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ELISA kit for detecting carbofuran

An enzyme-linked immunosorbent reagent, the technology of carbovir, which is used in biological testing, measuring devices, material inspection products, etc., can solve the problems of low degree of instrumentation, high cost, expensive professional instruments, etc., to achieve simple operation, saving consumption, The effect of increasing sensitivity

Inactive Publication Date: 2007-08-01
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the conventional methods for detecting carbofuran residues include gas chromatography (GC) and high performance liquid chromatography (HPLC). Although these methods are sensitive and accurate, they require expensive professional instruments, and the analysis is time-consuming and costly.
The enzyme-linked immunoassay (ELISA) rapid detection technology has become a commonly used screening method due to its low cost, simple operation, fast speed, large sample size for one test, and low degree of instrumentation, but there is no carbofuran immunoassay reagent at present. emergence of box

Method used

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  • ELISA kit for detecting carbofuran
  • ELISA kit for detecting carbofuran
  • ELISA kit for detecting carbofuran

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The preparation of embodiment 1 gram Budweiser ELISA kit sample

[0018] 1. Buffer preparation buffer (pH7.4PBST) KH 2 PO 4 0.4g, Na 2 HPO 4 12H 2 O 5.8g, NaCl 16g, KCl 0.4g, Tween-20 0.05% 1mL, add distilled water to 2000mL.

[0019] 2. Preparation of blocking solution Dissolve 1.0-5.0 g of skimmed milk powder in 100 mL of distilled water.

[0020] 3. Preparation of substrate solution Dissolve 30 μL of 30% hydrogen peroxide in 19 mL of chromogenic solution (pH 5.0 phosphoric acid-citric acid buffer 0.2 M Na 2 HPO 4 25.7mL, 0.1M citric acid 24.3mL, add distilled water 50mL), store at 4°C.

[0021] 4. Preparation of substrate buffer solution Dissolve 80 mg o-phenylenediamine OPD in 10 mL substrate solution and store at 4°C.

[0022] 5. Dilute the second antibody coated on the microplate with pH 9.6, 0.05mol / L carbonate buffer solution (containing 1-2g sodium carbonate and 2-4g sodium bicarbonate, 1L double distilled water) to 0.5- 5ug / mL, add 100uL to each wel...

Embodiment 2

[0033] Embodiment 2 detects the pretreatment of sample:

[0034] Water sample: After filtration, samples can be taken for ELISA analysis.

[0035]Soil sample: Take 10g of soil and extract it three times with 20-40mL of methanol, combine the extracts, concentrate, then dilute with PBST to 10mL for ELISA analysis.

[0036] Vegetable samples: Take vegetable samples and crush them with a pulverizer, weigh 10g, extract three times with 20-40mL of methanol, combine the extracts, concentrate, dilute to 10mL with PBST, and take samples for ELISA analysis.

[0037] Blood: take human blood, add anticoagulant and directly analyze it by ELISA method.

[0038] Gastric lavage solution (2% sodium bicarbonate solution): take 10 mL of gastric lavage solution, adjust the pH value to neutral with dilute HCl, and then analyze it by ELISA.

[0039] Vomitus: Take the sample and grind it, centrifuge to get the supernatant and analyze it by ELISA method.

Embodiment 3

[0040] Embodiment 3 detection method

[0041] The operation process of the kit is as follows: take out an ELISA plate coated with the secondary antibody, and return it to room temperature for later use; add 100uL of carbofuran antibody to each well, incubate at 37°C for 0.5h, wash the plate, and add 50uL of a series of concentrations Carbofuran standard sample and 50uL enzyme-labeled antigen were used for competition reaction, incubated at 37°C for 1h, washed, OPD was developed at 37°C in the dark for 15min, terminated with sulfuric acid, and the results were measured with a microplate reader at 490nm.

[0042] Calculate the inhibition rate based on the average value of the sample absorbance values ​​obtained, and the calculation formula for the inhibition rate is:

[0043]

[0044] Among them, Amax is the absorbance value when no pesticide is added, Ax is the absorbance value when the pesticide concentration is x, and Amin is the absorbance value of the blank control well....

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Abstract

This invention discloses one enzyme immune agent base to test carbofuran, which comprises base body and enzyme label board, which is characterized by the following: anti-carbofuran antigen, horse radish hyperoxide enzyme label antigen, carbofuran standard liquid, bottom liquid, bottom buffer liquid and terminal liquid, wherein, the said enzyme standard board covers anti-carbofuran antigen abnormal second antigen. This invention enzyme immune agent case uses second antigen covering enzyme standard board.

Description

technical field [0001] The invention relates to the field of ELISA detection, in particular to an ELISA kit for detecting carbofuran. Background technique [0002] Carbofuran (carbofuran, chemical name 2,3 dihydro-2,2 dimethyl-7-benzofuryl-N-methylcarbamate), trade name carbofuran, has been produced by FMC and After being developed and produced by Mobay Chemical Company, it is used as a high-efficiency and broad-spectrum insecticide, which is widely used in the control of pests in grains, vegetables, fruits and economic crops. Carbofuran is a cholinesterase inhibitor that is highly toxic to humans and wild animals (oral LD ​​in mice 50 8mg / kg), it is a kind that cannot be detected in the regulations on pesticide residues in vegetables, but due to its good insecticidal effect, the phenomenon of unreasonable use in food and vegetables is relatively serious, and it has great harm to public health . In addition, carbofuran is not easy to degrade in acidic soil, and it is easy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/547G01N33/577G01N21/78G01N33/53
Inventor 孙远明杨金易潘科王弘吴青雷红涛肖治理谌国莲沈玉栋
Owner SOUTH CHINA AGRI UNIV
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