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Fast food safety homogeneous immunological detection reagent

An immunodetection and reagent technology, applied in the field of food safety detection reagents, can solve problems such as low sensitivity, and achieve the effects of high sensitivity, rapid detection, and improved detection accuracy and efficiency.

Inactive Publication Date: 2010-12-29
CHANGZHOU SEO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Commercially available colloidal gold test paper can only be used for qualitative detection. The detection results of ELISA kits are affected by the environment and operator skills. The sensitivity of traditional EMIT kits is low.

Method used

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  • Fast food safety homogeneous immunological detection reagent
  • Fast food safety homogeneous immunological detection reagent
  • Fast food safety homogeneous immunological detection reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Ractopamine Homogeneous ELISA Technology Product

[0029] 1 Preparation of hapten

[0030] Dissolve 0.8 g of ractopamine and 0.3 g of acetic anhydride in 20 ml of tetrahydrofuran, add 0.5 ml of diethylamine, and heat to reflux for 12 hours. After removing the solvent under reduced pressure, 10 ml of hydrochloric acid solution (0.1 N) was added, and the target hapten was extracted with 30 ml of ethyl acetate. After removal of ethyl acetate, the resulting semi-solid (0.75 g) was dissolved in 10 ml of methanol. It is directly used for the synthesis of the complete antigen in the next step.

[0031] 2 Antigen synthesis

[0032] Take 1ml of the above methanol solution (75mg hapten), remove the solvent under reduced pressure, dissolve the residue in 0.5ml of diformamide (DMF), add 0.1g of DCC and 0.03g of nitrogen hydroxysuccinamide, and react at room temperature for 2 hours, The reaction was added in portions to 5 ml of protein such as BSA (0.3 g) or BTG, KLH ...

Embodiment 2

[0046] Example 2: Clenbuterol hydrochloride homogeneous ELISA product

[0047] 1 Preparation of haptens and whole antigens

[0048] The method is the same as the preparation of the hapten in Example 1 and the synthesis of the antigen in Example 1.

[0049] 2 Antibody preparation

[0050] Inoculate experimental animals (rabbits) with the obtained complete antigen according to conventional methods, and obtain antiserum after booster immunization.

[0051] 3 Preparation of enzyme-labeled antigen

[0052] The method is the same as the preparation of the enzyme-labeled antigen in Example 1.

[0053] 4 Preparation of homogeneous enzyme-linked reagents

[0054] The method is the same as the preparation of the homogeneous enzyme-linked reagent in Example 1.

[0055] 5 Determination of clenbuterol content in pig urine

[0056] The operation process is the same as in Embodiment 1. The following data are measured by the automatic biochemical analyzer Oympus 400E:

[0057] Clenbut...

Embodiment 3

[0059] Embodiment 3: Benzodiazepine compound homogeneous ELISA product

[0060] This patent reports for the first time that the detection sensitivity of benzodiazepines can reach 1ng / ml, while the traditional method can only reach about 10ng / ml. This patent uses an artificially synthesized improved specific G6PDH enzyme, which successfully increases the detection sensitivity by about 10 times. The detection data are as follows (automatic biochemical analyzer Oympus 400E):

[0061] Benzodiazepine EMIT test result table

[0062] ng / ml

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Abstract

The invention relates to the field of biotechnology, in particular to a homogeneous immunological detection reagent. The invention adopts a method utilizing the homogeneous enzyme multiplied immunoreaction to detect the residual amounts of harmful substances in the body fluid of animal and the extract liquor of tissue, thus harmful substance residues containing common organic small-molecule medicines and illegal medicines, such as clenbuterol hydrochloride, ractopamine, melamine, malachite green, benzodiazepines, tricyclic antidepressants and various antibiotic medicines can be detected and the invention has extremely wide application prospect and social meaning. The product developed by the invention has high sensitivity and can detect hundreds of compounds and metabolic products at the same time; and the detection is fast and accurate; and the detection data can be processed and printed automatically and can also be connected with the Internet so as to realize synchronous monitoring.

Description

Technical field [0001] The invention relates to the field of biotechnology, in particular to a food safety detection reagent. Background technique [0002] The Ministry of Health and the Ministry of Agriculture of my country have formulated a series of pesticide residue limits (about 136 kinds) and corresponding 123 kinds of pesticide inspection methods, and promulgated the multi-residue detection methods of pesticides (including organochlorine, organophosphorus, pyrethroids and carbamates). nearly 50 kinds of pesticides). However, my country still lacks self-developed systematic multi-residue, high-throughput homogeneous ELISA technology products. [0003] Immunoassay method is currently the fastest way to detect small molecule chemical drugs. According to the survey conducted by the European Community on more than 2,000 laboratories (including forensic laboratories, clinical laboratories, research institutions, and reference laboratories), screening experiments use The im...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/531G01N33/535G01N21/31C07C211/52C07C215/60C07D251/54C07C251/30
Inventor 王国洪王志洪
Owner CHANGZHOU SEO BIOTECH
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