CN1445548A - Nano-colloidal gold marker immunization measurement method for testing carbofuran pesticide - Google Patents
Nano-colloidal gold marker immunization measurement method for testing carbofuran pesticide Download PDFInfo
- Publication number
- CN1445548A CN1445548A CN 03116692 CN03116692A CN1445548A CN 1445548 A CN1445548 A CN 1445548A CN 03116692 CN03116692 CN 03116692 CN 03116692 A CN03116692 A CN 03116692A CN 1445548 A CN1445548 A CN 1445548A
- Authority
- CN
- China
- Prior art keywords
- carbofuran
- antibody
- agricultural chemicals
- colloid gold
- gold label
- 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
Landscapes
- Peptides Or Proteins (AREA)
Abstract
A nano colloidal golf linked immunoassary for detecting carbofuran includes such steps as coupling the immunized semi-antigen of carbofuran with BSA, synthesizing artificial immunoantigen compound, immunizing animal to prepare specific high-valence BFNH antibody, separating and purifying, linking the nano-class colloidal golf to said specific antibody, solidifying said linked compound, carbofuran coupled by egg albumin and sheep's mouse IgG antibody on a carrier, and for semi-quantitative analysis. Its advantage is high sensitivity and speed.
Description
Technical field
What the present invention relates to is a kind of the immunochemistry detection technique to be applied to the detection of persticide residue, especially a kind of nano colloid gold label immunoassay method of carbofuran agricultural chemicals.Belong to the persticide residue detection range.
Background technology
Carbofuran (2,3-dihydro-2,2-dimethyl-7-benzofuryl methyl carbamate) has another name called carbofuran, be wide spectrum, efficient, interior absorption desinsection that the residue of drug effect phase is long, kill mite, nematicide, be widely used in the control of insect of crops such as paddy rice, corn, tobacco, tea, soybean, glue are white, leek.By the toxicological study of carbofuran being found carbofuran forms complex compound with cholinesterase in body, suppress its activity, acetylcholine can not be decomposed and put aside in vivo since the seventies, influence the impulse conduction between axoneure, have neurotoxicity.The residual term of validity is long in soil, drug effect is high for this medicament, can absorb conduction and be transported to each organ of plant by root, stem, leaf or seed, also can pass through runoff osmosis pollution underground water, health and environment are caused great harm, therefore, reinforcement is very necessary to the detection of carbofuran residues of pesticides.Up to now, the detection method of its residual quantity is mainly adopted physics ways such as liquid chromatography, gas chromatography and spectrum, but need expensive instrument and consuming time, be not suitable for the check and analysis of certain scaleization.By literature search, find: Development of Monoclonal Antibody-based Immunoassays to theN-Methylcarbamate Pesticide Carbofuran (foundation of the monoclonal antibody immunoassays of methyl carbamate agricultural chemicals carbofuran), see for details: Journal of Agricultural andFood Chemistry (agricultural and food chemistry magazine) 1999,47,2475-2485.Listed document has synthesized carbofuran artificial immunity antigen, and with the antigen immune mouse of synthetic, adopt hybridoma technology to prepare anti-carbofuran monoclonal antibody, set up the technology that indirect and direct enzyme linked immunosorbent assay (ELISA) detects the carbofuran agricultural chemicals on this basis.This technology is in former residues of pesticides conventional sense (gas chromatography, liquid chromatography instrument analytical approach) reforms on the basis of method, adopt enzyme linked immunosorbent assay (ELISA) technology (ELISA) to carry out Detecting Pesticide, make shorten detection time, cost reduces greatly, sensitivity improves, help at food, the fast detecting of carbofuran agricultural chemicals and be convenient to promote the use of widely in soil and the environment, it is numerous relatively tired that but its method remains in operation, need repeat repeatedly to wash, need measure by means of the microplate reader reading, can not realize real single stage method fast detecting.
Summary of the invention
The objective of the invention is to overcome deficiency of the prior art, a kind of nano colloid gold label immunoassay method of carbofuran agricultural chemicals is provided, make nanometer technology and immunological technique be applied to the Detecting Pesticide field, change the cumbersome detection problem of traditional residues of pesticides multistep, realized the method for single stage method fast detecting carbofuran residues of pesticides.
The present invention is achieved by the following technical solutions, the present invention with modified activity ester method with carbofuran immunity haptens (BFNH) and BSA coupling, synthetic artificial immunity antigenic compound, immune animal, the anti-BFNH antibody of preparation specific efficient valency, antibody is after separation and purification, with the specific antibody of nano level colloid gold label in anti-carbofuran, then with this mark bond, the carbofuran of coupling ovalbumin and sheep anti-mouse igg are solidificated in respectively on the carrier, suppress the immunochromatography principle according to competition, carrying out the semi-quantitative analysis of carbofuran agricultural chemicals measures, the detection architecture of being set up is 0.25mg/kg to the minimum detectable quantity of carbofuran agricultural chemicals, and finished in 10 minutes, method is stable, fast, accurately, be suitable for carrying out the fast detecting of single stage method residues of pesticides.Below technical scheme of the present invention made further specify, its concrete step is:
(1) preparation of artificial immunity compound
Adopt 2,3-dihydro-2,2-dimethyl-7-benzofuran phenol is a raw material, Synthetic 2 under the condition of logical phosgene, 3-dihydro-2,2-dimethyl-7-benzofuranyl chloro-carbonate, under cryogenic conditions, synthesize (BFNH) immune haptens with the 6-aminocaprolc acid reaction, adopt synthetic respectively immunizing antigen of modified activity ester method and envelope antigen then, respectively the 0.026g haptens is dissolved in the 1ml dry DMF, add 0.016g DCC and 0.009g NHs again, reaction is spent the night under the room temperature, the centrifuging and taking supernatant, dropwise add respectively during the BSA of 15mg/ml and OVA carbonic acid buffering tucks in, magnetic agitation 4 hours, distill water dialysis 2 times, normal saline dialysis 2 times.Packing is frozen in-20 ℃ of refrigerators.
(2) preparation of anti-carbofuran specific antibody
With synthetic BFNH-BSA immunity BALB/ small white mouse or experiment new zealand white rabbit, initial immunity is used the Fu Shi Freund's complete adjuvant emulsification artificial antigen of equivalent, then with freund 's incomplete adjuvant emulsification antigen, carry out fundamental immunity six times, adopt the ELISA method to detect serum antibody titer, treat that serum antibody reaches and carry out last booster immunization after necessarily tiring, take rabbit hearts serum.Or prepare monoclonal antibody with hybridoma technology.Separation and purification ,-20 ℃ frozen standby.
(3) preparation of collaurum
In the 100ml ultrapure water, add the gold chloride of 1ml 1% concentration, be heated to boiling, add 1% citric acid three sodium solution 1ml again, continued to boil 10 minutes, after the cooling, 4 ℃ of preservations are standby, and sampling sweep measuring maximum absorption band also carries out transmission electron microscope observing particle size.
(4) the anti-carbofuran specific antibody of colloid gold label
Regulating as above, the pH value of the colloidal gold solution (100ml) of preparation is 8.2, stir the anti-carbofuran specific antibody 400 μ g of adding down fast, making its final concentration is 4 μ g/ml, and room temperature reaction is after 15 minutes, adding polyglycol (PEG) to final concentration is 1%, continues to stir 15 minutes.With 8000r/min centrifugal 30 minutes, to inhale and go gained precipitation behind the supernatant to be the golden labelled antibody of purifying, this golden labelled antibody is suspended from again preserves in the liquid 4 ℃ of preservations.
(5) processing of the plain film of nitrocellulose filter and glass fibre
On nitrocellulose filter, carbofuran coupling ovalbumin bond is as p-wire with two parallel lines of little quantitation nozzle spraying, and sheep anti-mouse igg is as nature controlling line.On the plain film of glass fibre, standby behind the plain film of dry nitrocellulose filter and glass fibre with the anti-carbofuran antibody of quantitation nozzle spraying colloid gold label.
(6) preparation of immuno-chromatographic test paper strip
On the single face glue PVC of 60mm * 300mm plate, paste the upper glass cellulose membrane successively, be fixed with the plain film of glass fibre of the anti-carbofuran antibody of golden mark, parallel bag by the nitrocellulose filter of carbofuran-ovalbumin bond and anti-mouse IgG antibody, water adsorption glass cellulose membrane.The PVC plate that pastes vertically is cut into the test strips of 5mm * 60mm with cutting cutter
(7) sample test and determination methods as a result
Test strips is inserted in the testing sample, stopped to take out after 5 seconds and keep flat, or on sample pad, drip 3-4 drop of liquid sample, observations after 10 minutes with dropper.Sheep anti-mouse igg and carbofuran coupling OVA bond are sprayed on the nature controlling line (C) and the p-wire (T) of NC film respectively, containing the carbofuran testing sample moves to the other end according to chromatographic theory through the S end, and priority is crossed p-wire and nature controlling line, be solidificated in anti-carbofuran specific antibody of golden mark on the glass fibre membrane and the carbofuran in the sample and play specific reaction, and suppress it competitively and combine with carbofuran on the p-wire, therefore when the carbofuran content in the sample when a certain amount of, the anti-carbofuran specific antibody of gold mark combines back gold grain aggegation colour developing with the p-wire carbofuran, when the carbofuran content in the sample surpasses when a certain amount of, gold mark specific antibody is suppressed fully and does not develop the color, nature controlling line (C) is whether effectively the method for inspection itself is set, colour developing is effective, and not developing the color shows that method itself is invalid.
The present invention has substantive distinguishing features and marked improvement, and it is easy mainly to show as operation, does not need enzyme, does not need complicated numerous tired washing link; Sensitivity is stable, whether the exceeding standard of the agricultural chemicals to be measured in can the fast detecting sample, and be convenient to preserve; Detection speed is fast, and the entire reaction time finished within 10 minutes, can realize the needs of fast detecting; Nanometer technology and immunological technique are applied to the Detecting Pesticide field, have changed the cumbersome detection problem of traditional residues of pesticides multistep, realize the method for single stage method fast detecting carbofuran residues of pesticides.
Embodiment
Embodiment one:
Adopt hot method to synthesize immune haptens, get benzofuranol 100g and add the 250ml there-necked flask, add 19g catalyzer trimethyl benzyl ammonia chloride again, with being heated to 95 ℃ under the electronic stirring, feed phosgene with certain flow, precipitation occurred in 2 hours, structural characterization is carried out in sampling, gas spectrum, mass spectrophotometry.Taking by weighing the 4.6g6-aminocaproic acid then is dissolved in the NaOH solution of 6.4ml 4mol/L, add the 250ml flask, ice bath is cooled to 4 ℃, taking by weighing 7.18g benzofuranyl chloro-carbonate is dissolved in the 20ml diox, dividing 5 minor ticks to add in the there-necked flask in 10 minutes under stirring state reacted two hours, regulate pH value to 4.0, three extractions of ethyl acetate, separate out white precipitate, the washing oven dry.
Then with synthetic respectively immunizing antigen of modified activity ester method and envelope antigen, respectively the 0.026g haptens is dissolved in the 1ml dry DMF, add 0.016g DCC and 0.009g NHs again, reaction is spent the night under the room temperature, the centrifuging and taking supernatant dropwise adds during the BSA of 15mg/ml and OVA carbonic acid buffering tucks in magnetic agitation 4 hours respectively, distill water dialysis 2 times, normal saline dialysis 2 times; With synthetic BFNH-BSA immunity BALB/ small white mouse animal, initial immunity is used the Fu Shi Freund's complete adjuvant emulsification artificial antigen of equivalent, carry out abdominal part hypodermic, then with freund 's incomplete adjuvant emulsification antigen, carry out fundamental immunity six times, adopt the ELISA method to detect serum antibody titer, treat that serum antibody reaches and carry out last booster immunization after necessarily tiring, operation is got immune mouse spleen cell and the oncocyte hybridization of bone Sui, the HAT screening is also carried out cloning, filter out the cell line of the anti-carbofuran monoclonal antibody of energy stably excreting, cell line is injected mouse peritoneal, get ascites and prepare monoclonal antibody;
Then will be through 400 μ g of separation and purification, the pH that monoclonal antibody adds previously prepared 40nm size is in 8.2 the colloidal gold solution, room temperature reaction 15 minutes, adding poly-7 glycol (PEG) to final concentration is 1%, continues to stir 15 minutes.With 8000r/min centrifugal 30 minutes, inhale and go gained precipitation behind the supernatant to be the golden labelled antibody of purifying.This golden labelled antibody bond is sprayed on the plain film of glass fibre, carbofuran coupling-OVA bond is sprayed at the NC film, as p-wire, sheep anti-mouse igg is sprayed at the NC film, as nature controlling line, then with the absorption of sample pad, be coated with colloidal gold antibody glass fibre, the NC film that is sprayed with p-wire and nature controlling line and water adsorption glass fiber be pasted on a plastic bottom board (5mm * 6mm) successively.
During specimen, test strips is inserted in the testing sample, stop to take out after 5 seconds and keep flat, or on sample pad, drip 3-4 drop of liquid sample, observations after 10 minutes with dropper, if have only nature controlling line the aubergine band to occur, the result is positive, if the aubergine band all appears in nature controlling line and p-wire, the result is negative, if the aubergine band does not appear in nature controlling line, think that test result is invalid.
Embodiment two:
Adopt hot method to synthesize immune haptens, get benzofuranol 100g and add the 250ml there-necked flask, add 19g catalyzer trimethyl benzyl ammonia chloride again, with being heated to 95 ℃ under the electronic stirring, feed phosgene with certain flow, precipitation occurred in 2 hours, structural characterization is carried out in sampling, gas spectrum, mass spectrophotometry.Taking by weighing the 4.6g6-aminocaproic acid then is dissolved in the NaOH solution of 6.4ml 4mol/L, add the 250ml flask, ice bath is cooled to 4 ℃, taking by weighing 7.18g benzofuranyl chloro-carbonate is dissolved in the 20ml diox, dividing 5 minor ticks to add in the there-necked flask in 10 minutes under stirring state reacted two hours, regulate pH value to 4.0, three extractions of ethyl acetate, separate out white precipitate, the washing oven dry.
Then with synthetic respectively immunizing antigen of modified activity ester method and envelope antigen, respectively the 0.026g haptens is dissolved in the 1ml dry DMF, add 0.016g DCC and 0.009g NHs again, reaction is spent the night under the room temperature, the centrifuging and taking supernatant dropwise adds during the BSA of 15mg/ml and OVA carbonic acid buffering tucks in magnetic agitation 4 hours respectively, distill water dialysis 2 times, normal saline dialysis 2 times; With the synthetic athletic adult new zealand white rabbit of BFNH-BSA immunity, initial immunity is used the artificial antigen of 2.5ml Fu Shi Freund's complete adjuvant emulsification equivalent, carry out back intracutaneous multiple spot and huckle intramuscular injection, then with freund 's incomplete adjuvant emulsification antigen, carry out fundamental immunity six times, adopt the ELISA method to detect serum antibody titer, treat that serum antibody reaches and carry out last booster immunization after necessarily tiring, carried out the heart blood sampling every 7 days, the 25-30ml/ rabbit, separate purification serum ,-20 ℃ frozen standby.
Then will be through 400 μ g of separation and purification, the pH that monoclonal antibody adds previously prepared 40nm size is in 8.2 the colloidal gold solution, room temperature reaction 15 minutes, adding polyglycol (PEG) to final concentration is 1%, continues to stir 15 minutes.With 8000r/min centrifugal 30 minutes, inhale and go gained precipitation behind the supernatant to be the golden labelled antibody of purifying.This golden labelled antibody bond is sprayed on the plain film of glass fibre, carbofuran coupling-OVA bond is sprayed at the NC film, as p-wire, sheep anti-mouse igg is sprayed at the NC film, as nature controlling line, then with the absorption of sample pad, be coated with colloidal gold antibody glass fibre, the NC film that is sprayed with p-wire and nature controlling line and water adsorption glass fiber be pasted on a plastic bottom board (5mm * 6mm) successively.
During specimen, test strips is inserted in the testing sample, stop to take out after 5 seconds and keep flat, or on sample pad, drip 3-4 drop of liquid sample, observations after 10 minutes with dropper, if have only nature controlling line the aubergine band to occur, the result is positive, if the aubergine band all appears in nature controlling line and p-wire, the result is negative, if the aubergine band does not appear in nature controlling line, think that test result is invalid.
Claims (9)
1, a kind of nano colloid gold label immunoassay method of carbofuran agricultural chemicals, it is characterized in that with modified activity ester method carbofuran immunity haptens BFNH and BSA coupling, synthetic artificial immunity antigenic compound, immune animal, the anti-BFNH antibody of preparation specific efficient valency, antibody is after separation and purification, with the specific antibody of nano level colloid gold label in anti-carbofuran, then with this mark bond, the carbofuran of coupling ovalbumin and sheep anti-mouse igg are solidificated in respectively on the carrier, suppress the immunochromatography principle according to competition, carrying out the semi-quantitative analysis of carbofuran agricultural chemicals measures, the detection architecture of being set up is 0.25mg/kg to the minimum detectable quantity of carbofuran agricultural chemicals, and finishes fast detecting in 10 minutes.
2, the nano colloid gold label immunoassay method of carbofuran agricultural chemicals according to claim 1 is characterized in that, below technical scheme of the present invention is made further qualification, and its concrete step is:
(1) preparation of artificial immunity compound,
(2) preparation of anti-carbofuran specific antibody,
(3) preparation of collaurum,
(4) the anti-carbofuran specific antibody of colloid gold label,
(5) processing of the plain film of nitrocellulose filter and glass fibre,
(6) preparation of immuno-chromatographic test paper strip,
(7) sample test and determination methods as a result.
3, the nano colloid gold label immunoassay method of carbofuran agricultural chemicals according to claim 2, it is characterized in that, the concrete implication of related step (1) is: adopt 2,3-dihydro-2,2-dimethyl-7-benzofuran phenol is a raw material, Synthetic 2 under the condition of logical phosgene, 3-dihydro-2,2-dimethyl-7-benzofuranyl chloro-carbonate, under cryogenic conditions, synthesize BFNH immunity haptens with the 6-aminocaprolc acid reaction, adopt synthetic respectively immunizing antigen of modified activity ester method and envelope antigen then, respectively the 0.026g haptens is dissolved in the 1ml dry DMF, add 0.016g DCC and 0.009g NHs again, reaction is spent the night under the room temperature, the centrifuging and taking supernatant, dropwise add during the BSA of 15mg/ml and OVA carbonic acid buffering tucks in magnetic agitation 4 hours, distill water dialysis 2 times respectively, normal saline dialysis 2 times, packing is frozen in-20 ℃ of refrigerators.
4, the nano colloid gold label immunoassay method of carbofuran agricultural chemicals according to claim 2, it is characterized in that, the concrete implication of related step (2) is: with synthetic BFNH-BSA immunity BALB/ small white mouse or experiment new zealand white rabbit, initial immunity is used the Fu Shi Freund's complete adjuvant emulsification artificial antigen of equivalent, then with freund 's incomplete adjuvant emulsification antigen, carry out fundamental immunity six times, adopt the ELISA method to detect serum antibody titer, treat that serum antibody reaches and carry out last booster immunization after necessarily tiring, take rabbit hearts serum.Or prepare monoclonal antibody with hybridoma technology.Separation and purification ,-20 ℃ frozen standby.
5, the nano colloid gold label immunoassay method of carbofuran agricultural chemicals according to claim 2, it is characterized in that, the concrete implication of related step (3) is: the gold chloride that adds 1ml 1% concentration in the 100ml ultrapure water, be heated to boiling, add 1% citric acid three sodium solution 1ml again, continued to boil 10 minutes, after the cooling, 4 ℃ of preservations are standby, and sampling sweep measuring maximum absorption band also carries out transmission electron microscope observing particle size.
6, the nano colloid gold label immunoassay method of carbofuran agricultural chemicals according to claim 2, it is characterized in that, the concrete implication of related step (4) is: regulating as above, the pH value of the colloidal gold solution 100ml of preparation is 8.2, stir fast and add anti-carbofuran monoclonal antibody 400 μ g down, making its final concentration is 4 μ g/ml, behind the room temperature reaction 15 minutes, add polyglycol PEG to final concentration be 1%, continue to stir 15 minutes, with 8000r/min centrifugal 30 minutes, gained precipitated the golden labelled antibody that is purifying after supernatant was removed in suction, and this golden labelled antibody is suspended from again preserves in the liquid 4 ℃ of preservations.
7, the nano colloid gold label immunoassay method of carbofuran agricultural chemicals according to claim 2, it is characterized in that, the concrete implication of related step (5) is: spray two parallel lines on nitrocellulose filter with little quantitation nozzle, carbofuran coupling ovalbumin bond is as p-wire, and sheep anti-mouse igg is as nature controlling line.Spray the anti-carbofuran antibody of golden mark on the plain film of glass fibre with quantitation nozzle, standby behind the plain film of dry nitrocellulose filter and glass fibre.
8, the nano colloid gold label immunoassay method of carbofuran agricultural chemicals according to claim 2, it is characterized in that, the concrete implication of related step (6) is: on the single face glue PVC of 60mm * 300mm plate, paste the upper glass cellulose membrane successively, be fixed with the plain film of glass fibre of the anti-carbofuran antibody of golden mark, parallel bag by the nitrocellulose filter of carbofuran-ovalbumin and anti-mouse IgG antibody, water adsorption glass cellulose membrane, the PVC plate that pastes vertically is cut into the test strips of 5mm * 60mm with cutting cutter.
9, the nano colloid gold label immunoassay method of carbofuran agricultural chemicals according to claim 2, it is characterized in that, the concrete implication of related step (7) is: test strips is inserted in the testing sample, stop to take out after 5 seconds and keep flat, or on sample pad, drip 3-4 drop of liquid sample with dropper, observations after 10 minutes, sheep anti-mouse igg and carbofuran coupling OVA bond are sprayed on the nature controlling line (C) and the p-wire (T) of NC film respectively, containing the carbofuran testing sample moves to the other end according to chromatographic theory through the S end, and priority is crossed p-wire and nature controlling line, be solidificated in anti-carbofuran specific antibody of golden mark on the glass fibre membrane and the carbofuran in the sample and play specific reaction, and suppress it competitively and combine with carbofuran on the p-wire, therefore when the carbofuran content in the sample when a certain amount of, the anti-carbofuran specific antibody of gold mark combines back gold grain aggegation colour developing with the p-wire carbofuran, when the carbofuran content in the sample surpasses when a certain amount of, the gold labelled antibody is suppressed fully and does not develop the color, nature controlling line (C) is whether effectively the method for inspection itself is set, colour developing is effective, and not developing the color shows that method itself is invalid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03116692 CN1203317C (en) | 2003-04-29 | 2003-04-29 | Nano-colloidal gold marker immunization measurement method for testing carbofuran pesticide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03116692 CN1203317C (en) | 2003-04-29 | 2003-04-29 | Nano-colloidal gold marker immunization measurement method for testing carbofuran pesticide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1445548A true CN1445548A (en) | 2003-10-01 |
| CN1203317C CN1203317C (en) | 2005-05-25 |
Family
ID=27814918
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 03116692 Expired - Fee Related CN1203317C (en) | 2003-04-29 | 2003-04-29 | Nano-colloidal gold marker immunization measurement method for testing carbofuran pesticide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1203317C (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101806797A (en) * | 2010-03-17 | 2010-08-18 | 华中科技大学同济医学院附属同济医院 | Improved label immunoassay method |
| CN1866016B (en) * | 2006-06-08 | 2011-08-10 | 中国人民解放军军事医学科学院卫生学环境医学研究所 | Colloid gold immune test paper for detecting estradiol and preparation method thereof |
| CN103792350A (en) * | 2012-11-05 | 2014-05-14 | 江苏维赛科技生物发展有限公司 | Preparation method of fast detection reagent plate for detecting carbofuran |
| CN104849268A (en) * | 2015-05-21 | 2015-08-19 | 南京理工大学 | Aptamer wrapped gold nanoparticle probe-based reagent for colorimetric detection of omethoate and application of reagent |
| CN105699650A (en) * | 2016-02-16 | 2016-06-22 | 中国农业科学院农业质量标准与检测技术研究所 | Carbofuran bio-barcode immunoassay kit and application thereof |
| CN111141901A (en) * | 2020-02-14 | 2020-05-12 | 北京纳百生物科技有限公司 | Immune colloidal gold homogeneous phase mixed labeling method |
| CN113049812A (en) * | 2021-03-26 | 2021-06-29 | 信达安检测技术(天津)有限公司 | ELISA method for detecting carbofuran and kit thereof |
| CN113238043A (en) * | 2020-12-14 | 2021-08-10 | 黑龙江大学 | Preparation method and application of furadan test paper based on SERS immunochromatographic technique |
-
2003
- 2003-04-29 CN CN 03116692 patent/CN1203317C/en not_active Expired - Fee Related
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1866016B (en) * | 2006-06-08 | 2011-08-10 | 中国人民解放军军事医学科学院卫生学环境医学研究所 | Colloid gold immune test paper for detecting estradiol and preparation method thereof |
| CN101806797A (en) * | 2010-03-17 | 2010-08-18 | 华中科技大学同济医学院附属同济医院 | Improved label immunoassay method |
| CN101806797B (en) * | 2010-03-17 | 2014-02-19 | 华中科技大学同济医学院附属同济医院 | Improved label immunoassay method |
| CN103792350A (en) * | 2012-11-05 | 2014-05-14 | 江苏维赛科技生物发展有限公司 | Preparation method of fast detection reagent plate for detecting carbofuran |
| CN104849268B (en) * | 2015-05-21 | 2017-12-08 | 南京理工大学 | A kind of reagent of gold nanoparticle probe colorimetric detection flolimat based on aptamer parcel and its application |
| CN104849268A (en) * | 2015-05-21 | 2015-08-19 | 南京理工大学 | Aptamer wrapped gold nanoparticle probe-based reagent for colorimetric detection of omethoate and application of reagent |
| CN105699650A (en) * | 2016-02-16 | 2016-06-22 | 中国农业科学院农业质量标准与检测技术研究所 | Carbofuran bio-barcode immunoassay kit and application thereof |
| CN105699650B (en) * | 2016-02-16 | 2018-09-07 | 中国农业科学院农业质量标准与检测技术研究所 | Carbofuran bio-barcode immune analytic reagent kit and its application |
| CN111141901A (en) * | 2020-02-14 | 2020-05-12 | 北京纳百生物科技有限公司 | Immune colloidal gold homogeneous phase mixed labeling method |
| CN111141901B (en) * | 2020-02-14 | 2023-09-22 | 北京纳百生物科技有限公司 | Immune colloid Jin Junxiang mixed labeling method |
| CN113238043A (en) * | 2020-12-14 | 2021-08-10 | 黑龙江大学 | Preparation method and application of furadan test paper based on SERS immunochromatographic technique |
| CN113049812A (en) * | 2021-03-26 | 2021-06-29 | 信达安检测技术(天津)有限公司 | ELISA method for detecting carbofuran and kit thereof |
| CN113049812B (en) * | 2021-03-26 | 2024-03-19 | 信达安检测技术(天津)有限公司 | ELISA method for detecting carbofuran and kit thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1203317C (en) | 2005-05-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106918704B (en) | Synchronous detection aflatoxin and the time-resolved fluoroimmunoassay of carbaryl composite pollution chromatograph kit, preparation method and application | |
| US20180246086A1 (en) | Time-resolved fluorescent immunochromatographic assay rapid test kit for type i pyrethroid | |
| CN111187346B (en) | Colloidal gold test strip for detecting fipronil and metabolites thereof and preparation method thereof | |
| CN105950563B (en) | The monoclonal antibody and application of hybridoma cell strain 7E3 and its resistant to foot and mouth disease A type virus of secretion | |
| CN110441512B (en) | Colloidal gold immunochromatography detection device for ethyl maltol hapten and ethyl maltol | |
| CN1203317C (en) | Nano-colloidal gold marker immunization measurement method for testing carbofuran pesticide | |
| CN105481977A (en) | Glycocholic-acid immunodetection reagent based on anti-glycocholic acid specific antibody and preparation method thereof | |
| CN108196054A (en) | A kind of test strips for detecting glycyrrhizic acid and its preparation method and application | |
| CN108912090B (en) | Test strip for rapidly detecting total amount of alternariol and alternariol monomethyl ether | |
| CN108776219A (en) | A kind of immuno-chromatographic test paper strip of the thin Alternaria alternata ketone acid of quick detection | |
| CN108918851A (en) | A kind of preparation method of Lamotrigine colloidal gold strip | |
| CN109061146A (en) | A kind of test strips and its preparation method and application detecting Acetamiprid | |
| CN109187967A (en) | A kind of detection simultaneously distinguishes O-shaped, the duplex rapid detection card of A type foot and mouth disease virus and preparation method thereof | |
| CN106771273A (en) | One kind detection Formoterol colloidal gold immuno-chromatography test paper strip and preparation method and application | |
| CN103267842A (en) | Immune colloidal gold method for detecting diclofenac illegally added in Chinese patent medicament | |
| CN110927375A (en) | Fluorescent microsphere immunochromatography test strip for detecting olaquindox residue and application thereof | |
| CN113030463B (en) | Test strip for detecting impurities such as protein A in vaccine and application thereof | |
| CN110746286B (en) | Eugenol hapten, artificial antigen, preparation method and application thereof | |
| CN111154000A (en) | Anti-cimaterol monoclonal antibody and application thereof | |
| CN109580953A (en) | Mixed with spearmint colloidal gold detection device and preparation method and purposes in a kind of peppermint and its medicine materical crude slice | |
| CN111912988B (en) | Double-signal colloidal gold immunochromatographic test strip for detecting free gossypol as well as preparation method and application thereof | |
| CN116120242B (en) | Quick detection device for fenazaquin in tea, and preparation and application thereof | |
| CN109265364A (en) | A kind of preparation and application of pendimethalin haptens and antigen | |
| CN115215811B (en) | Prothioconazole hapten, antigen, antibody, detection device, preparation and application thereof | |
| CN113960307B (en) | Test strip for detecting isoprothiolane and preparation method 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 | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20050525 |