Triple drug fluorescence immunochromatography reagent paper, detection kit and detection method

A drug and detection line technology, which is applied in the field of drug analysis and detection, can solve the problems of long time for displaying results, large differences between batches, and low analysis efficiency, and achieve the effect of saving the total time of detection, long detection time, and high analysis cost

Pending Publication Date: 2021-09-07
JINAN UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, ELISA detection still needs to be performed by professionals, and it takes a long time to display the results; colloidal gold immunochromatography technology can only perform qualitative analysis on samples with high content of drug metabolites such as urine, blood or saliva, which has low sensitivity and batch-to-batch differences problems such as large size and low analysis efficiency

Method used

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  • Triple drug fluorescence immunochromatography reagent paper, detection kit and detection method
  • Triple drug fluorescence immunochromatography reagent paper, detection kit and detection method
  • Triple drug fluorescence immunochromatography reagent paper, detection kit and detection method

Examples

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Embodiment 1

[0054] A magnetic fluorescent probe freeze-dried powder, prepared by the following steps:

[0055] (1) Activation: wash 100 μg of magnetic fluorescent microspheres with MES buffer (0.1M, pH 6.0) for 3 times, then resuspend in 990 μl of MES buffer solution, shake and mix well, and then add 10 μL of 10 mg / mL EDC respectively (carbodiimide) and 10 μL of 10 mg / mL NHS (N-hydroxysuccinimide), swirl at room temperature for 30 min; centrifuge at 15000 rpm for 30 min and discard the supernatant;

[0056] The magnetic fluorescent microspheres are prepared according to the literature (Guo L, Shao Y N, Duan H, et al.MagneticQuantum Dot Nanobead-Based Fluorescent Immunochromatographic Assay for the Highly Sensitive Detection of Aflatoxin B-1in Dark Soy Sauce[J].AnalyticalChemistry, 2019 , 91(7):4727-4734.) method, the fluorescence emission wavelength is 600-620nm, preferably 618nm.

[0057] (2) Antibody labeling: add 1000 μL of MES buffer to the pellet, resuspend by sonication; add 2 μg o...

Embodiment 2

[0067] A triple drug fluorescent immunoassay card, the structure of which is as follows: figure 1 As shown, including upper 10 and lower 9 jamming;

[0068] The upper cartridge 10 is provided with a sampling hole 12 and an observation hole 11;

[0069] The lower jam shell 9 is covered with a PVC base plate 1, and the base plate is a triple drug fluorescent immunochromatographic test strip;

[0070] The reagent paper is composed of a sample pad 2, an NC film 3 and an absorbent pad 4;

[0071] The NC membrane 3 is sequentially provided with a MET detection line 5, a MOR detection line 6, a KET detection line 7 and a quality control line 8 from an end close to the sample pad 2; the distance between the detection lines and the quality control lines is 3.5 mm, the distance between the KET detection line and the absorbent pad is 11mm;

[0072] The sample pad 2 of the reagent paper 3 overlaps with the sampling hole 12 of the upper cartridge 10 in position, which is convenient for ...

Embodiment 3

[0081] A triple drug fluorescent immunoassay kit, comprising the magnetic fluorescent probe freeze-dried powder described in embodiment 1 and the triple drug fluorescent immunoassay card of embodiment 3.

[0082] The method for carrying out fluorescent immunoassay detection of triple drugs in sewage based on the kit comprises the following steps:

[0083] (1) Mix 1 mL of sewage sample to be tested with 0.4 μg of magnetic fluorescent probe freeze-dried powder, incubate at 37°C for at least 10 minutes, and magnetically recover MFB-MET-mAbs, MFB-MOR-mAbs, and MFBs-KET in the sample -Immune complexes formed by mAbs and corresponding drug metabolites and unbound magnetic fluorescent probes; after removing the supernatant, resuspend the immune complexes in 100 μL PBS buffer to obtain the sample solution to be tested;

[0084](2) Take 75 μL of the sample solution to be tested and add it dropwise to the spotting hole of the triple drug fluorescence immunoassay card, and after reacting...

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Abstract

The invention discloses triple drug fluorescence immunochromatography reagent paper, a detection kit and a detection method. The triple drug fluorescence immunochromatography test strip consists of a sample pad, an NC (nitrocellulose) membrane and a water absorption pad, the NC membrane is sequentially provided with an MET detection line, an MOR detection line, a KET detection line and a quality control line from one end close to the sample pad. During detection, a to-be-detected sewage sample and the magnetic fluorescent probe freeze-dried powder are subjected to incubation reaction, recovered and added into a sample application hole of the detection card, and after chromatography, an observation hole of the detection card is scanned by a fluorescence reader; and the concentrations of the three drug metabolites are calculated according to the standard curve and the ratio of the electric signals of each detection line to the quality control line. When a sample is detected, a to-be-detected substance is captured and enriched by utilizing a magnetic response function of the magnetic fluorescent probe, so that interference of other substances is reduced; and a plurality of to-be-detected substances in the sample are quantitatively detected by using the multi-joint detection test strip. The sensitivity can reach ng/L, and the traceability requirements of drug metabolites in sewage at all levels are met.

Description

technical field [0001] The invention belongs to the field of drug analysis and detection, and in particular relates to a triple drug (methamphetamine MET, morphine MOR, ketamine KET) fluorescent immunochromatographic reagent paper, a detection kit and a detection method. Background technique [0002] At present, methamphetamine MET, morphine MOR, and ketamine KET are the three most widely abused drugs. [0003] The monitoring of illicit drug abuse is generally carried out through social epidemiological surveys, which have great limitations and uncertainties. [0004] Since the concept of sewage epidemiology was proposed in 2001, the technology of poison analysis based on sewage epidemiology has gradually been applied to the detection of drug abuse. The main sources of drugs and their metabolites in sewage are the excrement of drug addicts, the drugs washed away by drug addicts through the sewer to destroy evidence during raids, and the wastewater discharged from drug manufa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/558G01N33/58
CPCG01N33/54326G01N33/54346G01N33/558G01N33/588G01N33/5308
Inventor 唐勇徐涛滕佩君
Owner JINAN UNIVERSITY
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