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Method for preparing and using reagent for simultaneously detecting multiple small molecular compounds

A small molecular compound, simultaneous detection technology, applied in the direction of measuring devices, analytical materials, instruments, etc., can solve the problems of complex operation steps, long detection cycle, inability to conduct on-site, rapid detection, etc.

Inactive Publication Date: 2007-04-11
邹明强
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, small molecular compounds that are only antigenic but not immunogenic, such as pesticides, veterinary drugs, food additives, mycotoxins, and illegal drugs, have only one site that can bind to antibodies, and double-antibody sandwich immunity cannot occur reaction
Therefore, the flow cytometry, suspension chip technology or liquid phase chip technology of the above-mentioned encoded microspheres cannot be used for the detection of small molecular compounds that are only antigenic but not immunogenic
At present, the common methods used in the detection of small molecular compounds, such as pesticides, veterinary drugs, food additives, mycotoxins, prohibited drugs and other hazards, such as GC, HPLC, TLC, CZE, MS, MS-MS, etc., there are instruments The price is expensive, the operation steps are complicated, the detection cycle is long, it is difficult to popularize and promote, and it is not possible to conduct on-site and rapid detection, which is not conducive to timely discovery and monitoring of management problems

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Preparation of reagents required for a kit for simultaneous detection of multiple small molecule compounds

[0059] 1. Prepare the coded microsphere reagent according to the above method, in the third step, take 1μL each of the polystyrene fluorescent microspheres with a particle diameter of 3μm, a fluorescence intensity of level 2, and a fluorescence color of red, orange, yellow or green; or The particle diameter is 10μm, the fluorescence color is green, the fluorescence intensity is 1, 2, 4, 5, 6 or 8 polystyrene fluorescent microspheres each 5μL; or the fluorescence color is red, the fluorescence intensity is 3, the particle diameter 3μL each of 4, 5, 6, 8, 9μm polystyrene fluorescent microspheres; among them, in step 5, make the weight percentage concentration of BSA be 2%, 5% or 10%; among them, in step 6, use 100μL each time , 150μL or 200μL blocking solution wash 2 times.

[0060] 2. Prepare fluorescent quantum dot antibody probes according to the above met...

Embodiment 2

[0064] Example 2 Assembly of a kit for simultaneous detection of multiple small molecule compounds (100 tests can be performed)

[0065] 1. One or more different small molecule compound coding microsphere reagents:

[0066] A kit for the detection of a certain small molecule compound, with this small molecule compound coded microsphere reagent 100 times the required amount of 10mL, such as malachite green detection kit with malachite green coded microsphere reagent 10mL, or more All reagents are stored at 4°C.

[0067] A kit for a variety of small-molecule compound detection tests, each with 10 mL of each small molecule compound-encoded microsphere reagent for 100 experiments, and at least the particle size, fluorescence color or fluorescence between the coded microspheres in the kit There is a difference in one of the intensities, such as simultaneous detection of clenbuterol hydrochloride, ractopamine, and chloramphenicol, with clenbuterol hydrochloride with a particle size of 3...

Embodiment 3

[0080] How to use the kit for simultaneous detection of multiple small molecule compounds-antibody probe direct method to detect a single small molecule compound, for example, to detect the content of clenbuterol in a sample:

[0081] a. Drawing the standard curve in the kit

[0082] 1) Take 100 μL of the standard mass concentration solution of Clenbuterol Hydrochloride Antigen Series with a mass concentration of 0ng / ml, 0.1ng / ml, 0.3ng / ml, 0.9ng / ml, 2.7ng / ml and 8.1ng / ml respectively in a centrifuge tube Add 100μL of coded microsphere reagent prepared by immobilizing clenbuterol whole antigen with activated microsphere solution with a particle size of 4μm, red fluorescence, and fluorescence intensity of level 3, and then add 10μL of clenbuterol hydrochloride. Clone the antibody green fluorescent quantum dot probe, and shake it for 0.5 hours in the dark to cause a competitive immune reaction to generate a coded microsphere-fluorescent quantum dot antibody probe complex;

[0083] 2...

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PUM

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Abstract

The present invention provides one kind of reagent for simultaneously detecting several kinds of small molecular compounds and its preparation process and usage. The preparation process includes the following steps: preparing coded microsphere reagent, and preparing one of the small molecular compound detecting antibody probes, including fluorescent quantum dot antibody probes, fluorescent quantum dot secondary antibody probes, fluorescent dye antibody probes and fluorescent dye secondary antibody probes. The method of detecting small molecular compounds with the antibody probe may be a direct antibody probe method or a indirect antibody probe method. The present invention may be used in detecting residual veterinary medicine, pesticide, banned drug, misused medicine and other small molecular compounds in food, agricultural product, living animal body and human body, and has the advantages of high sensitivity, high repeatability, etc.

Description

Technical field [0001] The present invention relates to a method for preparing reagents, in particular to a method for preparing reagents for simultaneous detection of multiple small molecule compounds, and also to a method for using the reagents. Background technique [0002] Residues of veterinary drugs, pesticides, food additives, and mycotoxins seriously threaten people's health and life safety. Chloramphenicol, clenbuterol, diethylstilbestrol, nitrofurans and malachite green are the key monitoring targets. In addition, illegal drugs and abused drugs such as morphine are extremely harmful. The timely, accurate, and convenient detection of these hazards is one of the keys to protecting human health and life safety, and maintaining social stability. [0003] The above-mentioned hazards are all small molecule compounds, and conventional detection methods are mainly divided into two types: physical and chemical analysis methods and immunological analysis methods. The physical and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N33/577G01N33/546
Inventor 邹明强李锦丰高海霞
Owner 邹明强
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