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Methadone immunodetection reagent and preparation and detection methods

A technology of immune detection and detection method, which is applied in the field of medical inspection, can solve the problems of low measurement accuracy and complicated operation of methadone detection process, and achieve the effects of high sensitivity, accurate quantification and strong specificity

Inactive Publication Date: 2018-03-23
太原瑞盛生物科技有限公司
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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 methadone detection process in the prior art. The present invention provides a fast, high-sensitivity, and accurate detection of the methadone content in the sample to be tested. Homogeneous enzyme immunoassay Detection reagent and preparation method thereof

Method used

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  • Methadone immunodetection reagent and preparation and detection methods
  • Methadone immunodetection reagent and preparation and detection methods

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Preparation of glucose dehydrogenase-antigen conjugate

[0046] 1) Preparation of methadone hapten

[0047] a. Dissolve 3 g of methadone in 35 mL of anhydrous pyridine, add 35 mL of acetic anhydride, stand at room temperature for 16 hours, concentrate under reduced pressure, evaporate to dryness and weigh to obtain product A;

[0048] b. Dissolve 40 g of product A in 50 mL of dimethylformamide, then add 15 mL of 1M hydrochloric acid, let stand at room temperature for 16 hours, then extract with 50 mL of ether, collect the organic phase and extract with 50 mL of sodium hydroxide aqueous solution , continue to collect the organic phase and concentrate under reduced pressure, add 15 mL of petroleum ether, and evaporate to dryness to obtain product B;

[0049] c. Dissolve an appropriate amount of product B in 30 mL of acetic anhydride, react at 94°C for 20 minutes, cool and crystallize overnight, and evaporate to dryness under reduced pressure to obtain product ...

Embodiment 2

[0058] Example 2: Preparation of Methadone Homogeneous Enzyme Immunoassay Reagent

[0059] Homogeneous enzyme immunoassay reagents for methadone, including: enzyme-labeled methadone, indicator reagent for detecting methadone antibody-enzyme-labeled methadone complex. The indicator reagent is selected from enzyme reagents, radioisotope reagents, fluorescent reagents and chemiluminescent 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.

[0060] Before using the methadone homogeneous enzyme immunoassay reagent, in order to avoid the reaction between the enzyme-labeled conjugate and the enzyme substrate in the indicator reagent, the enzyme-labeled conjugate and the enzyme substrate are placed separately, so the methadone homogeneous enzyme The immunoassay reagents cons...

Embodiment 3

[0069] Embodiment three: Methadone homogeneous enzyme immunoassay reagent reaction calibration curve

[0070] 1) Calibrator preparation: Dissolve commercially available human methadone 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. Taking the methadone calibrator of Beijing Cool Technology Co., Ltd. as the original standard, the methadone kit was used to test the calibrator of different concentrations 10 times, and the average value was obtained to obtain the concentration of the methadone calibrator: 10, 25, 50, 100, 400 , 800 ng / mL.

[0071] 2) Biochemical analyzer detection: Take the operation of Hitachi 7170 as an example: the measurement wavelength is 340 nm, take different concentrations of calibrator solutions (15 μL), add methadone R 1 Reagent (200 μL), mix well, then add methadone R 2 Reagent (50 μL), after mixing, measure the OD at different time...

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Abstract

The invention discloses a methadone immunodetection reagent and a preparation and detection method thereof. Including: enzyme-labeled methadone, an indicator reagent for detecting methadone antibody-enzyme-labeled methadone complex; the above-mentioned enzyme-labeled methadone is formed by coupling methadone and glucose dehydrogenase. The methadone immunoassay reagent of the invention can accurately and rapidly determine the content of methadone in samples such as human blood. Compared with the existing detection reagents on the market, the detection reagent of the present invention has the advantages of convenience and rapidity, high sensitivity, strong specificity, accurate quantification, etc., and is beneficial to clinical popularization and use.

Description

technical field [0001] The invention relates to the field of medical testing, in particular to a methadone immunodetection reagent and a preparation and detection method thereof. Background technique [0002] Methadone (MTD) is a synthetic opioid drug that is similar in efficacy to morphine, has analgesic effects, and is accompanied by side effects such as respiratory depression, miosis, and sedation. Methadone has strong analgesic effect, long duration, and good oral effect. It is suitable for chronic pain and is the main drug in the substitution therapy for opioid dependence in the world. However, long-term repeated use can produce long-term clinical effects, which can form physical dependence, Psychological dependence and tolerance. Therefore, in the application of methadone treatment, the addiction and abuse tendency of methadone should not be ignored, and a strict management system must be established to avoid loss and transformation into drugs. [0003] Currently, th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573
CPCG01N33/573
Inventor 康志禹丁兰艳赵慧敏严芳芳杜爱铭徐兵
Owner 太原瑞盛生物科技有限公司
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