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Chemiluminescence-based biochip capable of simultaneously detecting various pesticide and veterinary drug residues and detecting method

A biochip and chemiluminescence technology, applied in the field of immunoassay, can solve the problem that the kit is difficult to meet the actual detection, and achieve the effect of short detection time, high sensitivity, and simple operation

Inactive Publication Date: 2019-04-12
NANJING XIANGZHONG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the existing research at home and abroad is mainly focused on the establishment of specific immunoassay methods for a single agricultural and veterinary drug. In fact, there are often multiple agricultural and veterinary drug residues in agricultural products at the same time. A single agricultural and veterinary drug immunoassay method and corresponding kits It has been difficult to meet the needs of actual detection

Method used

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  • Chemiluminescence-based biochip capable of simultaneously detecting various pesticide and veterinary drug residues and detecting method
  • Chemiluminescence-based biochip capable of simultaneously detecting various pesticide and veterinary drug residues and detecting method
  • Chemiluminescence-based biochip capable of simultaneously detecting various pesticide and veterinary drug residues and detecting method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The contents of tetracycline, streptomycin, sulfamethoxazole, ciprofloxacin, carbendazim and cyfluthrin in honey samples were also detected.

[0044] The reagents used in the experiment are as follows:

[0045] (1) Standard solution:

[0046] There are 6 kinds of mixed standard solutions used in the present embodiment, and the composition is as follows:

[0047] ①Mixed standard solution 1: tetracycline 0ng / mL, streptomycin 0ng / mL, sulfamethoxazole 0ng / mL, ciprofloxacin 0ng / mL, carbendazim 0ng / mL, and cyfluthrin 0ng / mL.

[0048] ②Mixed standard solution 2: tetracycline 0.2ng / mL, streptomycin 0.15ng / mL, sulfamethoxazole 1ng / mL, ciprofloxacin 0.05ng / mL, carbendazim 0.1ng / mL, cyfluthrin 2ng / mL.

[0049] ③Mixed standard solution 3: tetracycline 0.6ng / mL, streptomycin 0.45ng / mL, sulfamethoxazole 3ng / mL, ciprofloxacin 0.15ng / mL, carbendazim 0.3ng / mL, cyfluthrin 6ng / mL.

[0050] ④Mixed standard solution 4: tetracycline 1.8ng / mL, streptomycin 1.35ng / mL, sulfamethoxazole 9ng...

Embodiment 2

[0080] At the same time, the contents of clenbuterol, ractopamine, sulfadiazine, doxycycline, ofloxacin, florfenicol, lincomycin, and phenylethanolamine A in pork samples were quantitatively detected.

[0081] The detection steps are as follows:

[0082] (1) Preparation of biochip: adopt the same method as Example 1, the difference is: use clenbuterol-BSA, ractopamine-BSA, sulfadiazine-KLH, doxycycline-OVA, ofloxacin-BSA , florfenicol-BSA, lincomycin-OVA, phenylethanolamine A-BSA replace the antigen in embodiment 1;

[0083] (2) Preparation of the sample solution to be tested: take 5 g of crushed pork sample, add 8 mL of buffer, mix for 5 min, place in a water bath at 50°C for 30 min, centrifuge for 10 min, take 50 μL of supernatant, add 450 μL of buffer and mix, take 50 μL The sample solution to be tested is obtained;

[0084] (3) Add 25 μL of free clenbuterol, ractopamine, sulfadiazine, doxycycline, ofloxacin, florfenicol, and lincomycin with different concentration gradie...

Embodiment 3

[0088] At the same time, the residues of alachlor, chlorothalonil, deltamethrin, imidacloprid, methyl parathion, carbendazim, carbofuran and carbaryl in Chinese cabbage were quantitatively detected.

[0089] (1) Preparation of biochip: adopt the same method as Example 1, the difference is: use alachlor-OVA, chlorothalonil-BSA, deltamethrin-BSA, imidacloprid-BSA, methyl parathion- BSA, carbendazim-BSA, carbofuran-BSA, carbaryl-BSA replace the antigen in Example 1;

[0090] (2) Preparation of the sample solution to be tested: Take 5g of cabbage leaves, add 5mL of water, crush them, add 10mL of ethyl acetate, vortex and centrifuge; take 5mL of the centrifuged upper layer solution, blow dry under nitrogen flow; dry and add 1mL of PBS solution, shake to dissolve, and the obtained solution is ready for use;

[0091] (3) Add 50 μL of free alachlor, chlorothalonil, deltamethrin, imidacloprid, methyl parathion, carbendazim, carbofuran, For the mixed standard solution of carbaryl, add...

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Abstract

The invention provides a chemiluminescence-based biochip capable of simultaneously detecting various pesticide and veterinary drug residues. The biochip comprises a chip carrier where a set of detecting target antigens is fixed, detecting targets are pesticides and / or veterinary drugs, and the biochip is manufactured through a method comprising the following steps that with bovine serum albumin asblank control, the bovine serum albumin and the detecting target antigens are subjected to point-sampling operation on the chip carrier through a biochip manufacturing system, then incubation in a water bath of 20-37 DEG C is carried out for 0.5-4 h, and the biochip can be obtained after washing and air-drying. The invention further provides a method for detecting various pesticides and veterinary drugs through chemiluminescence and the biochip. The biochip is simple in structure and manufacturing process, low in cost, large in target quantity, high in accuracy, sensitivity and precision, short in detection time and easy and practicable to operate, no expensive detecting instrument is needed, and the biochip is suitable for primary screening of a large scale of samples on site.

Description

technical field [0001] The invention belongs to the immunological analysis technology of food safety supervision or food analysis, in particular to a biochip and detection method based on chemiluminescence that can simultaneously detect residues of various agricultural and veterinary drugs. Background technique [0002] The use of pesticides and veterinary drugs is currently an effective means for countries all over the world to prevent crop diseases and insect pests in agricultural planting and prevent diseases in livestock and poultry aquaculture. However, the use of a large number of pesticides and veterinary drugs has brought a huge security threat to the entire ecosystem and food safety, and has become a "killer" that endangers human health. [0003] Pesticide residues are the general term for the trace pesticide precursors, toxic metabolites, degradation products and impurities that remain in organisms, crops, soil, water, and the atmosphere without being decomposed wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/02
CPCG01N33/02
Inventor 李钟卉李周敏
Owner NANJING XIANGZHONG BIOTECH
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