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Imidaclothiz bioluminescence lateral flow immunochromatography

A technology of bioluminescence and immunochromatography, applied in the field of biology, can solve problems that have not yet been seen, and achieve the effects of reducing detection costs, high novelty, and high sensitivity

Inactive Publication Date: 2018-09-21
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the bioluminescent lateral flow immunochromatographic method based on mimetic epitope polypeptides and catalyzed luminescent enzyme recombinant expression tracers at home and abroad

Method used

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  • Imidaclothiz bioluminescence lateral flow immunochromatography
  • Imidaclothiz bioluminescence lateral flow immunochromatography
  • Imidaclothiz bioluminescence lateral flow immunochromatography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Detection of chlorothialine pesticide standard by luminescent lateral flow immunochromatography detection method

[0029] 1. Preparation of chlorothialine pesticide standard solution

[0030] Prepare chlorothialine standard stock solution (1 mg / mL) with methanol, and dilute the stock solution with PBS containing 5% methanol to a series concentration from 750 ng / mL to 0.01 ng / mL for detection by lateral flow immunochromatography.

[0031] 2. The recombinant tracer competes with the analyte for the binding site of the capture antibody

[0032] 50 μL of standard solution was mixed with 50 μL of recombinant tracer (containing 5 μg protein, 0.2% Tween 20 and 2 mg / mL BSA), and the mixture was added to the loading well and flowed through the membrane for 15 minutes.

[0033]3. Detection of Bioluminescent Signals

[0034] Add 100 μL PBS to wash the membrane for 10 minutes. Add 25 µL of assay buffer containing 2 µL of nano-luciferase assay substrate directly onto t...

Embodiment 2

[0042] Example 2: Luminescent lateral flow immunochromatographic detection method for detection of added samples

[0043] 1. Preparation and processing of added samples

[0044] The chlorothialine standard was added to the soil, orange and brown rice samples for addition recovery test. Weigh 10 g of crushed and mixed soil, orange and brown rice samples, add standard products to the final concentration of 20, 100 and 500 ng / g, mix well, let stand overnight at room temperature in the dark, add 20 mL of PBS buffer containing 25% methanol Mix well, vortex for 5 minutes, sonicate for 15 minutes, centrifuge at 4000rpm for 5 minutes, transfer all the supernatant to a 25mL volumetric flask, and dilute to 25mL with PBS buffer. Then dilute 4 times with PBS buffer, and then dilute 4 times with PBS buffer containing 5% methanol for detection.

[0045] 2. The recombinant tracer competes with the analyte for the binding site of the capture antibody

[0046] 50 μL of sample solution was m...

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Abstract

The invention relates to an imidaclothiz bioluminescence lateral flow immunochromatography. The imidaclothiz bioluminescence lateral flow immunochromatography includes the steps that analyte and recombination tracer jointly compete binding sites of antibody capturing on a nitrocellulose membrane; then the membrane is washed, and a reaction substrate is added to generate fluorescence signals; by combining a mobile-phone detection part, a smart phone is used for photographing a detection result, and finally, an image photographed by the mobile pone is analyzed and subjected to result determining. According to the imidaclothiz bioluminescence lateral flow immunochromatography, recombinant protein of phage-display-polypeptide simulation epitope and recombinant protein of catalytic luminescentenzyme are used as the tracer, and by combining the smart phone, detection of a small molecule compound is achieved through lateral flow chromatography mode. The imidaclothiz bioluminescence lateral flow immunochromatography is high in sensitivity, rapid, easy and convenient, economical and high in practicability, and field quantitative detection can be achieved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a chlorothialine bioluminescence lateral flow immunochromatography method, which uses phage to display polypeptide mimetic epitopes and recombinant proteins that can catalyze luminescent enzymes as tracers, cooperates with smart phones, and uses lateral flow Chromatography realizes the online and quantitative detection of small molecule compounds. Background technique [0002] Chlorthialine is a neonicotinoid insecticide independently developed by my country, which is widely used to control pests on crops such as wheat, rice, fruit trees and vegetables. However, excessive use makes it widely distributed in water bodies and agricultural products, posing a potential threat to non-target organisms such as bees. Therefore, establishing a simple, rapid and sensitive detection method to monitor chlorothialine residues in the environment and agricultural products is an important...

Claims

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Application Information

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IPC IPC(8): G01N33/558
CPCG01N33/558G01N2430/10
Inventor 王鸣华华修德丁园
Owner NANJING AGRICULTURAL UNIVERSITY
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