Phenobarbital homogeneous-phase enzyme immunoassay reagent kit and preparation method thereof

A detection kit and homogeneous enzyme immunization technology, applied in the field of medical testing, can solve the problems of only semi-quantitative, complicated operation and high cost, and achieve the effects of high precision, strong specificity and high accuracy

Active Publication Date: 2012-01-18
西安金域医学检验所有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented method allows for accurate measurement of certain substances called phobionuclidine or its analogues such as benthazoxane by measuring their concentrations accurately with an assortment of different reagents. It can be used at home without complicated equipment like electronic counters but it provides more precise results compared to existing methods due to better control over reaction conditions during analysis.

Problems solved by technology

This patented technical solution involves analyzing different types of substances called phenobarbazone or its analogues that are found naturally occurring throughout human body by various techniques such as High Performance Liquor Chromatochromatic Immune Assays(HPICA). These tests provide accurate data on how well certain medications work effectively when given through their regular dosage form. However, these current analysis tools have limitations: they require complex operations, take longer periods, involve higher costs than traditional treatments, cannot detect non-therapy agents like antiarrhythmics without being able to determine the exact amount needed based solely upon previous results obtained from earlier trials conducted under similar conditions.

Method used

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  • Phenobarbital homogeneous-phase enzyme immunoassay reagent kit and preparation method thereof
  • Phenobarbital homogeneous-phase enzyme immunoassay reagent kit and preparation method thereof
  • Phenobarbital homogeneous-phase enzyme immunoassay reagent kit and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0037] a. Synthesis of bovine serum albumin-phenobarbital derivative immune antigen:

[0038] 1. Preparation of BSA solution, weigh 20 mg of bovine serum albumin (BSA) and dissolve it in 5 mL of 0.2 mol / L, pH 8.5 phosphate buffer at room temperature;

[0039] 2. Activation of phenobarbital derivatives, weigh 10 mg of specific phenobarbital derivatives in a small beaker, and add 350 μL dimethylamide (DMF), 350 μL ethanol, 700 μL 10 mmol / L, pH5.0 Potassium phosphate buffer, 40 mg 1-ethyl-3-(-3-dimethylaminopropyl) carbodiimide, and 5 mg N-hydroxysulfosuccinimide (NHS), these chemicals were stirred and dissolved at room temperature React for 30 minutes;

[0040] 3. Coupling and purification of the antigen. The activated phenobarbital derivative is added dropwise to the BSA solution and stirred overnight at 2-8° C. to obtain the antigen; and the coupled antigen is dialyzed and purified.

[0041] B, the preparation of anti-phenobarbital derivative antibody, the steps are as follo...

Embodiment 2

[0052] The detection experiment of embodiment diphenobarbital detection kit product

[0053] 1. Calibration experiment

[0054] see figure 2 , the phenobarbital calibrator was dissolved in blank serum and prepared into 6 kinds of calibrating solutions with a series concentration of 0, 5, 10, 20, 40, and 80 μg / mL. The working volume of the calibrating solution was 10-35 μL, and then Add 100-200 μL of reagent R1 and 100-200 μL of reagent R2, and use the two-point rate method to detect the absorbance change rate at the main wavelength of 340 nm and the secondary wavelength of 405 nm, and establish and optimize the phenobarbital homogeneous enzyme immunoassay calibration curve ( figure 2 ), the establishment and optimization of the calibration curve was completed on a Hitachi 7180 automatic biochemical analyzer.

[0055] 2. Recovery experiment

[0056] Using the established phenobarbital calibration curve, determine low (20 μg / mL), medium (40 μg / mL)) and high (80 μg / mL) serum...

Embodiment 3

[0077] Embodiment 3 Utilize automatic biochemical analyzer to carry out sample test

[0078] (1). Collection of serum samples: collect serum samples according to conventional methods;

[0079] (2). According to the operating instructions of the Hitachi 7180 automatic biochemical analyzer, turn on the instrument, perform the optical density detection of the instrument and clean the probe, and check whether the instrument is operating normally;

[0080] (3). After the detection of the instrument is running normally, put the reagents R1 and R2 into the reagent chambers of R1 and R2 in turn, put the serum sample into the sample tray 1 (S1), put 0μg / mL, 5μg / mL, 10μg / mL, 20μg / mL Put the phenobarbital calibration solution of mL, 40 μg / mL, and 80 μg / mL into the designated position of the sample tray 2 (S2);

[0081] (4). When the instrument is in the Stand by state, set the operating procedures and detection parameters of phenobarbital, and the specific detection parameters are shown...

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Abstract

The invention aims at providing a phenobarbital drug concentration testing device with the advantages of simpleness, convenience, quickness, high sensitivity and full automation and a preparation method thereof. In order to solve the abovementioned technical problems, the invention provides a phenobarbital homogeneous-phase enzyme immunoassay reagent kit and preparation method thereof, and the preparation method comprises synthesis of phenobarbital immunogens, preparation of anti-phenobarbital specific antibodies, preparation of enzyme-labeled conjugates and determination of a sample. The phenobarbital drug concentration homogeneous-phase enzyme immunoassay reagent kit provided by the invention has high accuracy and precision, the average recovery rate of serum specimens is larger than 95percent, and the related coefficient deviation is less than 2.5 percent. The specificity of phenobarbital antibodies provided in the reagent kit is strong; and during a test of drug-to-drug interactions, nearly no interaction among thirty tested common drugs and compounds occurs, and the phenobarbital drug concentration homogeneous-phase enzyme immunoassay reagent kit is suitable for clinically testing the concentration of the phenobarbital drug in the blood and has the advantages of simpleness, convenience, quickness, low cost, high sensitivity and full automation.

Description

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Claims

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Application Information

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Owner 西安金域医学检验所有限公司
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