked immunological kit of the rhodamine B content in sample.
Summary of the invention
The object of this invention is to provide a kind of enzyme linked immunological kit and detection method, for detection of the rhodamine B of adding in food.
On the one hand, the invention provides the enzyme linked immunological kit for quantitatively detecting rhodamine B, described kit comprises: sample diluting liquid, it is the 0.01M phosphate buffer of the pH7.4 that contains 1% to 5% trehalose.
In one embodiment, described kit also comprises:
Be coated with the ELISA Plate of rhodamine B synthetic antigen, wherein the coated concentration of rhodamine B synthetic antigen is 0.5 to 1.5 μ g/mL;
Rhodamine B monoclonal antibody working fluid, it is the 0.01M phosphate buffer of the pH7.4 containing 0.5 to 1.0 μ g/mL rhodamine B monoclonal antibody;
The IgG antibody working fluid of horseradish peroxidase-labeled, it is the 0.01M phosphate buffer of the pH7.4 of the IgG antibody containing 0.5 to 1.0 μ g/mL horseradish peroxidase-labeled.
In one embodiment, described rhodamine B synthetic antigen is formulated in the 0.02M carbonate buffer solution of pH9.8.
In one embodiment, in described kit, the coated concentration of rhodamine B synthetic antigen is 1 μ g/mL.
In one embodiment, in described kit, rhodamine B monoclonal antibody working fluid is the 0.01M phosphate buffer containing the pH7.4 of 0.5 μ g/mL rhodamine B monoclonal antibody.
In one embodiment, in described kit, the IgG antibody working fluid of horseradish peroxidase-labeled is the 0.01M phosphate buffer containing the pH7.4 of the IgG antibody of 1 μ g/mL horseradish peroxidase-labeled.
In one embodiment, in described kit, sample diluting liquid is the 0.01M phosphate buffer of the pH7.4 that contains 1% trehalose.
In one embodiment, described kit also comprises:
Rhodamine B standard items;
Nitrite ion A, it is 5/10000ths to millesimal hydrogen peroxide or urea peroxide;
Nitrite ion B, it is 3/10000ths to 5/10000ths tetramethyl benzidine;
Stop buffer, it is 1.8 to 2.0mol/L sulfuric acid solution;
Cleansing solution, it is the phosphate buffer containing polysorbas20 0.1%-0.5%.
In one embodiment, described kit also comprises
Rhodamine B standard items;
Nitrite ion A, its hydrogen peroxide that is 5/10000ths or urea peroxide;
Nitrite ion B, its tetramethyl benzidine that is 3/10000ths;
Stop buffer, its sulfuric acid solution that is 2mol/L;
Cleansing solution, it is the phosphate buffer containing 0.1%-0.5% polysorbas20.
In the present invention, include but not limited to polystyrene, tygon or polypropylene for the preparation of the solid phase material of ELISA Plate, the form of carrier is concave hole type micro-reaction plate.
In one embodiment, described kit also comprises rhodamine B extract and operation instructions.
In one embodiment, described rhodamine B extract is methyl alcohol, methanol aqueous solution, acetonitrile or normal hexane.
On the other hand, the invention provides by enzyme linked immunological and test the method that detects the rhodamine B in sample, described method comprises use kit of the present invention.
In one embodiment, described method comprises:
With sample diluting liquid preparation rhodamine B standard items or testing sample.
In one embodiment, described method comprises:
In development step, every hole adds equal-volume nitrite ion A and B.
In one embodiment, described method comprises:
(1) be 0,0.1,0.3,0.9,2.7 and the rhodamine B standard items of 8.1ng/ml with sample diluting liquid compound concentration;
(2) in development step, every hole adds equal-volume nitrite ion A and B.
In one embodiment, described method also comprises the step of extracting the rhodamine B in testing sample.
In one embodiment, the step of extracting the rhodamine B in testing sample in described method comprises: (1) adds the extract of certain volume, and sample quality compares for 1:5 is to 1:10 with extracting liquid volume; (2) extraction time is 15 to 30 minutes; (3), after centrifugal or standing, get supernatant nitrogen and dry up; (4) add sample diluting liquid to redissolve, before and after nitrogen dries up, the volume ratio of solution is that 1:4 is to 1:5.
In one embodiment, the detection method of rhodamine B of the present invention comprises: (1) recovers room temperature by kit from preservation condition; (2) extract the rhodamine B (sample pre-treatments step) in testing sample; (3) preparation rhodamine B standard items working fluid; (4) in the hole of ELISA Plate, add successively IgG antibody working fluid and the antibody working fluid of standard items or sample, enzyme labeling; Hatch for (5) 25 DEG C; (6) washing; (7) in the hole of ELISA Plate, add nitrite ion; (8) 25 DEG C of colour developings; (9) add stop buffer; (10) in microplate reader in 450nm place reading; (11) calculate the content of rhodamine B in sample according to typical curve formula.
In one embodiment, sample-pretreating method comprises: (1) takes the sample of certain mass; (2) add the extract (including but not limited to, for example methyl alcohol, methanol aqueous solution, acetonitrile, normal hexane) of certain volume, sample quality with extracting liquid volume than arriving 1:10 for 1:5; (3) extraction time is 15 to 30 minutes; (4), after centrifugal or standing, get supernatant nitrogen and dry up; (5) add sample diluting liquid to redissolve, before and after nitrogen dries up, the volume ratio of solution is that 1:4 is to 1:5; (6) solution of getting after 50 μ L redissolve detects.
The detection principle of kit of the present invention is:
Rhodamine B synthetic antigen is coated on solid phase carrier; Add rhodamine B standard items or sample, and add IgG antibody and the rhodamine B antibody of enzyme labeling, rhodamine B fixing in the rhodamine B in sample and ELISA Plate competes the antibody in binding reagents, and the IgG antibody of enzyme labeling and rhodamine B antibody response; Remove and there is no the IgG of the rhodamine B of combination antibody and enzyme labeling antibody by washing; By enzymatic chromogenic reagent, after colour developing, stop the absorbance of working sample; The absorbance of sample and the concentration of rhodamine B are negative correlation, relatively can be calculated the concentration of rhodamine B in sample with typical curve.
Beneficial effect:
The present invention detects the kit of rhodamine B and mainly takes indirect competitive enzyme-linked immunosorbent determination method, qualitative or quantitatively detect the content of rhodamine B in sample; Testing process adopts a step to hatch method, has further shortened detection time, hatch indirect competitive with common two steps compared with, shortened to 45 minute by 1 hour 15 minutes detection time.The sample-pretreating method that this detection method provides is applicable to numerous food, and pretreatment process is simpler, can detect gross sample simultaneously.
Kit of the present invention uses rhodamine B monoclonal antibody, and main agents provides with working fluid form, for user's step that simplifies the operation, saves the running time.That the advantage of kit of the present invention is is highly sensitive, high specificity, accuracy are high, few to sample demand, to instrument and equipment require low, the reagent holding time is long etc., can in the food safety detection such as flavoring, play a significant role, have great social effect and wide market outlook.
Brief description of the drawings
Fig. 1 is the rhodamine B typical curve obtaining by kit of the present invention.
Fig. 2 is the rhodamine B typical curve obtaining by the large green health kit in river.
Fig. 3 is the typical curve while not containing trehalose in sample diluting liquid.
Embodiment
It is in order better further to understand the present invention that following embodiment is provided, and content of the present invention and protection domain is not formed to any restriction.
Main agents used source and composition in embodiment
Rhodamine B synthetic antigen and rhodamine B mouse resource monoclonal antibody are purchased from the excellent anti-how biological company limited in Guangzhou; The goat anti-mouse IgG antibody of horseradish peroxidase-labeled is purchased from the prompt Science and Technology Ltd. of Wuhan Amy; Rhodamine B standard items are purchased from Beijing North Na Chuanlian Bioteknologisk Institut.
Sample diluting liquid: 0.01M phosphate buffer, pH7.4, contains 1% trehalose
Carbonate buffer solution: 0.02M carbonate buffer solution, pH9.8
Cleansing solution: 0.01M phosphate buffer, pH7.4, containing 0.1%-0.5% polysorbas20
Confining liquid: 0.01M phosphate buffer, pH7.4, contains 1% BSA
Phosphate buffer: 0.01M phosphate buffer, pH7.4
Nitrite ion A: the hydrogen peroxide that concentration is 5/10000ths or urea peroxide solution
Nitrite ion B: the tetramethyl biphenyl amine aqueous solution that concentration is 3/10000ths
The sulfuric acid solution of stop buffer: 2mol/L
Embodiment 1: the foundation of indirect competitive ELISA method
1.1ELISA square formation is determined optimum response concentration
1.1.1 experimental technique
(1) coated: rhodamine B synthetic antigen (10mg/mL), coated with 5000,10000 or 20000 times of carbonate buffer solution dilutions, package amount is every hole 100 μ L, 4 DEG C are spent the night; With cleansing solution (containing 0.25% polysorbas20) washing three times, each 300 μ L/ holes, pat dry.(2) sealing: every hole adds 200 μ L confining liquids, 37 DEG C of incubations 3 hours; With cleansing solution washing three times, each 300 μ L/ holes, pat dry.(3) every hole adds 0.01M phosphate buffer (pH7.4) successively, the goat anti-mouse IgG antibody (diluting 1000 or 2000 times) of the horseradish peroxidase-labeled of diluting with 0.01M phosphate buffer (pH7.4) and the rhodamine B monoclonal antibody (diluting 5000,10000 or 20000 times) of diluting with 0.01M phosphate buffer (pH7.4), each 50 μ L; (4) hatch: incubated at room 30min; Afterwards, with cleansing solution washing five times.(5) colour developing: every hole adds nitrite ion A and the each 50 μ L of nitrite ion B, room temperature lucifuge effect 15min.(6) every hole adds 50 μ L stop buffers, detects the OD value at its 450nm place by microplate reader (Thermo microplate reader, model: MK3).(7) arrange simultaneously two parallel.The goat anti-mouse IgG antibody concentration of coated concentration, rhodamine B monoclonal anti bulk concentration and horseradish peroxidase-labeled while getting OD value 2.0 left and right is optium concentration.
1.1.2 experimental result:
Table 1: square formation method OD value
Data by table 1 can be determined, best antigen coated concentration is 10000 times (final concentration is 1 μ g/mL) of dilution, rhodamine B monoclonal antibody dilute concentration is 20000 times (final concentration is 0.5 μ g/mL) of dilution, and the goat anti-mouse IgG antibody dilution concentration of horseradish peroxidase-labeled is 1000 times (final concentration is 1 μ g/mL) of dilution.
1.2ELISA examination criteria curve IC50 value
1.2.1 except the antigen coated concentration of rhodamine B for dilution 10000 times,, rhodamine B monoclonal antibody dilute concentration for dilution 20000 times,, the goat anti-mouse IgG antibody dilution concentration of horseradish peroxidase-labeled is outside diluting 1000 times, the same 1.1.1 of ELISA method of operating.With sample diluting liquid compound concentration be 0,0.1,0.3,0.9,2.7 and the rhodamine B standard items of 8.1ng/mL.Every hole adds rhodamine B standard items successively, the each 50 μ L of the goat anti-mouse IgG antibody of horseradish peroxidase-labeled and rhodamine B monoclonal antibody; Incubated at room 30 minutes; Wash plate five times with cleansing solution; Develop the color 15 minutes.
1.2.2 testing result:
Table 2: typical curve OD value
| Rhodamine B concentration (ng/mL) | 0 | 0.1 | 0.3 | 0.9 | 2.7 | 8.1 |
| Average OD value | 1.855 | 1.734 | 1.713 | 1.099 | 0.423 | 0.146 |
Calculate IC50 in 1.2ng/mL left and right according to typical curve, show that the detection sensitivity of kit of the present invention, lower than 1.2ng/mL, can meet the testing requirement of rhodamine B.
The impact of trehalose on testing result in 1.3 sample diluting liquids
1.3.1 the same 1.2.1 of concentration of antigen concentration, rhodamine B monoclonal anti bulk concentration and enzyme labeling IgG antibody, sample diluting liquid is 0.01M phosphate buffer (pH7.4), does not contain trehalose, ELISA method of operating is with 1.1.1 and 1.2.1.
1.3.2 testing result
Table 3: typical curve OD value
| Rhodamine B concentration (ng/mL) | 0 | 0.1 | 0.3 | 0.9 | 2.7 | 8.1 |
| Average OD value | 1.755 | 1.740 | 1.697 | 1.282 | 0.689 | 0.241 |
Calculate IC50 in 1.9ng/mL left and right according to typical curve, according to 1.2.2, contain 1% trehalose in sample diluting liquid, IC50 is 1.2ng/mL left and right, shows that the trehalose in sample diluting liquid has improved the detection sensitivity of kit.
Embodiment 2: detect the establishment of the enzyme-linked immunologic detecting kit of rhodamine B
Set up the enzyme linked immunological kit that detects rhodamine B, make it comprise following component:
(1) be coated with the ELISA Plate of rhodamine B synthetic antigen: coating buffer is that ELISA Plate is polystyrene 96 orifice plates containing the 0.02M carbonate buffer solution of the pH9.8 of 1 μ g/mL antigen;
(2) high concentration rhodamine B standard items: concentration is 10 μ g/mL, need to specifications when use, and it is mixed with to concentration with sample diluting liquid is 0,0.1,0.3,0.9,2.7 and the standard items of 8.1ng/mL;
(3) concentration is the rhodamine B monoclonal antibody working fluid (preparing in the 0.01M of pH7.4 phosphate buffer) of 0.5 μ g/mL;
(4) concentration is the goat anti-mouse IgG antibody working fluid (preparing in the 0.01M of pH7.4 phosphate buffer) of the horseradish peroxidase-labeled of 1.0 μ g/mL;
(5) 10 × sample diluting liquids: 0.1M phosphate buffer, contains 10% trehalose;
(6) 20 × cleansing solutions: 0.2M phosphate buffer, contains 5% polysorbas20;
(7) nitrite ion A: the urea peroxide solution that concentration is 5/10000ths;
(8) nitrite ion B: the tetramethyl biphenyl amine aqueous solution that concentration is 3/10000ths;
(9) sulfuric acid solution of stop buffer: 2mol/L.
Embodiment 3: the detection of the rhodamine B of adding in sample
1 sample pre-treatments
(1) accurately take 1g sample (thick broad-bean sauce, thick chilli sauce, chilli powder etc.) in 50mL centrifuge tube; (2) add 10mL acetonitrile as extract, vibrate 15 minutes; (3) 4000rpm is centrifugal, gets supernatant nitrogen and dries up; (4) add sample diluting liquid (containing 1% trehalose) to redissolve; (5) solution of getting after 50 μ L redissolve detects.
2 use kits detect
(1) take out rhodamine B ELISA Plate, every hole adds standard solution (concentration be respectively 0,0.1,0.3,0.9,2.7 and 8.1ng/mL) or testing sample solution successively, the goat anti-mouse IgG antibody working fluid of horseradish peroxidase-labeled and rhodamine B monoclonal antibody working fluid, each 50 μ L; (2) hatch: room temperature, 30min; (3) washing: pour out liquid in hole, every hole adds 300 μ L cleansing solutions, vibration, pours out liquid in hole gently, repeats five times, pats dry with thieving paper.(4) colour developing: every hole adds nitrite ion A and the each 50 μ L of nitrite ion B, vibration mixes gently, room temperature lucifuge effect 15min.(5) every hole adds 50 μ L stop buffers, and vibration mixes gently, with microplate reader reading (OD
450nm).
3 interpretations of result
First calculate percentage absorptance: the mean value of the absorbance of standard items or sample (two multiple holes) is divided by the absorbance of negative standard items (concentration that is rhodamine B is 0ng/mL), then be multiplied by 100%, i.e. percentage absorbance (%)=B/B0 × 100%.Wherein: the mean light absorbency value of B-standard solution or sample solution; The concentration of B0-rhodamine B is the mean light absorbency value of the standard solution of 0ng/mL (ppb).
Then drawing standard curve result of calculation: taking rhodamine B standard items percentage absorptance as ordinate, the logarithm of rhodamine B standard items concentration (ng/mL) is horizontal ordinate, draws semilog canonical plotting (referring to Fig. 1).By in the percentage absorptance substitution typical curve of sample, read the corresponding concentration of sample from typical curve, be multiplied by its corresponding dilution factor and be rhodamine B actual amount in sample.Use kit specialty analysis software, can carry out accurate, the express-analysis of great amount of samples.
Embodiment 4: kit detectability experiment
The present embodiment detects the detectability of kit of the present invention.Concrete operations are:
(1) kit that prepared by the method from every crowd of embodiment 1 or 2, extracts a box; (2) detect sample diluting liquid, duplicate detection 20 times, (3) calculate the mean value (M) and the standard deviation (SD) that detect OD value for 20 times, and the corresponding absorbance of M-2SD: (4) bring corresponding M-2SD absorbance into typical curve, the concentration calculating is kit detectability, and result is 0.01ng/mL.
Embodiment 5: kit Precision Experiment
The present embodiment is that kit detects repeatable experiment.Concrete operations are:
(1) kit that prepared by the method from every crowd of embodiment 1 or 2, extracts a box; (2) detectable concentration is 2 and the detected value of the sample solution (pre-treating method is with embodiment 3) of 4ng/mL, each concentration duplicate detection 10 times; (3) calculate the coefficient of variation.
Result shows that the coefficient of variation is between 6% to 10%, has met the coefficient of variation and has been not more than 10% regulation, illustrates that the precision of kit of the present invention has reached requirement.
Embodiment 6: kit recovery experiment
In actual sample, add rhodamine B standard items, add 3 concentration, each concentration do two parallel, calculate recovery rate.Result shows that the interpolation recovery in thick broad-bean sauce is 75% to 115%.
Embodiment 7: kit storage life experiment
Kit preservation condition is 2 to 8 DEG C, through the mensuration of 6 months, the maximum absorbance value of kit (standard items that concentration is zero), IC50(50% inhibition concentration) with the interpolation recovery of rhodamine B all within normal range.Consider in transport and use procedure, have improper preservation condition and occur, carried out accelerated stability experiment, kit is preserved to the Ninth Heaven at 37 DEG C, detect rear discovery, kit indices still meets the requirements.Can show that from above result kit can be 2 to 8 DEG C of preservations 12 months.
Embodiment 8: kit contrast experiment
The present embodiment is the quality of the detection effect of kit more of the present invention and outsourcing kit, and concrete operations are:
(1) kit in two sources of use detects the concentration of rhodamine B in chilli powder, one of them kit of preparing for the present invention (referring to embodiment 2); Another is the kit that is purchased from river large green health biotechnology research company limited.(2) detection method of kit of the present invention and pre-treating method are respectively referring to embodiment 1 and embodiment 3.(3) detection of the large green health kit in river is carried out according to this kit instructions.(4) result shows that the typical curve linearly dependent coefficient of kit of the present invention is better than the large green health kit in river, and accuracy in detection is also better than the kit of the large green health in river.
Table 4: kit of the present invention detects rhodamine B concentration in chilli powder
Table 5: the large green health kit in river detects rhodamine B concentration in chilli powder
Claims (10)
1. for quantitatively detecting the enzyme linked immunological kit of rhodamine B, described kit comprises:
Sample diluting liquid, it is the 0.01M phosphate buffer of the pH7.4 that contains 1% to 5% trehalose.
2. kit as claimed in claim 1, it also comprises:
Be coated with the ELISA Plate of rhodamine B synthetic antigen, wherein the coated concentration of rhodamine B synthetic antigen is 0.5 to 1.5 μ g/mL;
Rhodamine B monoclonal antibody working fluid, it is for containing the 0.01M phosphate buffer of the pH7.4 that is 0.5 to 1.0 μ g/mL rhodamine B monoclonal antibody;
The IgG antibody working fluid of horseradish peroxidase-labeled, it is the 0.01M phosphate buffer of the pH7.4 of the IgG antibody containing 0.5 to 1.0 μ g/mL horseradish peroxidase-labeled.
3. kit as claimed in claim 1 or 2, it also comprises
Rhodamine B standard items;
Nitrite ion A, it is 5/10000ths to millesimal hydrogen peroxide or urea peroxide;
Nitrite ion B, it is 3/10000ths to 5/10000ths tetramethyl benzidine;
Stop buffer, it is 1.8 to 2.0mol/L sulfuric acid solution;
Cleansing solution, it is the phosphate buffer containing 0.1%-0.5% polysorbas20.
4. the kit as described in aforementioned any one claim, is wherein polystyrene, tygon or polypropylene for the preparation of the solid phase material of described ELISA Plate, and the form of carrier is concave hole type micro-reaction plate.
5. the kit as described in aforementioned any one claim, it also comprises rhodamine B extract and operation instructions.
6. kit as claimed in claim 5, wherein said rhodamine B extract is methyl alcohol, methanol aqueous solution, acetonitrile or normal hexane.
7. test by enzyme linked immunological the method that detects the rhodamine B in sample, described method comprises that right to use requires the kit described in any one in 1-6.
8. method as claimed in claim 7, it comprises
With sample diluting liquid preparation rhodamine B standard items or testing sample.
9. method as claimed in claim 7 or 8, it also comprises
In development step, every hole adds equal-volume nitrite ion A and B.
10. the method as described in claim 7-9 any one, it also comprises the step of extracting the rhodamine B in testing sample.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310126445.XA CN104101709B (en) | 2013-04-12 | 2013-04-12 | A kind of enzyme linked immunological kit detecting rhodamine B and detection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310126445.XA CN104101709B (en) | 2013-04-12 | 2013-04-12 | A kind of enzyme linked immunological kit detecting rhodamine B and detection method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104101709A true CN104101709A (en) | 2014-10-15 |
| CN104101709B CN104101709B (en) | 2016-08-31 |
Family
ID=51670029
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310126445.XA Active CN104101709B (en) | 2013-04-12 | 2013-04-12 | A kind of enzyme linked immunological kit detecting rhodamine B and detection method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104101709B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6485726B1 (en) * | 1995-01-17 | 2002-11-26 | The Brigham And Women's Hospital, Inc. | Receptor specific transepithelial transport of therapeutics |
| CN1425918A (en) * | 2003-01-14 | 2003-06-25 | 江苏省血吸虫病防治研究所 | Quick test paper strip diagnostic reagent kits for bilharziasis and its preparing method and use |
| CN101871936A (en) * | 2010-06-03 | 2010-10-27 | 江南大学 | Rhodamine B ELISA detection method |
| CN102297966A (en) * | 2011-05-27 | 2011-12-28 | 天津农学院 | Progesterone semi-quantitative determining colloidal gold test paper and preparation method thereof |
| CN102662062A (en) * | 2012-04-17 | 2012-09-12 | 中国农业科学院兰州兽医研究所 | Monoclonal antibody blocking enzyme-linked immunosorbent assay (ELISA) kit and method for detecting nonstructural protein (NSP) antibody of foot-and-mouth disease virus (FMDV) |
| CN102830229A (en) * | 2012-08-27 | 2012-12-19 | 北京新兴四寰生物技术有限公司 | Detection reagent of treponema pallidum IgM (Immunoglobulin M) antigen colloidal gold method and preparation method thereof |
-
2013
- 2013-04-12 CN CN201310126445.XA patent/CN104101709B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6485726B1 (en) * | 1995-01-17 | 2002-11-26 | The Brigham And Women's Hospital, Inc. | Receptor specific transepithelial transport of therapeutics |
| CN1425918A (en) * | 2003-01-14 | 2003-06-25 | 江苏省血吸虫病防治研究所 | Quick test paper strip diagnostic reagent kits for bilharziasis and its preparing method and use |
| CN101871936A (en) * | 2010-06-03 | 2010-10-27 | 江南大学 | Rhodamine B ELISA detection method |
| CN102297966A (en) * | 2011-05-27 | 2011-12-28 | 天津农学院 | Progesterone semi-quantitative determining colloidal gold test paper and preparation method thereof |
| CN102662062A (en) * | 2012-04-17 | 2012-09-12 | 中国农业科学院兰州兽医研究所 | Monoclonal antibody blocking enzyme-linked immunosorbent assay (ELISA) kit and method for detecting nonstructural protein (NSP) antibody of foot-and-mouth disease virus (FMDV) |
| CN102830229A (en) * | 2012-08-27 | 2012-12-19 | 北京新兴四寰生物技术有限公司 | Detection reagent of treponema pallidum IgM (Immunoglobulin M) antigen colloidal gold method and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 宋姗姗: "食品中罗丹明B的残留免疫检测方法的研究", 《中国优秀硕士学位论文全文数据库工程科技Ⅰ辑》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104101709B (en) | 2016-08-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109061204A (en) | A kind of kit of fluorescence immunoassay detection hair trace drugs | |
| CN103018458A (en) | Ultra-sensitive aflatoxin B1 enzyme-linked immunosorbent assay kit | |
| CN104076154A (en) | Enzyme linked immunosorbent assay kit detecting folic acid and application thereof | |
| Korpe et al. | Evaluation of a rapid point-of-care fecal antigen detection test for Entamoeba histolytica | |
| CN108089007A (en) | A kind of kit and preparation method for quantitatively detecting c reactive protein | |
| CN104897864A (en) | ELISA (enzyme linked immunosorbent assay) kit for detecting amantadine and application of ELISA kit | |
| CN209803153U (en) | Four-in-one detection kit for fluorescence-immune trace hair drugs | |
| CN105319368A (en) | Enzyme linked immunosorbent assay kit used for detecting zearalenone, and detection method thereof | |
| CN103513036B (en) | The CLEIA detection kit of salmonella and detection method | |
| CN112649610A (en) | Detection kit and detection method for methamphetamine drugs in hair | |
| CN103575885A (en) | Enzyme linked immunoassay kit for detecting T-2 toxin, and application thereof | |
| CN104558084A (en) | Method for extracting steroid hormone from excrement of forest musk deer | |
| CN104101709A (en) | Enzyme linked immunosorbent assay kit and detection method of rhodamine B | |
| Zhang et al. | Rapid monitoring of dicyclohexyl phthalate in foods using the direct competitive ELISA | |
| CN103645320B (en) | A kind of AMOZ chemiluminescent enzyme-linked immunosorbent immunity quick detection kit | |
| CN103226146B (en) | The detection method of the three-in-one euzymelinked immunosorbent assay (ELISA) of Clenbuterol, Ractopamine, salbutamol and dedicated kit and kit | |
| Premjeet et al. | Enzyme-Linked Immuno-Sorbent Assay (ELISA), basics and it’s application: A comprehensive review | |
| CN104142406A (en) | Stable troponin I detection kit | |
| CN202939176U (en) | ELISA (Enzyme Linked Immunosorbent Assay) test kit of walnut protein | |
| CN110618274B (en) | Chimeric ELISA kit for simultaneously and quantitatively detecting total amount of aflatoxin and zearalenone in edible oil | |
| CN202903791U (en) | T-2 toxin ELISA (enzyme-linked immunoabsorbent assay) detection reagent kit | |
| CN203465267U (en) | Triclabendazole enzyme-linked immunosorbent assay kit | |
| CN201984069U (en) | Melamine luminous ELIsA (enzyme linked immunosorbent assay) kit | |
| CN104569398A (en) | Enzyme-linked immunoassay kit for detecting ethoxyquinoline and application thereof | |
| CN206002549U (en) | A kind of human neutrophil apolipoprotein heterodimer proportioning device based on Enzyme-linked Immunosorbent Assay technology |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |