Amphetamine immunodetection reagent and preparation and detection methods thereof

An amphetamine and immunological detection technology, applied in the field of medical testing, can solve the problems of low measurement accuracy and complicated operation of amphetamine detection process, and achieve the effects of high sensitivity, accurate quantification and strong specificity

Inactive Publication Date: 2018-05-22
太原瑞盛生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the problems of complex operation and low measurement accuracy in the detection process of amphetamine in the prior art. Amphetamine homogeneous enzyme immunoassay reagent and preparation method thereof

Method used

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  • Amphetamine immunodetection reagent and preparation and detection methods thereof
  • Amphetamine immunodetection reagent and preparation and detection methods thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Preparation of glucose dehydrogenase-antigen conjugate

[0039] 1) Coupling of glucose dehydrogenase (GDH) with amphetamine

[0040] a. Dissolve 15 mg of amphetamine in 250 μL of dimethylformamide (DMF), shake slightly to dissolve; at the same time, dissolve 30.0 mg of GDH with a specification of 100 KU in 3 mL of 0.01mol / L PBS (pH7.4) In the buffer solution, shake and dissolve evenly;

[0041] b. Slowly add the amphetamine solution into the GDH solution while stirring, and slowly add 20 μL of 25% glutaraldehyde solution dropwise at the same time. After the dropwise addition, place the solution at 4°C and stir overnight.

[0042] 2) Purify the conjugated enzyme-labeled antigen

[0043] The conjugated enzyme-labeled antigen was purified by G-25 gel chromatography column to obtain glucose dehydrogenase-amphetamine conjugate, which was stored at 2-8°C.

Embodiment 2

[0044] Example 2: Preparation of Amphetamine Homogeneous Enzyme Immunoassay Reagent

[0045] Amphetamine homogeneous enzyme immunoassay reagents, including: enzyme-labeled amphetamine, indicator reagent for detecting amphetamine antibody-enzyme-labeled amphetamine complex. The indicator reagent is selected from enzyme reagents, radioisotope reagents, fluorescent reagents and chemiluminescence reagents. Preferably, the indicator reagent is an enzyme reagent, including: an enzyme-labeled conjugate and an enzyme substrate. Wherein, the enzyme-labeled conjugate includes glucose dehydrogenase-antigen conjugate, which can be obtained by chemical synthesis.

[0046] Before using the amphetamine homogeneous enzyme immunoassay reagent, in order to avoid the reaction between the enzyme-labeled conjugate in the indicator reagent and the enzyme substrate, the enzyme-labeled conjugate and the enzyme substrate are placed separately, so the safety Amphetamine homogeneous enzyme immunoassay...

Embodiment 3

[0055] Example 3: Amphetamine Homogeneous Enzyme Immunoassay Reagent Reaction Calibration Curve.

[0056] 1) Preparation of AMPH calibrator: Dissolve commercially available human amphetamine recombinant protein in a solution similar to human serum matrix (NaCl 0.9%, BSA 0.2%, NaN 3 0.1%, Tris-HCl pH 7.4), and made different concentrations of calibrators. Using the amphetamine calibration product of Taipu Biosciences (China) Co., Ltd. as the original standard, the amphetamine kit was used to test the calibration products of different concentrations for 10 times, and the average value was obtained to obtain the amphetamine calibration product Concentration: 10, 50, 100, 200, 400, 800ng / mL.

[0057] 2) Detection of biochemical analyzer: take the operation of Hitachi 7170 as an example: the measurement wavelength is 340nm, take different concentrations of calibrator solutions (15μl), add amphetamine R 1 Reagent (200 μl), mix well, then add amphetamine R 2 Reagent (50μl), after...

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Abstract

The invention discloses an amphetamine immunodetection reagent and preparation and detection methods thereof. The amphetamine immunodetection reagent comprises enzyme labeled amphetamine and an indication reagent for detecting an amphetamine antibody-enzyme labeled amphetamine compound, wherein the enzyme labeled amphetamine is formed by coupling amphetamine with glucose dehydrogenase. The amphetamine immunodetection reagent can accurately and quickly determine the content of amphetamine in such samples as human blood. Compared with a conventional detection reagent on the market, the amphetamine immunodetection reagent has the advantages of being convenient and quick, high in sensitiveness, strong in specificity and accurate in quantification, and facilitates clinical popularization.

Description

technical field [0001] The invention relates to the field of medical testing, in particular to an amphetamine immunodetection reagent and a preparation and detection method thereof. Background technique [0002] Amphetamine (AMPH), also known as amphetamine, is a central nervous system excitatory and antidepressant drug commonly used clinically. AMPH induces profound psychoactive effects, including increased alertness, initiative and confidence, decreased euphoria, decreased fatigue, increased speech, and increased ability to concentrate. AMPH can also cause adverse reactions, which can produce long-term damage to brain tissue, the most common being hyperexcitability, with symptoms such as restlessness, insomnia, tremor, nervousness and irritability. The most serious consequence of taking large doses of AMPH is a toxic neuropathy, so it is very important to monitor the patient's blood drug levels. [0003] At present, the commonly used methods for detecting amphetamine are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53
CPCG01N33/53
Inventor 马运乐杨钰祥于雪纯康志禹杜爱铭徐兵
Owner 太原瑞盛生物科技有限公司
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