CN108469517A - Time-resolved fluorescent test strip for detecting trichlorfon and application thereof - Google Patents
Time-resolved fluorescent test strip for detecting trichlorfon and application thereof Download PDFInfo
- Publication number
- CN108469517A CN108469517A CN201810217658.6A CN201810217658A CN108469517A CN 108469517 A CN108469517 A CN 108469517A CN 201810217658 A CN201810217658 A CN 201810217658A CN 108469517 A CN108469517 A CN 108469517A
- Authority
- CN
- China
- Prior art keywords
- metrifonate
- detection
- absorption pad
- time
- sample
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- NFACJZMKEDPNKN-UHFFFAOYSA-N trichlorfon Chemical compound COP(=O)(OC)C(O)C(Cl)(Cl)Cl NFACJZMKEDPNKN-UHFFFAOYSA-N 0.000 title claims abstract description 90
- 229960001952 metrifonate Drugs 0.000 title claims abstract description 88
- 238000012360 testing method Methods 0.000 title claims abstract description 45
- 238000001514 detection method Methods 0.000 claims abstract description 40
- 238000010521 absorption reaction Methods 0.000 claims abstract description 26
- 238000006243 chemical reaction Methods 0.000 claims abstract description 19
- 239000000427 antigen Substances 0.000 claims abstract description 18
- 102000036639 antigens Human genes 0.000 claims abstract description 18
- 108091007433 antigens Proteins 0.000 claims abstract description 18
- 239000004005 microsphere Substances 0.000 claims abstract description 15
- 239000011248 coating agent Substances 0.000 claims abstract description 13
- 238000000576 coating method Methods 0.000 claims abstract description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 12
- 238000003908 quality control method Methods 0.000 claims abstract description 10
- 235000010149 Brassica rapa subsp chinensis Nutrition 0.000 claims abstract description 9
- 235000000536 Brassica rapa subsp pekinensis Nutrition 0.000 claims abstract description 9
- 241000499436 Brassica rapa subsp. pekinensis Species 0.000 claims abstract description 9
- 238000002360 preparation method Methods 0.000 claims description 13
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 12
- 102000004169 proteins and genes Human genes 0.000 claims description 12
- 108090000623 proteins and genes Proteins 0.000 claims description 12
- 241000207199 Citrus Species 0.000 claims description 9
- 235000020971 citrus fruits Nutrition 0.000 claims description 9
- 240000007124 Brassica oleracea Species 0.000 claims description 8
- 235000003899 Brassica oleracea var acephala Nutrition 0.000 claims description 8
- 235000011301 Brassica oleracea var capitata Nutrition 0.000 claims description 8
- 235000001169 Brassica oleracea var oleracea Nutrition 0.000 claims description 8
- 239000003550 marker Substances 0.000 claims description 6
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- -1 EDC hydrochloric acid Salt Chemical class 0.000 claims description 3
- 229960000549 4-dimethylaminophenol Drugs 0.000 claims description 2
- GOUHYARYYWKXHS-UHFFFAOYSA-N 4-formylbenzoic acid Chemical class OC(=O)C1=CC=C(C=O)C=C1 GOUHYARYYWKXHS-UHFFFAOYSA-N 0.000 claims description 2
- 230000006837 decompression Effects 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- 238000010792 warming Methods 0.000 claims description 2
- 238000004440 column chromatography Methods 0.000 claims 1
- 240000007594 Oryza sativa Species 0.000 abstract description 3
- 235000007164 Oryza sativa Nutrition 0.000 abstract description 3
- 235000009566 rice Nutrition 0.000 abstract description 3
- 238000012216 screening Methods 0.000 abstract description 3
- 230000035945 sensitivity Effects 0.000 abstract description 3
- 239000012528 membrane Substances 0.000 abstract 4
- 244000178937 Brassica oleracea var. capitata Species 0.000 abstract 1
- 244000141359 Malus pumila Species 0.000 abstract 1
- 244000269722 Thea sinensis Species 0.000 abstract 1
- 235000021016 apples Nutrition 0.000 abstract 1
- 238000012544 monitoring process Methods 0.000 abstract 1
- 235000013616 tea Nutrition 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical class OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 11
- 239000000020 Nitrocellulose Substances 0.000 description 5
- 239000006143 cell culture medium Substances 0.000 description 5
- 210000004408 hybridoma Anatomy 0.000 description 5
- 239000001257 hydrogen Substances 0.000 description 5
- 229910052739 hydrogen Inorganic materials 0.000 description 5
- 229920001220 nitrocellulos Polymers 0.000 description 5
- 239000007921 spray Substances 0.000 description 5
- 238000002965 ELISA Methods 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 238000001035 drying Methods 0.000 description 4
- 235000013399 edible fruits Nutrition 0.000 description 4
- 238000011587 new zealand white rabbit Methods 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 235000013311 vegetables Nutrition 0.000 description 4
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 3
- 241000283973 Oryctolagus cuniculus Species 0.000 description 3
- 241001494479 Pecora Species 0.000 description 3
- 239000004793 Polystyrene Substances 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000003317 immunochromatography Methods 0.000 description 3
- 239000004033 plastic Substances 0.000 description 3
- 229920002223 polystyrene Polymers 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 102000014914 Carrier Proteins Human genes 0.000 description 2
- 108010078791 Carrier Proteins Proteins 0.000 description 2
- 229930182566 Gentamicin Natural products 0.000 description 2
- 231100000703 Maximum Residue Limit Toxicity 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 206010035226 Plasma cell myeloma Diseases 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 241000607479 Yersinia pestis Species 0.000 description 2
- 125000003172 aldehyde group Chemical group 0.000 description 2
- 244000309466 calf Species 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 201000000050 myeloid neoplasm Diseases 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- 210000004988 splenocyte Anatomy 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 235000017060 Arachis glabrata Nutrition 0.000 description 1
- 244000105624 Arachis hypogaea Species 0.000 description 1
- 235000010777 Arachis hypogaea Nutrition 0.000 description 1
- 235000018262 Arachis monticola Nutrition 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 244000241257 Cucumis melo Species 0.000 description 1
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 description 1
- 229910052693 Europium Inorganic materials 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 108010044091 Globulins Proteins 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108010058846 Ovalbumin Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- YNPNZTXNASCQKK-UHFFFAOYSA-N Phenanthrene Natural products C1=CC=C2C3=CC=CC=C3C=CC2=C1 YNPNZTXNASCQKK-UHFFFAOYSA-N 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- 108010090804 Streptavidin Proteins 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 241000254234 Xyeloidea Species 0.000 description 1
- DGEZNRSVGBDHLK-UHFFFAOYSA-N [1,10]phenanthroline Chemical compound C1=CN=C2C3=NC=CC=C3C=CC2=C1 DGEZNRSVGBDHLK-UHFFFAOYSA-N 0.000 description 1
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical class N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000005875 antibody response Effects 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 108091006004 biotinylated proteins Proteins 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 235000021329 brown rice Nutrition 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 229920002301 cellulose acetate Polymers 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000009137 competitive binding Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- OGPBJKLSAFTDLK-UHFFFAOYSA-N europium atom Chemical compound [Eu] OGPBJKLSAFTDLK-UHFFFAOYSA-N 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 208000010824 fish disease Diseases 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- CEAZRRDELHUEMR-UHFFFAOYSA-N gentamicin Chemical class O1C(C(C)NC)CCC(N)C1OC1C(O)C(OC2C(C(NC)C(C)(O)CO2)O)C(N)CC1N CEAZRRDELHUEMR-UHFFFAOYSA-N 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 108060003552 hemocyanin Proteins 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000037189 immune system physiology Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- VTACLVUOTMPORB-UHFFFAOYSA-N n,n-bis(trimethylsilyl)acetamide Chemical class CC(=O)N([Si](C)(C)C)[Si](C)(C)C VTACLVUOTMPORB-UHFFFAOYSA-N 0.000 description 1
- 239000003986 organophosphate insecticide Substances 0.000 description 1
- 150000002903 organophosphorus compounds Chemical class 0.000 description 1
- 229940092253 ovalbumin Drugs 0.000 description 1
- 235000020232 peanut Nutrition 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 231100000175 potential carcinogenicity Toxicity 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000012372 quality testing Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 150000003625 trehaloses Chemical class 0.000 description 1
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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a time-resolved fluorescence test strip for detecting trichlorfon and application thereof. The test strip comprises a sample absorption pad, a reaction membrane and a water absorption pad, wherein the reaction membrane is positioned in the middle, and the sample absorption pad and the water absorption pad are respectively lapped at the left end and the right end of the reaction membrane; a sample adding area and a microsphere area are arranged on the sample absorption pad, and the microsphere area is loaded with trichlorfon time-resolved fluorescent microspheres; the reaction membrane is provided with a detection line (T line) and a quality control line (C line), the T line is connected with a coated dipterex antigen (coating antigen), and the C line is coated with an anti-rabbit antibody. The invention also provides a method for detecting trichlorfon residues in samples such as apples, rice, tea leaves, oranges, common head cabbages, Chinese cabbages and the like by applying the trichlorfon test strip. The test strip provided by the invention has the characteristics of simple operation, high sensitivity, quantitative detection, rapid detection, low cost and the like, and is suitable for screening and on-site monitoring of a large number of samples.
Description
Technical field
The present invention relates to a kind of time-resolved fluorescence test strips of detection metrifonate and its applications, belong to time-resolved fluorescence
Immunoassay (TRFIA) technical field, for metrifonate in the samples such as apple, paddy, tealeaves, citrus, cabbage, Chinese cabbage
Content detection.
Background technology
Metrifonate scientific name 0,0- dimethyl-(2,2,2- tri- chloro- 1- hydroxyethyls) phosphate, is that one kind being widely used in field
Between crops organophosphorus insecticide, have many advantages, such as low toxicity, efficiently, good water solubility, agriculturally can be used for green vegetables worm, trunk
A variety of pest controls such as worm, fruit tree sawfly can be used as a kind of good multiple-effect pest repellant in animal husbandry, can be used in aquaculture
In the prevention of fish disease.But metrifonate is easy to enter human body through respiratory tract and skin, and in recent years, potential carcinogenicity and cause
Mutability causes the great attention of people.Therefore, correlation quality testing department in China's has carried out limitation regulation to the substance.GB
2763-2016《National food safety standard Pesticide maximum residue limit》Define the maximum residue limit of metrifonate：
Paddy, brown rice, wheat, shelled peanut and soybean are 0.1mg/kg, and Chinese cabbage, cabbage are 0.1mg/kg, water in vegetables
Citrus, pomaceous fruit, kernel approaches, melon and fruit class etc. are 0.2mg/kg, tealeaves 2mg/kg in fruit.
Currently, the method for detection metrifonate has gas chromatography, high performance liquid chromatography etc..Instrument detection method it is accurate
Degree is higher, but expensive equipment, needs technical professional, detecting step complicated, it is difficult to which batch detection sample is not suitable for
Laboratories batch quickly detects sample.And immunological detection method is easy to operate, quick, sensitive, can detect majority simultaneously
Sample is ideal quick screening means.
Invention content
The object of the present invention is to provide a kind of high sensitivity, the metrifonate times that easy to operate, at low cost, detection time is short
Resolved fluorometric test strips.
A kind of metrifonate time-resolved fluorescence test strips provided by the present invention, which includes sample absorption pad, anti-
Answer film and water absorption pad three parts.Reaction film is located at centre, and sample absorption pad is overlapped in reaction film or so two respectively with water absorption pad
End.Wherein, sample absorption pad is equipped with sample application zone and microballoon area, and microballoon area is loaded with time-resolved fluorescence microballoon；On reaction film
Equipped with detection line (T lines) and nature controlling line (C lines), T lines connect coating metrifonate antigen, and C lines are coated with anti-rabbit antibody.
The metrifonate time-resolved fluorescence microballoon, including detection microballoon and Quality Control microballoon, detection microballoon are pan coating
It is the fluorescent microsphere that pan coating has rabbit-anti labelled protein to have the fluorescent microsphere of metrifonate monoclonal antibody, Quality Control microballoon.
Bright-coloured series elements compound is filled in the fluorescent microsphere；Preferably, which is europium chelating object；
Optimal, which can be Eu (TTA) 3/TOPO or Eu (TTA) 3/Phen.
The albumen can be cow's serum Y- globulin (BGG) or balf serum albumin (BSA).
The time-resolved fluorescence microspherulite diameter ranging from 100-1000nm.
On the nitrocellulose filter, coated antibody is metrifonate antigen on T lines, coated anti-rabbit antibody on C lines.
The metrifonate time-resolved fluoroimmunoassay quantitative testing test paper bottom is equipped with plastic bottom board.
The T lines, which connect coating metrifonate antigen, to be obtained with carrier protein couplet by metrifonate haptens, the carrier
Albumen can be bovine serum albumin(BSA), ovalbumin, hemocyanin, thyroprotein, human serum albumins.
The metrifonate monoclonal antibody is to prepare to obtain using metrifonate hapten-carrier protein conjugate as immunogene
, it is to be secreted to obtain by metrifonate monoclonal antibody hybridoma cell strain；The anti-rabbit antibody is to obtain rabbit source antibody mediated immunity sheep
It arrives.
The sample absorption pad is Fusion5 films or the identical film of other functions；The water absorption pad is water absorption pad；It is described anti-
It can be nitrocellulose filter or cellulose acetate film to answer film.
It is a further object to provide a kind of methods preparing above-mentioned test strips comprising step：
(1) prepared by Quality Control microballoon：
1. using biotinylated protein；
2. being coated with aldehyde group modified fluorescent microsphere using above-mentioned labelled protein；
(2) prepared by haptens：Metrifonate haptens product is obtained by series of chemical；
(3) by metrifonate haptens and carrier protein couplet, metrifonate hapten-carrier protein conjugate is obtained；
(4) use metrifonate hapten-carrier protein conjugate that new zealand white rabbit is immunized, by new zealand white rabbit splenocyte
With myeloma cell by merging, screening, metrifonate monoclonal hybridoma strain is obtained；
(5) new zealand white rabbit IgG immune health goats are extracted, anti-rabbit antibody is obtained；
(6) prepared by detection microballoon：Aldehyde group modified fluorescent microsphere is coated with using the monoclonal antibody of metrifonate；
(7) blank kilocalorie is pasted：Nitrocellulose filter is pasted onto among plastic bottom board, Fusion5 films and water absorption pad point
It is not overlapped in nitrocellulose filter left and right ends；
(8) film is sprayed：By the fluorescent microsphere of rabbit-anti labelled protein and the fluorescent microsphere of metrifonate monoclonal antibody using release
Buffer solution mixed diluting is sprayed onto the microballoon area of Fusion5 films to a certain concentration；After metrifonate antigen and the dilution of anti-rabbit antibody, point
It is not sprayed onto the T lines and C line positions of nitrocellulose filter；
(9) dry and slitting：By the above-mentioned kilocalorie drying for having sprayed reagent, slitting.
The release buffer solution used in spray film step contains 10%-20% sucrose, 3%-10% trehaloses, 0.5%-1%
The bis- trimethylsilyl acetamides (BSA) of N, 0-, 0.2%-0.5% gentamicins.
The final concentration of 0.5mg/mL-2mg/mL of microballoon, Quality Control microballoon final concentration 0.05mg/ are detected after spraying the dilution of film step
mL-0.2mg/mL；T line antigen final concentration 0.5mg/mL-2mg/mL, C line antigen final concentrations 0.5mg/mL-2mg/mL；C, T line spray
Film liquid amount is 0.5 μ L/cm-1.5 μ L/cm, and it is 2 μ L/cm-8 μ L/cm that microballoon, which sprays film amount,.
Drying described in dry and slitting step can be in constant temperature oven or drying chamber, and 37 DEG C dry 8-12 hours.
It is a further object to provide a kind of above-mentioned ELISA test strip apple of application, paddy, tealeaves, citrus, knots
The remaining method of metrifonate in the samples such as cabbage, Chinese cabbage, it includes step：
(1) sample pre-treatments；
(2) it is detected with test strips；
(3) testing result is analyzed.
The metrifonate time-resolved fluorescence test strips of the present invention are using the antibody antigen reaction of high degree of specificity and immune layer
Analytical technology is analysed, metrifonate monoclonal antibody-time-resolved fluorescence marker is fixed on Fusion5 films, the enemy in sample
Hundred worms are combined in flow process with metrifonate monoclonal antibody-time-resolved fluorescence marker on Fusion5 films, are formed
Drug-antibody-time-resolved fluorescence marker.Drug in sample and the metrifonate hapten-carrier in reaction film detection line
Protein conjugate competitive binding metrifonate monoclonal antibody-time-resolved fluorescence marker, final application immunochromatography detector
Calculate the metrifonate residual quantity contained in analyte sample fluid.
The test strips of the present invention have high specificity, high sensitivity, quantitative detection, at low cost, easy to operate, detection time
The advantages of short, suitable various units use, storage is simple, long shelf-life.With the remaining side of ELISA test strip metrifonate of the present invention
Method is easy, it is quick, intuitive, accurate, applied widely, at low cost, easily promote the use of.
Description of the drawings
Fig. 1 is metrifonate hapten synthesis route map.
Fig. 2 is metrifonate haptens hydrogen nuclear magnetic resonance spectrogram.
Specific implementation mode
With reference to specific embodiment, the present invention is further explained.It should be understood that these embodiments are merely to illustrate this
Invention, and be not used to limit the scope of the invention.
The preparation of 1 metrifonate test strip of embodiment
The preparation method of the test strips mainly includes the following steps that：
1) the sample absorption pad for being coated with metrifonate monoclonal antibody-time-resolved fluorescence marker is prepared；
2) preparing has the detection line for being coated with metrifonate hapten-carrier protein conjugate and is coated with anti-rabbit antibody
The reaction film of nature controlling line；
3) will test strips 1) be assembled into sample absorption pad, reaction film and the water absorption pad 2) prepared.
Substep narration in detail below：
1, the preparation of metrifonate haptens
Metrifonate 25.7g is dissolved in 100ml dichloromethane, DMAP 1g, DCC 22.66g are added, are stirred at room temperature.It is added dropwise
50ml dichloromethane solutions containing 15g p -carboxybenzaldehydes, drop are closed, and are warming up to 40 DEG C, react 6h.Solvent, column layer are removed in decompression
Analysis purifying, obtains object haptens 33g, yield 85%.By metrifonate haptens hydrogen nuclear magnetic resonance spectrogram it is found that 9.9 aldehyde
Base hydrogen, 7~8 aromatic ring hydrogen, 3.9 methylol hydrogen illustrate to synthesize successfully.
Metrifonate molecule is smaller, and is organophosphorus compound, and structural stability is poor, introduces aromatic series linking arm and strengthens
The stability of compound keeps it not degradable in immunologic process, it is easier to the high antibody of specificity be immunized out.
2, the preparation of immunogene
133mg BSA are dissolved in 5ml 0.01M PBS, stirring and dissolving.Metrifonate haptens and BSA ratios can 1: 1~
It is adjusted in the range of 60: 1, the application selects that 39mg metrifonate haptens (being 50: 1 with BSA molar ratios) is being added, and EDC is added
Hydrochloride 10mg, is stirred at room temperature 2h.The 48h that dialyses is moved into bag filter.Metrifonate immunogene is obtained under the ratio can be in antigen
Surface exposes more metrifonate haptens, to preferably cause the immune response for metrifonate.
3, the preparation of coating antigen
86mg OVA are dissolved in 5ml 0.01M PBS, stirring and dissolving.Metrifonate haptens and OVA ratios can be 1: 1~40
: it is adjusted in the range of 1, the application selects that 32mg metrifonate haptens (being 40: 1 with OVA molar ratios) is being added, and EDC salt is added
Hydrochlorate 10mg, is stirred at room temperature 2h.The 48h that dialyses is moved into bag filter.Obtained under the ratio metrifonate coating antigen with sample moderate resistance
When original competition antibody, can fully and antibody response will not be excessive because of coating antigen exposed sites to avoid false positive,
Steric hindrance is formed, to avoid false negative.
4, the preparation of metrifonate monoclonal antibody
Immunogene is injected in Balb/c new zealand white rabbit bodies, antiserum is generated.Take immune Balb/c New Zealand great Bai
Rabbit splenocyte is merged with SP2/0 myeloma cell, is screened positive hole, is obtained the hybridoma of stably excreting monoclonal antibody
Strain.Cell suspension is made with frozen stock solution in hybridoma, it is spare.Hybridoma is placed in cell culture medium, at 37 DEG C
Under the conditions of cultivated, obtained culture solution is purified with octanoic acid-saturated ammonium sulfate method, obtains monoclonal antibody, -20 DEG C
It preserves.
The cell culture medium is that calf serum and sodium bicarbonate are added into RPMI1640 culture mediums, and calf serum is made to exist
Final concentration of 20% (mass fraction) in cell culture medium, the final concentration of 0.2% (matter of sodium bicarbonate in cell culture medium
Measure score)；The pH of the cell culture medium is 7.4.It is immunogene to pathogen-free domestic using rabbit source antibody using sheep as immune animal
Sheep is immunized, and anti-rabbit antibody is obtained.
5, the preparation of microspheres solution, detection line (T lines) solution and nature controlling line (C lines) solution
(1) configuration of microspheres solution
With release buffer solution (containing 20% sucrose, 5% trehalose, 0.5%BSA, 0.2% gentamicin) by Quality Control microballoon
It is configured to time-resolved fluorescence microballoon mixed liquor, the final concentration of 0.1mg/mL-0.3mg/mL of Quality Control microballoon, detection with detection microballoon
The final concentration of 0.8mg/mL-1.2mg/mL of microballoon.
(2) preparation of detection line (T lines) solution
Metrifonate antigenic compound is diluted to 0.4mg/mL-0.6mg/mL with 0.01M PB solution.
(3) preparation of nature controlling line (C lines) solution
Streptavidin is diluted to 0.4mg/mL-0.6mg/mL with 0.01M PB solution.
6, the preparation of test strips
Reaction film is located at centre, and sample absorption pad is overlapped in reaction film left and right ends with blotting paper, is pasted onto and carries respectively
On the plastic bottom board of gum.It is 0.6 μ L/cm-1.2 μ L/cm that C, T line, which spray film liquid amount, and it is 2 μ L/cm-6 μ L/ that microspheres solution, which sprays film amount,
cm.The metrifonate for having sprayed reagent is stuck in greatly in constant temperature oven and is dried 12 hours for 37 DEG C.The metrifonate kilocalorie of drying is cut into
The paper slip of 3mm-5nm width is to get to metrifonate test strips.
The remaining detection of metrifonate in 2 sample of embodiment
1, sample treatment
(1) apple/citrus pre-treating method.
With homogenizer homogeneous samples；The apple sample after (2.00 ± 0.05) g homogeneous is weighed to centrifuge to 10mL polystyrene
Guan Zhong is separately added into 4mL 0.01M PBS, 2mL methanol, with vortex instrument whirling motion 5min；4000r/min centrifuges 5min at room temperature；
200 μ L of supernatant are taken, is added in 800 μ L 0.01M PBS, mixes well, take its 50 μ L for analyzing.
(2) paddy pre-treating method.
With homogenizer homogeneous samples；The rice sample after (2.00 ± 0.05) g homogeneous is weighed to centrifuge to 10mL polystyrene
Guan Zhong, is separately added into 10% sodium chloride of 4mL, and 2mL methanol vibrates 5min with vortex instrument；4000r/min centrifuges 5min at room temperature；
100 μ L of supernatant are taken, is added in 900 μ L0.01M PBS, mixes well, take its 50 μ L for analyzing.
(3) vegetables/tealeaves pre-treating method.
With homogenizer homogeneous samples；The vegetable sample after (1.00 ± 0.05) g homogeneous is weighed to centrifuge to 10mL polystyrene
Guan Zhong is added 2mL 0.1mol/L sulfuric acid, 5mL methanol is added, with vortex instrument whirling motion 5min；, 1000r/min centrifugations at room temperature
5min；200 μ L of supernatant are taken, is added in 800 μ L0.01M PBS, mixes well, take 50 μ L for analyzing.
2, it is detected with test strips
(1) test strips and extracting solution are restored to room temperature.
(2) 60 μ L measuring samples are accurately drawn with pipettor, (sucking bubble is careful not to when sampling) is vertically slowly added dropwise
In well (S), start timing after sample-adding, reads result within 8 minutes after sample-adding.
(3) reagent card is inserted into the carrier of immunochromatography detector, presses detection key, instrument will automatically stick into test
Row scanning.
(4) testing result is read from the display screen of immunochromatography detector.
3, testing result is analyzed
Detection limit range：1ng/ml-30ng/ml.
3 sample detection example of embodiment
1, detection limit experiment
The samples such as blank apple/paddy/tealeaves/citrus/cabbage/Chinese cabbage are taken, add metrifonate respectively to final concentration
For 0.5ng/ml, 1ng/ml, 5ng/ml, 10ng/ml, 15ng/ml, 25ng/ml, 30ng/ml, 35ng/ml, test strips is taken to carry out
Detection, each sample are repeated three times.
When with samples such as ELISA test strip apple/paddy/tealeaves/citrus/cabbage/Chinese cabbages, shown according to test strips
As a result it determines detection limit range, shows that this ELISA test strip limits range：1ng/ml-30ng/ml.
2, sample preci-sion and accuracy is tested
Using the rate of recovery as accuracy estimating index, the testing result relative standard deviation of a certain concentration samples of replication
(RSD%) it is used as precision evaluation index.Calculation formula is：The rate of recovery (%)=actual measured value/theoretical value × 100%,
Middle theoretical value is the addition concentration of sample；Relative standard deviation RSD%=SD/X × 100%, wherein SD are standard deviation, and X is
The average value of determination data.
By five concentration metrifonate of 1ng/ml, 5ng/ml, 10ng/ml, 20ng/ml, 30ng/ml respectively to apple/paddy/
The samples such as tealeaves/citrus/cabbage/Chinese cabbage be added recycling measure, each sample do 4 it is parallel, with three batches of different examinations
Agent is measured, and the average recovery rate and precision result for calculating sample see the table below.
1 sample precision of table and accuracy test
With the metrifonate of five concentration of 1ng/ml, 5ng/ml, 10ng/ml, 20ng/ml, 30ng/ml respectively to apple/rice
The samples such as paddy/tealeaves/citrus/cabbage/Chinese cabbage are added, and average recovery rate is between 83.5%~98.2%；In batch,
Relative standard deviation is respectively less than 15% between batch.
3, cross reacting rate is tested
Selection with metrifonate there are similar structures and the drug of similar functions to carry out time-resolved fluorescence test strips measurement, lead to
The standard curve for crossing various drugs respectively obtains its 50% inhibition concentration, and it is anti-to the intersection of other medicines to calculate test strips with following formula
It should rate.
2 cross reacting rate of table is tested
Claims (7)
1. a kind of time-resolved fluorescence test strips of detection metrifonate, including sample absorption pad, reaction film and water absorption pad, special
Sign is that the reaction film is located at centre, and sample absorption pad is overlapped in reaction film left and right ends respectively with water absorption pad；The sample
Absorption pad is equipped with sample application zone and microballoon area, and microballoon area is loaded with metrifonate time time-resolved fluorescence microballoon；The reaction film
It is equipped with detection line (T lines) and nature controlling line (C lines), T lines connect coating metrifonate antigen (coating antigen), and C lines are coated with anti-rabbit antibody,
The metrifonate antigen is metrifonate hapten-carrier protein conjugate, and the preparation method of the metrifonate haptens is as follows：
Metrifonate 25.7g is dissolved in 100ml dichloromethane, DMAP 1g, DCC 22.66g are added, are stirred at room temperature；Dropwise addition contains
The 50ml dichloromethane solutions of 15g p -carboxybenzaldehydes, drop are closed, and are warming up to 40 DEG C, react 6h；Decompression goes solvent, column chromatography pure
Change, obtains object haptens 33g, yield 85%.
2. the time-resolved fluorescence test strips of detection metrifonate as described in claim 1, it is characterised in that the metrifonate half
The molecular structural formula of antigen is：
3. the time-resolved fluorescence test strips of detection metrifonate as described in claim 1, it is characterised in that when the metrifonate
Between resolved fluorometric microballoon, including detection microballoon and Quality Control microballoon, detection microballoon, which is pan coating, metrifonate monoclonal antibody
Fluorescent microsphere, Quality Control microballoon are the fluorescent microsphere that pan coating has rabbit-anti labelled protein.
4. the time-resolved fluorescence test strips of detection metrifonate as described in claim 1, metrifonate monoclonal antibody is to oppose
Hundred worm hapten-carrier protein conjugates are prepared as immunogene, and the preparation method of the immunogene is as follows：
133mg BSA are dissolved in 5ml 0.01M PBS, stirring and dissolving；Metrifonate haptens and BSA ratios can be 1: 1~60: 1
In the range of adjust, the application select be added 39mg metrifonate haptens (be 50: 1 with BSA molar ratios), addition EDC hydrochloric acid
Salt 10mg, is stirred at room temperature 2h；The 48h that dialyses is moved into bag filter.
5. the time-resolved fluorescence test strips of detection metrifonate as described in claim 1, the preparation method of the coating antigen is such as
Under：
86mg OVA are dissolved in 5ml 0.01M PBS, stirring and dissolving；Metrifonate haptens and OVA ratios can be 1: 1~40: 1
Adjustment in range, the application select that 32mg metrifonate haptens (being 40: 1 with OVA molar ratios) is being added, and EDC hydrochlorides are added
2h is stirred at room temperature in 10mg；The 48h that dialyses is moved into bag filter.
6. a kind of method preparing any one of claim 1-5 test strips comprising step：
1) the sample absorption pad for being coated with metrifonate monoclonal antibody-time-resolved fluorescence marker is prepared；
2) it prepares to have and is coated with the detection line of metrifonate hapten-carrier protein conjugate and is coated with the Quality Control of anti-rabbit antibody
The reaction film of line；
3) will test strips 1) be assembled into sample absorption pad, reaction film and the water absorption pad 2) prepared.
7. the remaining method of metrifonate in the samples such as a kind of detection apple, paddy, tealeaves, citrus, cabbage, Chinese cabbage, packet
Include step：
1) Sample pretreatment；
2) it is detected with claim 1-6 any one of them test strips；
3) testing result is analyzed.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810217658.6A CN108469517A (en) | 2018-03-16 | 2018-03-16 | Time-resolved fluorescent test strip for detecting trichlorfon and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810217658.6A CN108469517A (en) | 2018-03-16 | 2018-03-16 | Time-resolved fluorescent test strip for detecting trichlorfon and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108469517A true CN108469517A (en) | 2018-08-31 |
Family
ID=63265369
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810217658.6A Pending CN108469517A (en) | 2018-03-16 | 2018-03-16 | Time-resolved fluorescent test strip for detecting trichlorfon and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108469517A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109061152A (en) * | 2018-09-21 | 2018-12-21 | 中国烟草总公司郑州烟草研究院 | A kind of time-resolved fluoroimmunoassay chromatograph test strip and its preparation method and application detecting carbendazim |
| CN113896743A (en) * | 2021-11-04 | 2022-01-07 | 深圳市易瑞生物技术股份有限公司 | Dipterex hapten, preparation method thereof, antigen, antibody and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120034711A1 (en) * | 2010-08-05 | 2012-02-09 | Oil Crops Research Institute, Chinese Academy of Agricultural Science | Digital immunochromatographic test strip for semi-quantitative detection of aflatoxin B1 and preparation method thereof |
| CN107340397A (en) * | 2017-08-29 | 2017-11-10 | 联合益康（北京）生物科技有限公司 | A kind of time-resolved fluorescence test strips for detecting cypermethrin and its application |
| CN107389927A (en) * | 2017-08-29 | 2017-11-24 | 联合益康（北京）生物科技有限公司 | A kind of time-resolved fluorescence test strips for detecting Profenofos and its application |
-
2018
- 2018-03-16 CN CN201810217658.6A patent/CN108469517A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120034711A1 (en) * | 2010-08-05 | 2012-02-09 | Oil Crops Research Institute, Chinese Academy of Agricultural Science | Digital immunochromatographic test strip for semi-quantitative detection of aflatoxin B1 and preparation method thereof |
| CN107340397A (en) * | 2017-08-29 | 2017-11-10 | 联合益康（北京）生物科技有限公司 | A kind of time-resolved fluorescence test strips for detecting cypermethrin and its application |
| CN107389927A (en) * | 2017-08-29 | 2017-11-24 | 联合益康（北京）生物科技有限公司 | A kind of time-resolved fluorescence test strips for detecting Profenofos and its application |
Non-Patent Citations (1)
| Title |
|---|
| 贲亚琍等: "农药人工抗原合成方法的设计与优选", 《农药》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109061152A (en) * | 2018-09-21 | 2018-12-21 | 中国烟草总公司郑州烟草研究院 | A kind of time-resolved fluoroimmunoassay chromatograph test strip and its preparation method and application detecting carbendazim |
| CN113896743A (en) * | 2021-11-04 | 2022-01-07 | 深圳市易瑞生物技术股份有限公司 | Dipterex hapten, preparation method thereof, antigen, antibody and application thereof |
| CN113896743B (en) * | 2021-11-04 | 2024-03-15 | 深圳市易瑞生物技术股份有限公司 | Dipterex hapten, preparation method thereof, antigen, antibody and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107340397A (en) | A kind of time-resolved fluorescence test strips for detecting cypermethrin and its application | |
| CN107907687A (en) | A kind of dicofol haptens preparation method and applications | |
| CN108535474A (en) | Time-resolved fluorescence test strip for detecting permethrin and application thereof | |
| CN105807055B (en) | A kind of test strips for detecting dichloro quinolinic acid and its preparation method and application | |
| CN109239336B (en) | Test strip for detecting isoprocarb and application thereof | |
| CN109061146B (en) | Test strip for detecting acetamiprid and preparation method and application thereof | |
| CN105785011B (en) | A kind of test strips for detecting maleic hydrazide and its preparation method and application | |
| CN104569399A (en) | Test strip for testing ochratoxin A and application of test strip | |
| CN109061147B (en) | Test strip for detecting pendimethalin and preparation method and application thereof | |
| CN109991419A (en) | Nicotinic insecticide thiacloprid method for detecting residue based on SPR technique | |
| CN109265401A (en) | A kind of preparation method and application of iprodione haptens and antigen | |
| CN101100456A (en) | Fipronil artificial hapten, synthetic method for the same, and its antigen, antibody and use | |
| CN107589265A (en) | A kind of time-resolved fluorescence test strips for detecting aflatoxin B1 and its application | |
| CN109265404A (en) | A kind of preparation method and application of carbendazim haptens and antigen | |
| CN108469517A (en) | Time-resolved fluorescent test strip for detecting trichlorfon and application thereof | |
| CN107389927A (en) | A kind of time-resolved fluorescence test strips for detecting Profenofos and its application | |
| CN112067811B (en) | Test strip for detecting alternaria alternate and application thereof | |
| CN112067814A (en) | Test strip and method for detecting difenoconazole | |
| CN108459000A (en) | Time-resolved fluorescence test strip for detecting fluopyram and application thereof | |
| CN108051583A (en) | A kind of isazofos haptens preparation method and applications | |
| CN112114147B (en) | Test strip and method for detecting pyraclostrobin | |
| CN111735951B (en) | Test strip for detecting fenpropathrin and application thereof | |
| CN109856091A (en) | Time-resolved fluorescence test strip for detecting monocrotophos and application thereof | |
| CN109265364B (en) | Preparation and application of pendimethalin hapten and antigen | |
| CN109061148B (en) | Test strip for detecting butralin and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20190523 Address after: 100144 Jinyuan Zhuang Road, Shijingshan District, Beijing 5 Applicant after: Beifang Polytechnic Univ. Address before: 100144 Jinyuan Zhuang Road, Shijingshan District, Beijing 5 Applicant before: Beifang Polytechnic Univ. Applicant before: Zhang Jing Applicant before: Wang Xiaolei |
|
| TA01 | Transfer of patent application right | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20180831 |
|
| WD01 | Invention patent application deemed withdrawn after publication |