CN111856000A - Test strip and method for detecting chlordane - Google Patents
Test strip and method for detecting chlordane Download PDFInfo
- Publication number
- CN111856000A CN111856000A CN202010499279.8A CN202010499279A CN111856000A CN 111856000 A CN111856000 A CN 111856000A CN 202010499279 A CN202010499279 A CN 202010499279A CN 111856000 A CN111856000 A CN 111856000A
- Authority
- CN
- China
- Prior art keywords
- chlordane
- pad
- conjugate
- hapten
- test strip
- 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
- BIWJNBZANLAXMG-YQELWRJZSA-N chloordaan Chemical compound ClC1=C(Cl)[C@@]2(Cl)C3CC(Cl)C(Cl)C3[C@]1(Cl)C2(Cl)Cl BIWJNBZANLAXMG-YQELWRJZSA-N 0.000 title claims abstract description 97
- 238000012360 testing method Methods 0.000 title claims abstract description 55
- 238000000034 method Methods 0.000 title claims abstract description 18
- 238000001514 detection method Methods 0.000 claims abstract description 39
- 238000006243 chemical reaction Methods 0.000 claims abstract description 32
- 239000012528 membrane Substances 0.000 claims abstract description 30
- 238000010521 absorption reaction Methods 0.000 claims abstract description 25
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 19
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 19
- 239000003550 marker Substances 0.000 claims abstract description 17
- 238000003908 quality control method Methods 0.000 claims abstract description 16
- 241000283707 Capra Species 0.000 claims abstract description 13
- 235000013305 food Nutrition 0.000 claims abstract description 10
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 9
- 229940098773 bovine serum albumin Drugs 0.000 claims description 9
- 229940092253 ovalbumin Drugs 0.000 claims description 6
- 102000014914 Carrier Proteins Human genes 0.000 claims description 5
- 108010078791 Carrier Proteins Proteins 0.000 claims description 5
- 229940124277 aminobutyric acid Drugs 0.000 claims description 5
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 claims description 5
- 230000002163 immunogen Effects 0.000 claims description 5
- 230000002745 absorbent Effects 0.000 claims description 4
- 239000002250 absorbent Substances 0.000 claims description 4
- 108010058846 Ovalbumin Proteins 0.000 claims description 3
- 230000008878 coupling Effects 0.000 claims description 3
- 238000010168 coupling process Methods 0.000 claims description 3
- 238000005859 coupling reaction Methods 0.000 claims description 3
- 238000006467 substitution reaction Methods 0.000 claims description 3
- 102000008100 Human Serum Albumin Human genes 0.000 claims description 2
- 108091006905 Human Serum Albumin Proteins 0.000 claims description 2
- 108060003552 hemocyanin Proteins 0.000 claims description 2
- 210000001685 thyroid gland Anatomy 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 1
- 238000012216 screening Methods 0.000 abstract description 5
- 238000012544 monitoring process Methods 0.000 abstract description 3
- 230000035945 sensitivity Effects 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 18
- 238000002360 preparation method Methods 0.000 description 13
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 238000011161 development Methods 0.000 description 8
- 241000699666 Mus <mouse, genus> Species 0.000 description 7
- 238000000576 coating method Methods 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- 239000011248 coating agent Substances 0.000 description 6
- 235000020185 raw untreated milk Nutrition 0.000 description 6
- 239000006143 cell culture medium Substances 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 235000020191 long-life milk Nutrition 0.000 description 5
- 235000020200 pasteurised milk Nutrition 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 210000004408 hybridoma Anatomy 0.000 description 4
- 239000008055 phosphate buffer solution Substances 0.000 description 4
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 4
- 238000005507 spraying Methods 0.000 description 4
- 239000000020 Nitrocellulose Substances 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 235000013336 milk Nutrition 0.000 description 3
- 239000008267 milk Substances 0.000 description 3
- 210000004080 milk Anatomy 0.000 description 3
- 229920001220 nitrocellulos Polymers 0.000 description 3
- 210000004989 spleen cell Anatomy 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 2
- 241001494479 Pecora Species 0.000 description 2
- 206010035226 Plasma cell myeloma Diseases 0.000 description 2
- 241000607479 Yersinia pestis Species 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 239000011230 binding agent Substances 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 244000309466 calf Species 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 230000002860 competitive effect Effects 0.000 description 2
- ZSWFCLXCOIISFI-UHFFFAOYSA-N cyclopentadiene Chemical compound C1C=CC=C1 ZSWFCLXCOIISFI-UHFFFAOYSA-N 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 230000003053 immunization Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 201000000050 myeloid neoplasm Diseases 0.000 description 2
- 238000004806 packaging method and process Methods 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 229920000728 polyester Polymers 0.000 description 2
- 229910000027 potassium carbonate Inorganic materials 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 239000001509 sodium citrate Substances 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 2
- 229940038773 trisodium citrate Drugs 0.000 description 2
- YBYIRNPNPLQARY-UHFFFAOYSA-N 1H-indene Natural products C1=CC=C2CC=CC2=C1 YBYIRNPNPLQARY-UHFFFAOYSA-N 0.000 description 1
- 241000289763 Dasygaster padockina Species 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000241125 Gryllotalpa gryllotalpa Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241000256602 Isoptera Species 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000012902 Nervous system disease Diseases 0.000 description 1
- 208000025966 Neurological disease Diseases 0.000 description 1
- 231100000674 Phytotoxicity Toxicity 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 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 1
- -1 aldrin Chemical compound 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical class N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 239000012298 atmosphere Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 150000001718 carbodiimides Chemical class 0.000 description 1
- 230000007910 cell fusion Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 229920002301 cellulose acetate Polymers 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- DFBKLUNHFCTMDC-PICURKEMSA-N dieldrin Chemical compound C([C@H]1[C@H]2[C@@]3(Cl)C(Cl)=C([C@]([C@H]22)(Cl)C3(Cl)Cl)Cl)[C@H]2[C@@H]2[C@H]1O2 DFBKLUNHFCTMDC-PICURKEMSA-N 0.000 description 1
- 229950006824 dieldrin Drugs 0.000 description 1
- NGPMUTDCEIKKFM-UHFFFAOYSA-N dieldrin Natural products CC1=C(Cl)C2(Cl)C3C4CC(C5OC45)C3C1(Cl)C2(Cl)Cl NGPMUTDCEIKKFM-UHFFFAOYSA-N 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 210000000750 endocrine system Anatomy 0.000 description 1
- RDYMFSUJUZBWLH-SVWSLYAFSA-N endosulfan Chemical compound C([C@@H]12)OS(=O)OC[C@@H]1[C@]1(Cl)C(Cl)=C(Cl)[C@@]2(Cl)C1(Cl)Cl RDYMFSUJUZBWLH-SVWSLYAFSA-N 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 238000007667 floating Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- 239000003365 glass fiber Substances 0.000 description 1
- 239000011491 glass wool Substances 0.000 description 1
- 239000003673 groundwater Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- FRCCEHPWNOQAEU-UHFFFAOYSA-N heptachlor Chemical compound ClC1=C(Cl)C2(Cl)C3C=CC(Cl)C3C1(Cl)C2(Cl)Cl FRCCEHPWNOQAEU-UHFFFAOYSA-N 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 238000003317 immunochromatography Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000000749 insecticidal effect Effects 0.000 description 1
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000014571 nuts Nutrition 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 239000003992 organochlorine insecticide Substances 0.000 description 1
- 239000003993 organochlorine pesticide Substances 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 239000000447 pesticide residue Substances 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 210000004994 reproductive system Anatomy 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000010902 straw Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000009461 vacuum packaging Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
-
- 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
- G01N33/18—Water
- G01N33/1826—Organic contamination in water
- G01N33/184—Herbicides, pesticides, fungicides, insecticides or the like
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Analytical Chemistry (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a test strip and a method for detecting chlordane. The test strip comprises a sample absorption pad, a conjugate release pad, a reaction membrane, a water absorption pad and a bottom plate, wherein the reaction membrane is provided with a detection line coated with a chlordane hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody, and the conjugate release pad is sprayed with a chlordane monoclonal antibody-colloidal gold marker. The invention also provides a method for detecting chlordane in food by applying the test strip. The test strip provided by the invention has the advantages of simple operation, high sensitivity, high detection speed, low cost, suitability for screening large-batch samples and the like, and can meet the requirements of food supervision departments in China on-site monitoring and detection.
Description
Technical Field
The invention relates to a test strip and a method for detecting chlordane, in particular to a colloidal gold test strip for detecting chlordane, which is particularly suitable for detecting residual chlordane in food.
Background
Chlordane (Chlordane) is a broad-spectrum organochlorine insecticide, also known as octachloro-hexachloro-methylene indene, and is commonly used for killing underground pests, such as mole cricket, cutworm, straw pests and the like, and has remarkable effect of controlling termites. Chlordane has contact and stomach toxicity effects on insects, has no phytotoxicity on plants under insecticidal concentration, and is toxic to mammals. Chlordane toxicity is mainly reflected in the aspects of causing disorder of a biological endocrine system, destroying a reproductive and immune system, inducing cancer and neurological diseases and the like. It is a serious environmental hazard and can permeate into the groundwater system through the surface soil, causing pollution to the water, soil and atmosphere. Therefore, it has been banned due to its high toxicity and long residue. The national standard GB 2763-2019 'maximum pesticide residue limit in food safety national standard food' specifies the residue limit of chlordane, 0.02mg/kg of vegetables, fruits, nuts and eggs and 0.002mg/kg of raw milk.
The currently reported methods for detecting chlordane mainly comprise instrument methods such as gas chromatography, gas chromatography-mass spectrometry, liquid chromatography tandem mass spectrometry and the like. The methods are operated under laboratory conditions, the sample pretreatment is complicated and time-consuming, expensive instruments and equipment are required, the detection cost is high, the time consumption is long, the operation is complex, the method has great limitation in the practical application process, and the requirement for rapidly detecting a large number of samples and field samples is difficult to meet. Therefore, the colloidal gold test strip which is simple and quick and is suitable for detecting the chlordane residue in the food is developed, the field screening and monitoring of a large number of samples can be met, and the detection work of food supervision departments and the like in China can be better met.
Disclosure of Invention
The invention aims to provide a colloidal gold test strip capable of detecting chlordane residues in food, and provides a detection method which is efficient, accurate, simple and convenient, and is suitable for field monitoring and large-scale sample screening.
The test strip for detecting chlordane provided by the invention comprises a sample absorption pad, a conjugate release pad, a reaction membrane, a water absorption pad and a bottom plate; the reaction membrane is provided with a detection line coated with a chlordane hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody; the combination release pad is sprayed with chlordane monoclonal antibody-colloidal gold marker.
The chlordane monoclonal antibody is prepared by taking a chlordane hapten-carrier protein conjugate as an immunogen.
The chlordane hapten-carrier protein conjugate is obtained by coupling chlordane hapten and carrier protein, the carrier protein is bovine serum albumin, ovalbumin, hemocyanin, thyroid protein or human serum albumin, the chlordane hapten is obtained by performing affinity substitution reaction on chlordane oxide and aminobutyric acid, and the molecular structural formula is as follows:
the sample absorption pad, the conjugate release pad, the reaction membrane and the water absorption pad are sequentially adhered to the bottom plate, and the conjugate release pads 1/3-1/2 are covered under the sample absorption pad.
The bottom plate is a PVC bottom plate or other hard non-absorbent materials; the sample absorption pad is made of polyester fiber or glass fiber; the conjugate release pad is made of glass wool or polyester material; the water absorption pad is absorbent paper; the reaction membrane is a nitrocellulose membrane or a cellulose acetate membrane.
Another object of the present invention is to provide a method for preparing the test strip, which comprises the steps of:
1) preparing a conjugate release pad sprayed with a chlordane monoclonal antibody-colloidal gold marker;
2) preparing a reaction membrane with a detection line coated with a chlordane hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody;
3) Assembling the conjugate release pad and the reaction membrane prepared in the steps 1) and 2) with a sample absorption pad, a water absorption pad and a base plate to form the test strip.
Specifically, the steps include:
1) preparing chlordane hapten through carrying out affinity substitution reaction on chlordane oxide and aminobutyric acid;
2) coupling the chlordane hapten and carrier protein to prepare a chlordane hapten-carrier protein conjugate;
3) immunizing a mouse by using the chlordane hapten-carrier protein conjugate, and fusing and screening spleen cells of the mouse and myeloma cells of the mouse to obtain a hybridoma cell strain secreting the chlordane monoclonal antibody;
4) extracting mouse IgG to immunize healthy goats to obtain goat anti-mouse anti-antibodies;
5) respectively coating the chlordane hapten-carrier protein conjugate and the goat anti-mouse anti-antibody on a detection line (T) and a quality control line (C) of a reaction membrane;
6) preparing colloidal gold by reacting trisodium citrate with chloroauric acid;
7) adding the prepared chlordane monoclonal antibody into the prepared colloidal gold to obtain a chlordane monoclonal antibody-colloidal gold marker;
8) spraying the chlordane monoclonal antibody-colloidal gold marker on a conjugate release pad, drying at 37 ℃ for 1h, taking out, and storing in a dry environment for later use;
9) soaking the sample absorption pad in 0.5% bovine serum albumin-containing phosphate buffer solution with pH of 7.2 and 0.1mol/L for 1h, and drying at 37 deg.C for more than 4 h;
10) A sample absorbing pad, a conjugate releasing pad, a reaction membrane and a water absorbing pad are sequentially adhered on the bottom plate, and the conjugate releasing pad is covered by the sample absorbing pad from the 1/3 area at the starting end. And finally cutting into small strips with the width of 3mm, adding a plastic box, carrying out vacuum packaging, and storing for 12 months at the temperature of 4-30 ℃.
The invention also aims to provide a method for detecting chlordane residue in food by using the test strip, which comprises the following steps:
(1) pretreating a sample;
(2) detecting by using a test strip;
(3) and analyzing the detection result.
The chlordane rapid detection test strip adopts a highly specific antibody-antigen reaction and immunochromatography analysis technology, a chlordane monoclonal antibody-colloidal gold marker is fixed on a conjugate release pad, chlordane in a sample is combined with the chlordane monoclonal antibody-colloidal gold marker on the conjugate release pad in the flowing process to form a drug-antibody-colloidal gold marker. The drug in the sample and the chlordane hapten-carrier protein conjugate on the reaction membrane detection line compete to combine with the chlordane monoclonal antibody-colloidal gold marker, and whether the liquid sample to be detected contains chlordane residue is judged according to the depth of a red strip of the detection line.
During detection, a sample is treated and then dropped into a test strip card hole, when the concentration of chlordane in the sample is lower than the detection limit or zero, the monoclonal antibody-colloidal gold marker is combined with a chlordane hapten-carrier protein conjugate fixed on a reaction membrane in the chromatography process, a red strip is respectively formed on a detection line (T) and a quality control line (C), and the color development of the T line is deeper than that of the C line or is consistent with that of the C line; if the concentration of chlordane in the sample is equal to or higher than the detection limit, the monoclonal antibody-colloidal gold label will bind to all of the chlordane, so that no red band will appear or the color will be lighter than that of line C at line T because the competitive reaction will not bind to the chlordane hapten-carrier protein conjugate. As shown in fig. 3.
Negative: when the quality control line (C) shows a red strip, the detection line (T) also shows a red strip, and the color of the line (T) is close to or deeper than that of the line (C), the line (C) is judged to be negative.
Positive: and when the quality control line (C) shows a red strip, the detection line (T) does not show color or the color of the line (T) is lighter than that of the line (C), the test line is judged to be positive.
And (4) invalidation: when the quality control line (C) does not show a red strip, the test strip is judged to be invalid whether the detection line (T) shows a red strip or not.
The test strip has the advantages of high sensitivity, strong specificity, low cost, simple operation, short detection time, suitability for various units, simple storage and long quality guarantee period. The method for detecting the chlordane residue by using the test strip is simple, convenient, rapid, visual, accurate, wide in application range, low in cost and easy to popularize and use.
Drawings
FIG. 1 is a diagram of synthesis of chlordane hapten.
FIG. 2 is a schematic cross-sectional view of a test strip.
FIG. 3 is a diagram showing the test result of the test strip.
Detailed Description
The invention is further illustrated below with reference to specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.
Example 1 preparation of test strip for detecting chlordane
The preparation method of the test strip mainly comprises the following steps:
1) preparing a conjugate release pad sprayed with a chlordane monoclonal antibody-colloidal gold marker;
2) preparing a reaction membrane with a detection line coated with a chlordane hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody;
3) assembling the conjugate release pad and the reaction membrane prepared in the steps 1) and 2) with a sample absorption pad, a water absorption pad and a base plate to form the test strip.
The following steps are detailed:
1. synthesis of chlordane hapten (the synthetic route is shown in figure 1)
Dissolving 0.423g of chlordane oxide in 100mL of methanol; dissolving 0.206g of aminobutyric acid in 5mL of water, adding the dissolved aminobutyric acid into a methanol solution of chlordane oxide, and fully stirring; dissolving 0.43g of potassium carbonate in 5mL of water, adding the dissolved potassium carbonate into the reaction solution, stirring the mixture at 55 ℃ for 5 hours, stopping the reaction, adding 6mol/L hydrochloric acid to adjust the pH value to 6, performing rotary evaporation and concentration to remove methanol, adding 50mL of water and 100mL of chloroform, extracting the mixture for three times, combining organic phases, evaporating the mixture to dryness to obtain a yellow oily substance, adding 12mL of absolute ethyl alcohol, and recrystallizing to obtain the chlordane hapten.
2. Preparation of immunogens
Taking 19.6mg of chlordane hapten, adding 1mL of N, N-Dimethylformamide (DMF) for dissolving, adding 13.3mg of N-hydroxysuccinimide (NHS) and 15.7mg of carbodiimide (EDC), and reacting for 3h at room temperature to obtain a hapten solution A; taking 50mg of Bovine Serum Albumin (BSA), and adding 0.05mol/L CB buffer solution for dissolving to obtain solution B; dripping the A solution into the B solution, reacting at 4 deg.C for 12h, dialyzing and purifying with 0.02mol/L PBS for 3 days, changing solution 3 times per day to obtain chlordane hapten-BSA conjugate as immunogen, packaging, and storing at-20 deg.C.
3. Preparation of coating antigen
Taking 12mg of chlordane hapten, adding 1mL of dimethyl sulfoxide (DMSO) for dissolving, adding 8.4mg of NHS and 10.2mg of EDC, and reacting at room temperature for 3h to obtain a hapten solution A; dissolving Ovalbumin (OVA)50mg in CB buffer solution of 0.05mol/L to obtain solution B; dripping the A solution into the B solution, reacting at 4 deg.C for 12h, dialyzing and purifying with 0.02mol/L PBS for 3 days, changing solution 3 times per day to obtain chlordane hapten-OVA conjugate, packaging, and storing at-20 deg.C.
4. Preparation of chlordane monoclonal antibody
(1) Animal immunization
Injecting the immunogen obtained in the step 2 into Balb/c mice at an immune dose of 150 mug/mouse to generate antiserum.
(2) Cell fusion and cloning
Taking immune Balb/c mouse spleen cells, fusing the immune Balb/c mouse spleen cells with SP2/0 myeloma cells according to the proportion of 8:1 (quantitative ratio), measuring cell supernatant by adopting indirect competitive ELISA, and screening positive holes. Cloning the positive hole by using a limiting dilution method until obtaining the hybridoma cell strain which stably secretes the monoclonal antibody.
(3) Cell cryopreservation and recovery
Making hybridoma cell into 1 × 10 with frozen stock solution6Cell suspension per mL, preserved for long period in liquid nitrogen. Taking out the freezing tube during recoveryImmediately placing the mixture into a water bath at 37 ℃ for quick melting, centrifuging to remove a frozen stock solution, and transferring the frozen stock solution into a culture bottle for culture.
(4) Preparation and purification of monoclonal antibodies
An incremental culture method: placing the hybridoma cell in cell culture medium, culturing at 37 deg.C, purifying the obtained culture solution by octanoic acid-saturated ammonium sulfate method to obtain monoclonal antibody, and storing at-20 deg.C.
The cell culture medium is prepared by adding calf serum and sodium bicarbonate into RPMI1640 culture medium to make the final concentration of calf serum in the cell culture medium 20% (mass fraction) and the final concentration of sodium bicarbonate in the cell culture medium 0.2% (mass fraction); the pH of the cell culture medium was 7.4.
5. Preparation of goat anti-mouse anti-antibody
The sheep is taken as an immune animal, and the pathogen-free sheep is immunized by taking the murine antibody as an immunogen to obtain the goat anti-mouse antibody.
6. Preparation of chlordane monoclonal antibody-colloidal gold marker
(1) Preparation of colloidal gold
Diluting 1% chloroauric acid to 0.01% (mass fraction) with double distilled deionized water, placing 100mL into a conical flask, heating to boil with a constant temperature electromagnetic stirrer, adding 1.5mL of 1% trisodium citrate under continuous stirring at high temperature, stirring and heating at constant speed until the solution is bright red, cooling to room temperature, recovering the volume with deionized water, and storing at 4 deg.C. The prepared colloidal gold has pure appearance, is transparent and bright, has no sediment or floating objects, and is wine red when observed in sunlight.
(2) Preparation of chlordane monoclonal antibody-colloidal gold marker
Under magnetic stirring, adjusting the pH value of the colloidal gold to 7.2 by using 0.2mol/L potassium carbonate solution, adding the chlordane monoclonal antibody into the colloidal gold solution according to the standard that 5-50 mu g of antibody is added into each milliliter of the colloidal gold solution, and continuously stirring and uniformly mixing for 30 min; after standing for 10min, 10% BSA was added to make the final concentration of the solution in colloidal gold 1%, and the solution was allowed to stand for 10 min. Centrifuging at 12000r/min at 4 deg.C for 40min, discarding supernatant, washing the precipitate with redissolving buffer twice, resuspending the precipitate with redissolving buffer whose volume is 1/10 of the original volume of colloidal gold, and standing at 4 deg.C for use.
Redissolving buffer solution: 0.02mol/L phosphate buffer solution containing 0.1-0.5 percent of BSA, 2-4 percent of sucrose and pH7.2.
7. Preparation of conjugate Release pad
The conjugate release pad was soaked in 0.5% BSA, pH7.2, 0.5mol/L phosphate buffer, soaked for 1h, and baked at 37 deg.C for 3 h. And uniformly spraying the prepared chlordane monoclonal antibody-colloidal gold marker on a conjugate release pad by using an Isoflow film spraying instrument, spraying 0.01mL of the chlordane monoclonal antibody-colloidal gold marker on every 1cm of the conjugate release pad, placing the mixture in an environment at 37 ℃ (the humidity is less than 20%) for 60min, taking out the mixture, and placing the mixture in a dry environment (the humidity is less than 20%) for storage for later use.
8. Preparation of sample absorbent pad
The sample absorption pad is soaked in 0.1mol/L phosphate buffer solution containing 0.5 percent of bovine serum albumin and having the pH value of 7.2 for 1h, and is dried for more than 4h at the temperature of 37 ℃ for later use.
9. Preparation of the reaction film
Coating the chlordane hapten-ovalbumin conjugate on a reaction membrane to form a detection line, and coating the goat anti-mouse anti-antibody on the reaction membrane to form a quality control line.
Coating process: diluting the chlordane hapten-ovalbumin conjugate to 1mg/mL by using 0.01mol/L, pH 7.4.4 phosphate buffer solution, and coating the chlordane hapten-ovalbumin conjugate on a detection line (T line) on a nitrocellulose membrane by using an Isoflow point membrane instrument, wherein the coating amount is 1.0 mu L/cm; the goat anti-mouse anti-antibody was diluted to 200. mu.g/mL with 0.01mol/L, pH 7.4.4 phosphate buffer and coated on a quality control line (line C) on a nitrocellulose membrane in an amount of 1.0. mu.L/cm using an Isoflow dot membrane apparatus. And (3) drying the coated reaction membrane for 2 hours at 37 ℃ for later use.
10. Assembly of test strips
According to the section structure of the test strip shown in the attached figure 2, a sample absorption pad (1), a conjugate release pad (2), a reaction membrane (3) and a water absorption pad (4) are sequentially adhered to a PVC bottom plate (7); the binder release pad is covered by the sample absorption pad from the 1/3 area at the starting end, the tail end of the binder release pad is connected with the starting end of the reaction film, the tail end of the reaction film is connected with the starting end of the water absorption pad, the starting end of the sample absorption pad is aligned with the starting end of the PVC base plate, and the tail end of the water absorption pad is aligned with the tail end of the PVC base plate; the reaction membrane is provided with a detection line (5) and a quality control line (6), and the detection line (T line) and the quality control line (C line) are strip-shaped strips which are vertical to the long phase of the test strip; the detection line is located on the side near the end of the conjugate release pad; the quality control line is positioned on the side away from the end of the conjugate release pad; cutting the test strip into small strips with the width of 3mm by a machine, putting the small strips into a special plastic card, and storing the small strips in an environment with the temperature of 4-30 ℃ for 12 months.
EXAMPLE 2 detection of chlordane in milk
1. Sample pretreatment
And (4) restoring the milk sample to room temperature for detection.
2. Detection with test strips
Sucking 80 mu L of sample liquid to be detected by a micropipette and vertically dripping the sample liquid into the sample adding hole; the liquid flow was started, the reaction was carried out for 10min, and the results were judged.
3. Analyzing the results of the detection
Negative (-): the color development of the T line is darker than that of the C line or is consistent with that of the C line, which indicates that the concentration of chlordane in the sample is lower than the detection limit, as shown in FIGS. 3a and 3 b.
Positive (+): the color development of the T line is lighter than that of the C line or the T line is not developed, which indicates that the chlordane concentration in the sample is equal to or higher than the detection limit, as shown in FIGS. 3C and 3 d.
And (4) invalidation: the absence of line C indicates an incorrect procedure or the test strip has deteriorated and failed, as shown in FIGS. 3e and 3 f.
Example 3 sample testing example
1. Limit of detection test
Taking blank raw milk, pasteurized milk and UHT sterilized milk samples, respectively adding chlordane to the samples with final concentrations of 1 mug/L, 2 mug/L and 4 mug/L, taking test strips for detection, and repeatedly measuring each sample for three times.
When the test paper is used for detecting raw milk, pasteurized milk and UHT sterilized milk samples, when no chlordane exists and the addition concentration of the chlordane is 1 mu g/L, the test paper shows that the color development of a T line is darker than that of a C line or is consistent with that of the C line, and the test paper is negative; when the concentration of the chlordane added is 2 mug/L and 4 mug/L, the test strip shows that the color development of a T line is lighter than that of a C line or the color development of the T line is not shown, and the test strip is positive, which shows that the detection limit of the test strip on chlordane in raw milk, pasteurized milk and UHT sterilized milk is 2 mug/L, and meets the requirement of residual limit in GB 2763.
2. Test for false positive and false negative rates
Taking blank raw milk, pasteurized milk and UHT sterilized milk samples and 20 parts of positive raw milk, pasteurized milk and UHT sterilized milk samples added with chlordane to the final concentration of 2 mug/L, respectively detecting by using test strips produced by 3 batches, and calculating the negative and positive rates.
The results show that: when 3 batches of test strips are used for detecting positive samples, the results are all positive, the positive coincidence rate is 100 percent, and the false negative rate is 0; when negative samples are detected, the results are all negative, and the negative coincidence rate is 100 percent and the false positive rate is 0. The test strip for detecting chlordane can be used for quickly detecting the chlordane residue in milk.
3. Specificity test
When the test strip is used for detecting other cyclopentadiene organochlorine pesticides such as 10mg/L endosulfan, dieldrin, aldrin, heptachlor and the like, the test strip shows that the T line color development is darker than or consistent with the C line color development and is negative, which indicates that the test strip has no cross reaction to the drugs.
Claims (5)
1. A test strip for detecting chlordane comprises a sample absorption pad, a conjugate release pad, a reaction membrane, a water absorption pad and a bottom plate, wherein the reaction membrane is provided with a detection line coated with a chlordane hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody, and the conjugate release pad is sprayed with a chlordane monoclonal antibody-colloidal gold marker; the chlordane monoclonal antibody is prepared by taking a chlordane hapten-carrier protein conjugate as an immunogen; the chlordane hapten-carrier protein conjugate is obtained by coupling chlordane hapten and carrier protein, wherein the carrier protein is bovine serum albumin, ovalbumin, hemocyanin, thyroid protein or human serum albumin, and is characterized in that the chlordane hapten is obtained by performing affinity substitution reaction on chlordane oxide and aminobutyric acid, and the molecular structural formula of the chlordane hapten-carrier protein conjugate is as follows:
2. The strip of claim 1, wherein the sample absorbing pad, the conjugate releasing pad, the reaction membrane, and the absorbent pad are sequentially attached to the base plate.
3. The strip of any one of claims 1-2, wherein the conjugate release pad 1/3-1/2 is covered under a sample absorbing pad.
4. A method of making the test strip of any one of claims 1-3, comprising the steps of:
1) preparing a conjugate release pad sprayed with a chlordane monoclonal antibody-colloidal gold marker;
2) preparing a reaction membrane with a detection line coated with a chlordane hapten-carrier protein conjugate and a quality control line coated with a goat anti-mouse anti-antibody;
3) assembling the conjugate release pad and the reaction membrane prepared in the steps 1) and 2) with a sample absorption pad, a water absorption pad and a base plate to form the test strip.
5. A method for detecting chlordane residue in food comprises the following steps:
1) pretreating a sample;
2) performing a test using the test strip of any one of claims 1-3;
3) and analyzing the detection result.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010499279.8A CN111856000B (en) | 2020-06-04 | 2020-06-04 | Test strip and method for detecting chlordane |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010499279.8A CN111856000B (en) | 2020-06-04 | 2020-06-04 | Test strip and method for detecting chlordane |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111856000A true CN111856000A (en) | 2020-10-30 |
| CN111856000B CN111856000B (en) | 2023-07-07 |
Family
ID=72985485
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010499279.8A Active CN111856000B (en) | 2020-06-04 | 2020-06-04 | Test strip and method for detecting chlordane |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111856000B (en) |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992012427A1 (en) * | 1991-01-08 | 1992-07-23 | United States Department Of Energy | Monoclonal antibodies to cyclodiene insecticides and method for detecting the same |
| US5334528A (en) * | 1989-10-30 | 1994-08-02 | The Regents Of The University Of California | Monoclonal antibodies to cyclodiene insecticides and method for detecting the same |
| US20030068786A1 (en) * | 2001-10-10 | 2003-04-10 | Rani Bangalore Eahwar Amita | Process for the production of egg yolk antibodies for organochlorine pesticides |
| US20110159519A1 (en) * | 2007-01-24 | 2011-06-30 | Carnegie Mellon University | Optical biosensors |
| CN102701972A (en) * | 2012-04-27 | 2012-10-03 | 湖南大学 | Octachlorostyrene hapten, artificial antigen and specific antibody |
| CN105785011A (en) * | 2016-03-15 | 2016-07-20 | 中国烟草总公司郑州烟草研究院 | Test strip for detecting maleic hydrazide, as well as preparation method and application thereof |
| CN106324263A (en) * | 2015-07-01 | 2017-01-11 | 北京安达兴业科技有限公司 | Test strip for detecting estriol and applications thereof |
| CN110850090A (en) * | 2019-10-25 | 2020-02-28 | 成都市房屋安全事务中心 | Test strip for detecting bifenthrin and application thereof |
-
2020
- 2020-06-04 CN CN202010499279.8A patent/CN111856000B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5334528A (en) * | 1989-10-30 | 1994-08-02 | The Regents Of The University Of California | Monoclonal antibodies to cyclodiene insecticides and method for detecting the same |
| WO1992012427A1 (en) * | 1991-01-08 | 1992-07-23 | United States Department Of Energy | Monoclonal antibodies to cyclodiene insecticides and method for detecting the same |
| US20030068786A1 (en) * | 2001-10-10 | 2003-04-10 | Rani Bangalore Eahwar Amita | Process for the production of egg yolk antibodies for organochlorine pesticides |
| US20110159519A1 (en) * | 2007-01-24 | 2011-06-30 | Carnegie Mellon University | Optical biosensors |
| CN102701972A (en) * | 2012-04-27 | 2012-10-03 | 湖南大学 | Octachlorostyrene hapten, artificial antigen and specific antibody |
| CN106324263A (en) * | 2015-07-01 | 2017-01-11 | 北京安达兴业科技有限公司 | Test strip for detecting estriol and applications thereof |
| CN105785011A (en) * | 2016-03-15 | 2016-07-20 | 中国烟草总公司郑州烟草研究院 | Test strip for detecting maleic hydrazide, as well as preparation method and application thereof |
| CN110850090A (en) * | 2019-10-25 | 2020-02-28 | 成都市房屋安全事务中心 | Test strip for detecting bifenthrin and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| B. E. AMITA RANI 等: "Immunoanalysis of Endosulphan Residues in Spices", 《FOOD AND AGRICULTURAL IMMUNOLOGY》, vol. 15, no. 2 * |
| JUAN J. MANCLUÄS 等: "Development of a Monoclonal Immunoassay Selective for Chlorinated Cyclodiene Insecticides", 《JOURNAL AGRICULTURAL AND FOOD CHEMISTRY》, vol. 52, no. 10 * |
| KIRK W. JOHNSON 等: "An immunotoxicological evaluation of γ-chlordane", 《FUNDAMENTAL AND APPLIED TOXICOLOGY》, vol. 6, no. 2, XP024877266, DOI: 10.1016/0272-0590(86)90246-0 * |
| 王振国 等: "食品农药残留免疫学技术检测", 《吉林农业大学学报》, vol. 19, no. 3 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111856000B (en) | 2023-07-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111239399B (en) | Test strip and method for detecting profenofos | |
| CN110850090A (en) | Test strip for detecting bifenthrin and application thereof | |
| CN108776219B (en) | Immunochromatographic test strip for rapidly detecting alternaria tenuipili acid | |
| CN110441518B (en) | Test strip and method for detecting gibberellin | |
| CN109239336B (en) | Test strip for detecting isoprocarb and application thereof | |
| CN107271665B (en) | Test strip for detecting salbutamol and application thereof | |
| CN106918705B (en) | Test paper for detecting fenpropathrin and application thereof | |
| CN112067811B (en) | Test strip for detecting alternaria alternate and application thereof | |
| CN109682961B (en) | Test strip for detecting cyphenothrin and application thereof | |
| CN112067814A (en) | Test strip and method for detecting difenoconazole | |
| CN108931640B (en) | Test strip for detecting organophosphorus pesticide and application thereof | |
| CN110551220A (en) | Preparation and application of DDT monoclonal antibody | |
| CN113156125B (en) | Test strip and method for detecting milbemycetin | |
| CN111735951B (en) | Test strip for detecting fenpropathrin and application thereof | |
| CN111965360B (en) | Test strip and method for detecting procymidone | |
| CN111751535B (en) | Test strip for detecting endosulfan and application thereof | |
| CN113267622B (en) | Test strip and method for detecting chlorohydroxypyridine | |
| CN112114147B (en) | Test strip and method for detecting pyraclostrobin | |
| CN110441517B (en) | Test strip and method for detecting 6-benzyladenine | |
| CN111856000B (en) | Test strip and method for detecting chlordane | |
| CN111289752A (en) | Test strip and method for detecting dicofol | |
| CN111896738A (en) | Test strip for detecting serpentrin and application thereof | |
| CN111965359B (en) | Test strip and method for detecting chlorfenapyr | |
| CN111735952B (en) | Test strip and method for detecting hexachlorobenzene | |
| CN113156127B (en) | Test strip and method for detecting chlorpyrifos |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: No.8, Gaoxin 4th Street, Huilongguan international information industry base, Changping District, Beijing Applicant after: Beijing Qinbang Technology Co.,Ltd. Applicant after: TANGSHAN FOOD AND DRUG COMPREHENSIVE INSPECTION AND TESTING CENTER Address before: No.8, Gaoxin 4th Street, Huilongguan international information industry base, Changping District, Beijing Applicant before: BEIJING KWINBON BIOTECHNOLOGY Co.,Ltd. Applicant before: TANGSHAN FOOD AND DRUG COMPREHENSIVE INSPECTION AND TESTING CENTER |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |