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Pesticide fenthion immunity detecting method

An immunological detection method, fenthion technology, applied in the biological field, can solve the problems of high detection cost, low detection efficiency, complicated operation, etc., and achieve high accuracy, good practicability, and good repeatability

Inactive Publication Date: 2007-09-19
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Technical problem The object of the present invention is to provide the fenthion ELISA detection method for the fenthion detection method in the existing chromatographic instrument technology, which has complicated operation, high detection cost and low detection efficiency, so as to find out the fenthion pesticide residue. A fast, accurate, simple and low-cost ultra-trace detection method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Detecting fenthion residues in the river water sample of embodiment 1

[0044] Preparation of fenthion polyclonal antibody

[0045] Using the immunogen "fenthion-bovine serum albumin conjugate" of fenthion artificial antigen, New Zealand white rabbits were immunized with conventional immunization methods, and the rabbit antiserum against fenthion was prepared, which was fenthion polyclonal antibody. The specific operation method refers to the research paper "Establishment and Preliminary Application of Indirect Competitive ELISA for Quantitative Determination of Methamidophos Residues" by Liu Fengquan et al., which was published in "Journal of Agricultural Biotechnology", Vol. Page.

[0046] Main conditions of fenthion ELISA reaction system

[0047] "Fenthion-Ovalbumin Conjugate" coating 1.2μg / ml, add 50μl to each well of the microanalysis plate, dilute the fenthion antiserum 16000 times as the working concentration, 1% gelatin as the block, antigen antibody The meth...

Embodiment 2

[0072] Example 2 Detection of Fenthion Residues in Peach Fruit Samples

[0073] The main conditions of the fenthion ELISA reaction system are the same as in Example 1.

[0074] Sample preparation for testing

[0075] Peach fruit sample: Weigh 100 g of chopped peach fruit, add fenthion with a net weight of 2 μg, and prepare a solid sample of 20 ppb.

[0076] The above samples were detected by the fenthion ELISA detection method, and the operation was as follows:

[0077] Steps (1), (2) are the same as in Example 1.

[0078] (3) Add antibody and the mixture of antibody and sample to be tested

[0079] Weigh 0.1g of the sample to be tested, soak it in 0.2ml methanol for about 1 hour, take 100μL and add it to 900μL reaction matrix to prepare the test solution, dilute 8000 times fenthion antiserum with the test solution and blank matrix solution in equal volume Mix well, then add 50 μL per well into the micro-analysis plate, react at 37°C for 1 hour, pour it out, wash with regu...

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Abstract

The present invention relates to an immunoassay for detecting pesticide fenthion residues and belongs to the field of biological technology. In an immunoassay for detecting pesticide fenthion residues ELISA, 'fenthion-bovine serum albumin conjugate' is adopted as a immunogen prepared fenthion antiserum (namely fenthion polyclonal antibody) with a detection equation of Logit(B / Bo)=-0.7575LogC-1.4928 (wherein B is a sample hole absorption value, Bo is a positive contrast absorption value, and C represents the fenthion cnocentration measured from a board hole), wherein the measurement conditions are as follows: 50 mu l fenthion coated antigen 'fenthion-ovalbumin conjugate' of 1.2 mu g / ml is added into every hole of a trace analysis plate, the fenthion antiserum is diluted to 16000 times to operate as a working concentration, 1% glutin is adopted as a sealing material, an antigen antibody reaction solution of phosphate-tween buffer solution contains methanol of 5% and has a pH of 7.4 and an ion intensity of 0.15 mol / L. Said method has remarkable specificity, stability and sensitivity, and a technic method used for detecting fenthion quickly, sensitively and correctly is provided.

Description

(1) Technical field [0001] The invention relates to an enzyme-linked immunosorbent assay (ELISA) detection method for pesticide fenthion residue, and belongs to the field of biotechnology. It is dedicated to the high-sensitivity and rapid detection of fenthion pesticide residues in agricultural products and agricultural production environments, and is especially suitable for large-scale sample detection and on-site monitoring. (2) Background technology [0002] Fenthion is an organophosphorus insecticide with a structure of It has moderate toxicity, wide insecticidal spectrum, long residual effect period and is widely used. Fenthion residues are harmful to humans and livestock, and the widespread use of fenthion will inevitably bring harm to environmental sanitation and agricultural product safety. Therefore, all countries in the world have made strict regulations on the maximum allowable residue of fenthion in agricultural products, and my country is no exception. [00...

Claims

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Application Information

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IPC IPC(8): G01N33/543
Inventor 刘凤权张奇胡白石
Owner NANJING AGRICULTURAL UNIVERSITY
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