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

A technology for simultaneous detection of small molecular compounds, applied in measuring devices, analytical materials, instruments, etc., can solve problems such as inability to detect small molecular compounds, long detection cycle, and difficulty in popularization

Inactive Publication Date: 2011-06-08
邹明强
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] 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

[0059] Example 1 Preparation of reagents required for the kit for simultaneous detection of multiple small molecule compounds

[0060] 1. Prepare the coded microsphere reagent according to the above method, in the third step, take 1 μL of polystyrene fluorescent microspheres with a particle diameter of 3 μm, a fluorescence intensity of level 2, and a fluorescent color of red, orange, yellow or green; or take 5 μL each of polystyrene fluorescent microspheres with a particle diameter of 10 μm, a fluorescent color of green, and a fluorescence intensity of 1, 2, 4, 5, 6, or 8; or a fluorescent color of red, a fluorescence intensity of 3, and a particle diameter of 3 μL each of 4, 5, 6, 8, and 9 μm polystyrene fluorescent microspheres; in step 5, make the concentration of BSA by weight 2%, 5% or 10%; in step 6, use 100 μL each time , 150μL or 200μL blocking solution and washed twice.

[0061] 2. Prepare fluorescent quantum dot antibody probes as described above, wherein in the 3rd...

Embodiment 2

[0065] Example 2 Assembly of kits for simultaneous detection of multiple small molecule compounds (can be tested 100 times)

[0066] 1. One or more different small molecule compound-encoded microsphere reagents:

[0067] For a kit for the detection of a certain small molecule compound, the amount required for 100 experiments with this small molecule compound encoded microsphere reagent is 10mL, for example, the malachite green detection kit is equipped with 10mL malachite green encoded microsphere reagent, above All reagents were stored at 4°C.

[0068] Kits for various small molecular compound detection reagents, respectively equipped with 10mL of coded microsphere reagents required for 100 experiments of each small molecular compound, and the coded microspheres in the kit have at least particle size, fluorescent color or fluorescence There is a difference in one of the intensity, such as simultaneous detection of clenbuterol hydrochloride, ractopamine, and chloramphenicol, ...

Embodiment 3

[0081] The method of using the kit for simultaneous detection of multiple small molecule compounds - the antibody probe direct method to detect a single small molecule compound, for example, to detect the content of clenbuterol hydrochloride in a sample:

[0082] a. Drawing of the standard curve prepared in the kit

[0083] 1) Take 100 μL of clenbuterol hydrochloride antigen series standard mass concentration solutions with mass concentrations of 0ng / ml, 0.1ng / ml, 0.3ng / ml, 0.9ng / ml, 2.7ng / ml and 8.1ng / ml respectively in centrifuge tubes Add 100 μL of coded microsphere reagent prepared by immobilizing clenbuterol hydrochloride whole antigen with activated microsphere solution with a particle size of 4 μm, red fluorescence, and fluorescence intensity of grade 3, and then add 10 μL of clenbuterol hydrochloride single Clone the antibody green fluorescent quantum dot probe, shake it in the dark for 0.5 hours to make it undergo a competitive immune reaction, and generate a coded mi...

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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 invention relates to a preparation method of a reagent, in particular to a preparation method of a reagent for synchronously detecting multiple small molecule compounds, and also relates to a method for using the reagent. Background technique [0002] Residues of veterinary drugs, pesticides, food additives, and mycotoxins seriously threaten people's health and life safety. Chloramphenicol, clenbuterol hydrochloride, diethylstilbestrol, nitrofurans, and malachite green are key monitoring objects. In addition, illicit drugs, drugs of abuse such as morphine are extremely harmful. 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 hazardous substances are all small molecular compounds, and conventional detection methods are mainly divided into two categories: physical and chemical analysis methods and immunological anal...

Claims

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Application Information

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