CN106610432A - Enzyme-linked immunosorbent assay kit for detecting methomyl and detection method thereof - Google Patents

Enzyme-linked immunosorbent assay kit for detecting methomyl and detection method thereof Download PDF

Info

Publication number
CN106610432A
CN106610432A CN201510694322.5A CN201510694322A CN106610432A CN 106610432 A CN106610432 A CN 106610432A CN 201510694322 A CN201510694322 A CN 201510694322A CN 106610432 A CN106610432 A CN 106610432A
Authority
CN
China
Prior art keywords
methomyl
solution
enzyme
preparation
detection
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.)
Withdrawn
Application number
CN201510694322.5A
Other languages
Chinese (zh)
Inventor
洪霞
杜霞
张淑雅
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jiangsu Wise Science and Technology Development Co Ltd
Original Assignee
Jiangsu Wise Science and Technology Development Co Ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Jiangsu Wise Science and Technology Development Co Ltd filed Critical Jiangsu Wise Science and Technology Development Co Ltd
Priority to CN201510694322.5A priority Critical patent/CN106610432A/en
Publication of CN106610432A publication Critical patent/CN106610432A/en
Withdrawn legal-status Critical Current

Links

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/577Immunoassay; 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)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Cell Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Food Science & Technology (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The invention discloses an enzyme-linked immunosorbent assay kit for detecting methomyl and a detection method thereof. The kit realizes sensitive, accurate and rapid detection, can be simply operated, has strong specificity and is suitable for detection of a large batch of samples. The kit comprises an enzyme-labeled plate coated with a methomyl antigen, a methomyl standard product, a methomyl antibody working solution, a methomyl enzyme-labeled secondary antibody working solution, a substrate liquid A, a substrate liquid B, a stop solution, a concentration diluent and a concentration washing solution. The principle of the enzyme-linked immunosorbent assay kit for detection of methomyl is solid phase indirect competition enzyme-linked immunosorbent assay. The detection method comprises adding an extracted sample, the enzyme-labeled secondary antibody working solution and the antibody working solution into corresponding micropores of the enzyme-labeled plate, carrying out incubation for some time, washing the enzyme-labeled plate, adding the substrate liquids A and B into the enzyme-labeled plate, carrying out development under action of the enzyme so that the development agent is blue, and adding the stop solution into the sample so that the blue color becomes yellow color, wherein the depth of the color is inversely proportional with the methomyl content of the standard substance or sample. The method can be directly used for detecting residual quantity of methomyl in a detected crop.

Description

A kind of enzyme linked immunological kit and its detection method of detection Methomyl
Technical field
The present invention relates to detection of veterinary drugs in food technical field, particularly detects the enzyme linked immunological kit of Methomyl in rice.
Background technology
Methomyl is a kind of broad spectrum activity carbamate insecticides, with characteristics such as high volatility, toxicity on inhalation height, is mainly used in preventing and treating the class pests such as striped rice borer, plant hopper, prodenia litura.Active compound is white crystalline powder, and commodity state is wettable powder, soluble powder or missible oil.Methomyl be a kind of absorbability have tag, the broad spectrum pesticide of the carbamates of stomach poison function.Recommend at first as desinsection, nematicide.Suitable for cotton, tobacco, fruit tree, vegetables it is anti-eliminate aphis, moth, cutworm etc..Therefore it is regular that methomyl residue is carried out to detect the necessary means of the monitoring for becoming many countries.
Enzyme linked immunosorbent assay is a kind of accurate, reliable, quick, special detection method, is suitable for the quick screening of gross sample, and food safety detection industry is widely used in recent years.It is contemplated that setting up a kind of enzyme linked immunological kit and its detection method of detection Methomyl.
The content of the invention
In order to overcome chromatographic deficiency, the present invention to provide a kind of enzyme linked immunological kit and its detection method of detection Methomyl.The method is sensitive, accurate, quick, easy to operate, high specificity, it is adaptable to the quick detection of gross sample.
The enzyme linked immunological kit of present invention detection Methomyl, including ELISA Plate, Methomyl standard items, Methomyl antibody working solution, Methomyl ELIAS secondary antibody working solution, substrate solution A, substrate solution B, terminate liquid, concentration and dilution liquid and concentrated cleaning solution.
The preparation of the enzyme linked immunological kit of present invention detection Methomyl, comprises the following steps:Preparation, the preparation of Methomyl standard items, the preparation of Methomyl antibody working solution, the preparation of Methomyl ELIAS secondary antibody working solution, the preparation of substrate solution A, the preparation of substrate solution B, the preparation of terminate liquid, the preparation of concentration and dilution liquid and the preparation of concentrated cleaning solution of ELISA Plate.
It is further characterized by:Described ELISA Plate is prepared via Methomyl antigen coat, comprise the concrete steps that to be coupled Methomyl haptens and the pure albumen of carrier proteins Bovine (BSA) and obtain envelope antigen, with carbonate (CBS) buffer solution of 0.05 mol/L pH 9.6 as coating buffer, Methomyl envelope antigen is diluted to into 1:40000 ratios, 100 μ L/ holes, 37 DEG C of lucifuges are incubated 2 h, take out ELISA Plate and get rid of liquid in plate, add the μ L/ holes of diluted concentrated cleaning solution 300, board-washing 2 times, 30 s/ time, it is subsequently adding 0.5% bovine serum albumin(BSA) (BSA) confining liquid, 150 μ L/ holes, 37 DEG C of 1.5 h of placement, get rid of confining liquid and directly pat dry, ELISA Plate after patting dry is placed (25 DEG C) between constant temperature and is dried, and preserves under the conditions of ELISA Plate is vacuum-sealed in into 4 DEG C after sampling observation is qualified.
Methomyl standard concentration is respectively 0 mg/kg, 0.1mg/kg, 0.3 mg/kg, 0.9 mg/kg, 2.7 mg/kg, 8.1 mg/kg.
The Methomyl antibody working solution is to obtain monoclonal antibody using Methomyl artificial antigen immune mouse, and with antibody diluent 1 is diluted to:It is prepared by 60000 ratios.
The Methomyl ELIAS secondary antibody working solution adds diluted into 1 by ELIAS secondary antibody:2000 ratios, the substrate solution A is the citrate-phosphate disodium hydrogen cushioning liquid of the carbamide peroxide containing 0.5 mmol/L, and the substrate solution B is the ethanol solution of tetramethyl biphenyl diamines, and the terminate liquid is 2 The sulfuric acid of mol/L, the concentration and dilution liquid is 10 times of concentration and dilution liquid, is the PBS of 0.1 mol/L, between pH value range 7.0-7.5, the concentrated cleaning solution is 10 times of concentrated cleaning solutions, is containing 0.5% Tween-20, the PBST of 0.1 mol/L, between pH value range 7.0-7.5.
The enzyme linked immunological kit and its detection method of detection Methomyl, based on the indirect competitive enzyme-linked immunosorbent reaction principle of antigen-antibody, the method is comprised the following steps:
(1) testing sample is pre-processed, sample treatment that will be to be tested is fluid sample, or uses Solvent Extract methods testing sample, and is redissolved in sample diluting liquid;
(2) required reagent is taken out from cold storage environment, is placed in room temperature(20~25 DEG C)30 more than min are balanced, notices that every kind of liquid reagent must shake up using before;
(3) ELISA Plate for being coated with Methomyl antigen is taken, plus the μ L/ holes of standard items/sample 50 are in corresponding micropore, add Methomyl ELIAS secondary antibody working solution, 50 μ L/ holes, it is subsequently adding Methomyl antibody working solution, 50 μ L/ holes, gently vibration is mixed, and with cover plate membrane cover plate 25 DEG C of light protected environments of rearmounted room temperature 30 min are reacted;
(4) cover plate film is carefully opened, liquid in hole is dried, with the μ L/ holes of wash operating solution 260, fully washed 4 ~ 5 times, per the s of minor tick 10, patted dry (bubble not being eliminated after patting dry can be poked with original pipette tips) with blotting paper;
(5) the μ L/ holes of substrate solution A 50, the μ L/ holes of substrate solution B 50, gently vibration is added to mix, with 15~20 min of reaction in the rearmounted 25 DEG C of light protected environments of cover plate membrane cover plate;
(6) the μ L/ holes of terminate liquid 50 are added, gently vibration is mixed, setting ELIASA is at 450 nm or the nm of dual wavelength 450/630 is detected, determines per hole absorbance(Please run through data in 5 min);
(7) with standard concentration (ppb) logarithm as abscissa, standard items percentage absorbance be ordinate, draw calibration curve, reference standard curve calculate sample in Methomyl content.
The enzyme linked immunological kit of present invention detection Methomyl and its measuring principle of detection method:Methomyl in sample and antigen-specific sexual competition antibody fixed on ELISA Plate, add ELIAS secondary antibody, and antibody response, by enzymatic chromogenic reagent, according to the depth of colour developing come the content of Methomyl in judgement sample.If the Methomyl Content in sample is few, colour developing is deep;Conversely, it is shallow then to develop the color.The kit test method of the present invention is easy to operate, detects sensitive, accurate, quick, it is adaptable to the quick detection of batch samples.
Specific embodiment
The preparation of Methomyl protein conjugate:
Methomyl hapten derivant with carboxyl is obtained using succinyl oxide method, 0.05 mmol and carrier protein BSA is taken afterwards by 10:1 combination ratio is blended in 0.05 In the carbonate buffer solution (CBS) of mol/L pH 9.6, it is subsequently adding 0.15 mmol carbodiimides, the h of room temperature reaction 24 is put in stirring, dialyse two days most in the PBS of 0.2 mol/L pH 7.6, unreacted haptens is removed, the protein conjugate solution for obtaining is saved backup in -20 DEG C.
The preparation of Methomyl antibody:
From the purebred BALA/C mouse of healthy adult, take to mix with equivalent complete Freund's adjuvant with the μ g of immunizing antigen 50 of protein molecule preparation and initial immunity is carried out using lumbar injection, equivalent incomplete Freund's adjuvant was added to carry out using lumbar injection with same dose immunizing antigen every 3 weeks afterwards secondary, three immunity, every time tail vein blood determines antiserum titre to certain titre after immunity 6 days, adjuvant is not added with same dose carries out final immunization, spleen is taken after 3 days preparing Spleen cell suspensions and myeloma cell carries out cell fusion, hybridoma cell line required for filtering out carries out cloning, the hybridoma in exponential phase is selected to carry out frozen, for ascites preparation, first lumbar injection 0.5 Ml atoleines are in BALB/C mouse sensitization, pneumoretroperitoneum injection 1 × 10 in 2 weeks6Individual hybridoma, inoculating cell can produce ascites after 7-10 days, ascites is extracted with syringe when ascites is as more as possible, collect repeatedly for several times, 4000 Rpm is centrifuged 15 min, collects supernatant, and monoclonal antibody is purified using caprylic acid-ammonium purifying ascites, and freeze-drying obtains freeze-dried powder and saves backup after -20 DEG C.
Preparation is coated with the ELISA Plate of Methomyl envelope antigen:
Methomyl haptens and carrier proteins Bovine pure albumen (BSA) are coupled and are obtained by envelope antigen, with carbonate (CBS) buffer solution of 0.05 mol/L pH 9.6 as coating buffer, by Methomyl antigen diluent into 1:40000 ratios, 100 L/ holes, 37 DEG C of 2 h of placement take out ELISA Plate and get rid of liquid in plate, and with the L/ holes of concentrated cleaning solution 300 after dilution, board-washing 2 times 30 s/ time, is subsequently adding 0.5% bovine serum albumin(BSA) (BSA) and closes, and 150 L/ holes, 37 DEG C of 1.5 h of placement, discard confining liquid, and ELISA Plate after patting dry is placed (25 DEG C) between constant temperature and dried, inspect by random samples it is qualified after by ELISA Plate vacuum sealing it is rearmounted 4 DEG C at preserve.
Methomyl standard items compound concentration is respectively 0 mg/kg, 0.1mg/kg, 0.3 mg/kg, 0.9 mg/kg, 2.7 mg/kg, 8.1 mg/kg.
The preparation of Methomyl antibody working solution:Monoclonal antibody is obtained using Methomyl artificial antigen immune mouse, with antibody diluent 1 is diluted to:It is prepared by 60000 ratios.
Methomyl ELIAS secondary antibody working solution adds diluted into 1 by ELIAS secondary antibody:2000 ratios, substrate solution A is the citrate-phosphate disodium hydrogen cushioning liquid of the carbamide peroxide containing 0.5 mmol/L, and substrate solution B is the ethanol solution of tetramethyl biphenyl diamines, and terminate liquid is 2 The sulfuric acid of mol/L, concentration and dilution liquid is 10 times of concentration and dilution liquid, is the PBS of 0.1 mol/L, and between pH value range 7.0-7.5, concentrated cleaning solution is 10 times of concentrated cleaning solutions, is containing 0.5% Tween-20, the PBST of 0.01mol/L, between pH value range 7.0-7.5.
Based on the reagent of above-mentioned preparation, the present invention includes following material for the enzyme linked immunological kit for detecting Methomyl:
(1) 96 hole elisa Plates × 1 piece;
(2) titer × 6 bottle:(1mL/ bottles) 0 mg/kg, 0.1mg/kg, 0.3 mg/kg, 0.9 mg/kg, 2.7 mg/kg, 8.1 mg/kg;
(3) mL of antibody working solution 7;
(4) mL of ELIAS secondary antibody working solution 7;
(5) mL of substrate solution A 7;
(6) mL of substrate solution B 7;
(7) mL of terminate liquid 7;
The mL of (8) 10 × concentration and dilution liquid 40;
The mL of (9) 10 × concentrated cleaning solution 40;
When the kit of the present invention is used to detect methomyl residue amount in crop sample, implemented by following steps:Sample pretreatment, detected with kit of the present invention, analysis result.
(1) with methomyl residue amount in kit of the present invention detection testing sample
The ELISA Plate for being coated with Methomyl antigen is taken, plus the L/ holes of standard items/sample 50 are in corresponding micropore;ELIAS secondary antibody working solution, 50 L/ holes is added to be subsequently adding Methomyl antibody working solution, 50 L/ holes, gently vibration is mixed, and with cover plate membrane cover plate 25 DEG C of light protected environments of rearmounted room temperature 30 min are reacted;Cover plate film is carefully opened, liquid in hole is dried, with wash operating solution 300 L/ holes, fully washing 4 times, soak 15-30 s, are patted dry with blotting paper;Add the L/ holes of substrate solution A 50, the L/ holes of substrate solution B 50, gently vibration to mix, with the rearmounted 25 DEG C of light protected environments of cover plate membrane cover plate 15 min are reacted;The L/ holes of terminate liquid 50 are added, gently vibration is mixed, setting ELIASA is in 450 At nm or the nm of dual wavelength 450/630 detections, determine per hole absorbance(Please run through data in 5 min);The absorbance size of contrast testing sample and standard items, methomyl residue amount in quantitative analysis testing sample.
(2) analysis result
The percentage absorbance of the calculating of percentage absorbance, standard items or sample is equal to the absorbance of the mean value (diplopore) divided by first standard (0 standard) of the absorbance of standard items or sample, then is multiplied by 100%, i.e.,
Percentage absorbance (%)=B/B0×100%
Wherein mean absorbance values of B-standard liquid or sample solution, B0—0 The mean absorbance values of ppb standard liquids.
As abscissa, standard items percentage absorbance is ordinate to logarithm with the standard concentration (ppb) of Methomyl, draws calibration curve, obtains linear equation.The percentage absorbance of sample is substituted in calibration curve, the concentration corresponding to sample is read from calibration curve, be multiplied by the actual concentrations that its corresponding extension rate is Methomyl in sample.

Claims (7)

1. the enzyme linked immunological kit and its detection method of Methomyl, including ELISA Plate, Methomyl standard items, Methomyl antibody working solution, Methomyl ELIAS secondary antibody working solution, substrate solution A, substrate solution B, terminate liquid, concentration and dilution liquid and concentrated cleaning solution are detected.
2. the enzyme linked immunological kit and its detection method of Methomyl are detected, is comprised the following steps:Preparation, the preparation of Methomyl standard items, the preparation of Methomyl antibody working solution, the preparation of Methomyl ELIAS secondary antibody working solution, the preparation of substrate solution A, the preparation of substrate solution B, the preparation of terminate liquid, the preparation of concentration and dilution liquid and the preparation of concentrated cleaning solution of ELISA Plate.
3. it is according to claim 2 detection Methomyl enzyme linked immunological kit and its detection method, it is characterised in that:Described ELISA Plate preparation method is to be coupled Methomyl haptens and the pure albumen of carrier proteins Bovine (BSA) to obtain Methomyl envelope antigen, with carbonate (CBS) buffer solution of 0.05 mol/L pH 9.6 as coating buffer, envelope antigen is diluted to into 1:40000 ratios, 100 μ L/ holes, 37 DEG C of 2 h of incubation, take out ELISA Plate and get rid of liquid in plate, add the μ L/ holes of concentrated cleaning solution 300 after dilution, board-washing 2 times, 30 s/ time, it is subsequently adding 0.5% bovine serum albumin(BSA) (BSA) closing, 150 μ L/ holes, 37 DEG C of 1.5 h of placement, discard confining liquid and directly pat dry, ELISA Plate after patting dry is placed (25 DEG C) between constant temperature and is dried, and preserves under the conditions of ELISA Plate is vacuum-sealed in into 4 DEG C after sampling observation is qualified.
4. it is according to claim 2 detection Methomyl enzyme linked immunological kit and its detection method, it is characterised in that:The concentration of Methomyl standard items be respectively 0 mg/kg, 0.1mg/kg, 0.3 mg/kg、0.9 mg/kg、2.7 mg/kg、8.1 mg/kg。
5. it is according to claim 2 detection Methomyl enzyme linked immunological kit and its detection method, it is characterised in that:Described Methomyl antibody working solution is to obtain monoclonal antibody using Methomyl artificial antigen immune mouse, and with antibody diluent 1 is diluted to:It is prepared by 60000 ratios.
6. it is according to claim 2 it is a kind of detection Methomyl enzyme linked immunological kit and its detection method, it is characterised in that:Described Methomyl ELIAS secondary antibody working solution adds diluted into 1 by ELIAS secondary antibody:2000 ratios, the substrate solution A is the citrate-phosphate disodium hydrogen cushioning liquid of the carbamide peroxide containing 0.5 mmol/L, the substrate solution B is the ethanol solution of tetramethyl biphenyl diamines, the terminate liquid is the sulfuric acid of 2 mol/L, and the concentration and dilution liquid is 10 times of concentration and dilution liquid, and it is the PBS of 0.1 mol/L, between pH value range 7.0-7.5, the concentrated cleaning solution is 10 times of concentrated cleaning solutions, and it is containing 0.5% Tween-20 0.01 The PBST of mol/L, between pH value range 7.0-7.5.
7. the enzyme linked immunological kit and its detection method of the detection Methomyl described in claim 2, based on the indirect competitive enzyme-linked immunosorbent reaction principle of antigen-antibody, the method is characterized in that:Pretreatment testing sample,Take the ELISA Plate for being coated with Methomyl antigen,Sequentially it is separately added into standard items/sample、Methomyl ELIAS secondary antibody working solution、The each 50 μ L/ holes of Methomyl antibody working solution are in corresponding micropore,Gently vibration is mixed,30 min are reacted with cover plate membrane cover plate 25 DEG C of light protected environments of rearmounted room temperature,Liquid in hole is dried,4 ~ 5 times are fully washed with wash operating solution,Per the s of minor tick 10,The μ L/ holes of substrate solution A 50 are added after patting dry,The μ L/ holes of substrate solution B 50,Gently vibration is mixed,With 15~20 min of reaction in the rearmounted 25 DEG C of light protected environments of cover plate membrane cover plate,Add the μ L/ holes of terminate liquid 50,Gently vibration is mixed,Setting ELIASA is at 450 nm or the nm of dual wavelength 450/630 is detected,Determine per hole absorbance(Please run through data in 5 min), with standard concentration (ppb) logarithm as abscissa, standard items percentage absorbance be ordinate, draw calibration curve, reference standard curve calculate sample in Methomyl content.
CN201510694322.5A 2015-10-23 2015-10-23 Enzyme-linked immunosorbent assay kit for detecting methomyl and detection method thereof Withdrawn CN106610432A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201510694322.5A CN106610432A (en) 2015-10-23 2015-10-23 Enzyme-linked immunosorbent assay kit for detecting methomyl and detection method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201510694322.5A CN106610432A (en) 2015-10-23 2015-10-23 Enzyme-linked immunosorbent assay kit for detecting methomyl and detection method thereof

Publications (1)

Publication Number Publication Date
CN106610432A true CN106610432A (en) 2017-05-03

Family

ID=58612223

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201510694322.5A Withdrawn CN106610432A (en) 2015-10-23 2015-10-23 Enzyme-linked immunosorbent assay kit for detecting methomyl and detection method thereof

Country Status (1)

Country Link
CN (1) CN106610432A (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109682965A (en) * 2019-02-01 2019-04-26 湘潭大学 Microwell plate envelope antigen/antibody method is realized with flat plate ink-jet printer
CN115073337A (en) * 2022-05-16 2022-09-20 江南大学 Methomyl hapten, holoantigen, monoclonal antibody, preparation method and application thereof

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103808934A (en) * 2012-11-07 2014-05-21 江苏维赛科技生物发展有限公司 Preparation of ELISA kit for detecting deoxynivalenol
CN104655846A (en) * 2013-11-20 2015-05-27 镇江先创生物科技有限公司 Enzyme linked immunosorbent assay kit for detecting progesterone and detection method thereof
CN104655847A (en) * 2013-11-20 2015-05-27 南京亿特生物科技有限公司 Enzyme linked immunosorbent assay kit (ELISA kit) for detecting adprin and detection method thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103808934A (en) * 2012-11-07 2014-05-21 江苏维赛科技生物发展有限公司 Preparation of ELISA kit for detecting deoxynivalenol
CN104655846A (en) * 2013-11-20 2015-05-27 镇江先创生物科技有限公司 Enzyme linked immunosorbent assay kit for detecting progesterone and detection method thereof
CN104655847A (en) * 2013-11-20 2015-05-27 南京亿特生物科技有限公司 Enzyme linked immunosorbent assay kit (ELISA kit) for detecting adprin and detection method thereof

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109682965A (en) * 2019-02-01 2019-04-26 湘潭大学 Microwell plate envelope antigen/antibody method is realized with flat plate ink-jet printer
CN109682965B (en) * 2019-02-01 2022-02-18 湘潭大学 Method for realizing antigen/antibody coating of microporous plate by flat ink-jet printer
CN115073337A (en) * 2022-05-16 2022-09-20 江南大学 Methomyl hapten, holoantigen, monoclonal antibody, preparation method and application thereof

Similar Documents

Publication Publication Date Title
CN104655846A (en) Enzyme linked immunosorbent assay kit for detecting progesterone and detection method thereof
CN104655847A (en) Enzyme linked immunosorbent assay kit (ELISA kit) for detecting adprin and detection method thereof
CN105510589A (en) Enzyme linked immunosorbent assay (ELISA) kit for detecting carbendazim and detection method thereof
CN105319368A (en) Enzyme linked immunosorbent assay kit used for detecting zearalenone, and detection method thereof
CN101650368A (en) Method for testing zearalenone toxin by using colloidal gold immunochromatography assay
CN105572336A (en) Enzyme linked immunosorbent assay kit for detecting imidacloprid and detecting method of kit
CN106568961A (en) Enzyme linked immunosorbent assay kit for detection of paraquat and detection method thereof
CN106610432A (en) Enzyme-linked immunosorbent assay kit for detecting methomyl and detection method thereof
CN106610431A (en) Enzyme-linked immunosorbent assay kit for detecting Benalaxyl and detection method thereof
CN102331500A (en) Method and enzyme linked immunosorbent assay kit for detecting lemon yellow
CN105319353A (en) Preparation of enzyme linked immunosorbent assay kit used for detecting dihydropyridine drug residues
CN105301244B (en) Detect enzyme linked immunological kit and its application of acid orange
CN103102319A (en) Melamine semiantigen, preparation method and application thereof
CN107064494B (en) Enzyme linked immunosorbent assay kit for chlorpyrifos detection
CN115541882A (en) Preparation method and kit for improving detection test strip of human immunodeficiency virus antibody
CN105510588A (en) Enzyme-linked immunoassay (ELISA) kit for detecting glyphosate and detection method thereof
CN111289753B (en) Enzyme linked immunosorbent assay kit for detecting coumarin and indandione rodenticide and preparation and application thereof
CN111443202B (en) ELISA kit for detecting anticoagulant rodenticide, preparation and application thereof
CN106610426A (en) Enzyme linked immunosorbent assay kit used for detecting acetochlor, and detection method thereof
CN105301245A (en) Preparation method of enzyme-linked immunosorbent assay (ELISA) kit for detecting cyproheptadine hydrochloride
CN111879936A (en) Enzyme linked immunosorbent assay kit for detecting vomitoxin in edible oil and detection method thereof
CN106568964A (en) Enzyme linked immunosorbent assay kit for detecting butachlor and detection method thereof
CN113125728A (en) Preparation of enzyme linked immunosorbent assay kit for detecting variegated aflatoxin
CN108226476A (en) A kind of enzyme linked immunological kit and its detection method for detecting Madumycin
CN105277705A (en) Preparation method and detection method for biguanide drug residue detection kit

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
WW01 Invention patent application withdrawn after publication

Application publication date: 20170503

WW01 Invention patent application withdrawn after publication