Pesticide and veterinary drug multi-residue detection method based on microarray detection chip

Abstract

The invention relates to a pesticide and veterinary drug multi-residue detection method based on a microarray detection chip. The method comprises the steps that a micromolecular pesticide and veterinary drug hapten is coupled with bovine serum albumin to prepare an immunizing antigen; a mouse is immunized by the immunizing antigen to prepare a corresponding pesticide and veterinary drug monoclonal antibody; a biochip sample application instrument for a monoclonal antibody acquisition probe performs sample application on an agarose modified slide solid phase carrier to prepare a multi-repeat monoclonal antibody acquisition probe microarray; the pesticide and veterinary drug hapten is coupled with ovalbumin to prepare a hapten-OVA (ovalbumin) couplet; a fluorescent molecule Cy3 is then used for labeling; sample liquid to be detected is incubated with the chip after mixed with a labeling detection antigen according to a certain concentration; a to-be-detected antigen in a sample and the corresponding labeling detection antigen are directly and competitively bound with the corresponding monoclonal antibody acquisition probe fixed on the chip; elution is performed under a certain condition; and a result is detected by a biochip scanner. The method can detect multiple pesticide and veterinary drug residues in the subsidiary agricultural product sample simultaneously, and is applicable to high-throughput, quick and accurate detection of drug residues in planting and culture production.

Description

[0001] technical field [0002] The invention relates to a method for preparing a multi-residue detection microarray chip of agricultural and veterinary drugs in the field of food safety. [0003] Background technique [0004] Agricultural and veterinary drugs are effective means to prevent and control crop diseases and insect pests and aquaculture diseases in countries around the world. The use of a large number of chemical pesticides and veterinary drugs, especially the highly toxic and high-residue pesticides and veterinary drugs, has become the main "killer" of food safety and endangering people's health. [0005] At present, the main chemical pesticides used are mainly divided into three categories: insecticides, herbicides and fungicides according to their different functions. According to different chemical structures, they can be divided into organochlorines, organophosphorus, pyrethroids, carbamates, and inorganic pesticides. The common characteristics of the...

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Problems solved by technology

(3) Chromatography-mass spectrometry (GC-MS, HPLC-MS): It can achieve qualitative and quantitative detection purposes at the same time, especially suitable for the detection of pesticide metabolites, degradation products and multi-residue detection, etc., but this method requires expensive instruments And the operation is complicated and difficult, not suitable for frequent detection

(4) Supercritical Fluid Chromatography (SFC): It not only has the characteristics of fast, efficient, and sensitive gas spectroscopy, but also has the characteristics of liquid chromatography that can detect thermally unstable and macromolecular compounds, but requires expensive instruments and complicated operations Difficulty, etc.

(5) Capillary electrophoresis (CE): It is suitable for the separation and analysis of some ionized samples that are difficult to separate by traditional chromatography. It has 10-1000 times higher analytical capacity than HPLC. It requires expensive instruments and is complicated and difficult to operate. It is not suitable for for regular testing

However, due to the influence of antibodies such as dependence on foreign imports, at present, the ELISA finished kits in the Chinese market rely on imports from abroad.

(3) Enzyme inhibition method is the most mature and widely used rapid pesticide residue detection technology. This method can only detect organophosphorus and carbamate pesticides, and the inhibition rate of different pesticides of the same kind varies greatly. , so using a uniform inhibition rate to determine whether pesticide residues exceed the standard will inevitably lead to false positive or false negative missed detection, which can be said to be the current "rapid detection method adopted as a last resort", which is only applicable to the initial inspection at the grassroots level and plays a warning role , when the phenomenon of exceeding the standard is found, it must be retested and confirmed by standard methods, negative reactions should also be sampled in proportion, and retested and confirmed by reliable methods

[0015] Existing research at home and abroad is mainly focused on establishing a specific immunoassay method for a single pesticide and veterinary drug, but in fact there are often many kinds of pesticide and veterinary drug residues in agricultural products, Immunoassay methods and corresponding detection kits for single agricultural and veterinary drug components have been difficult to meet the needs of actual detection

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