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A test strip for detecting Listeria monocytogenes based on hemolysin O and its preparation and application

A Listeria monocytogenes, Listeria monocytogenes technology, applied in the hemolysin O-based Listeria monocytogenes detection test strip and its preparation and application field, can solve the problems of false positive and false negative detection cycle, etc., to achieve The effect of long shelf life, simple storage, and easy large-scale promotion and use

Active Publication Date: 2021-11-05
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods also have problems such as false positive or false negative probability and long detection cycle.

Method used

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  • A test strip for detecting Listeria monocytogenes based on hemolysin O and its preparation and application
  • A test strip for detecting Listeria monocytogenes based on hemolysin O and its preparation and application
  • A test strip for detecting Listeria monocytogenes based on hemolysin O and its preparation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1: Preparation of a test strip for detecting Listeria monocytogenes based on LLO

[0044] (1) Preparation of nanoporous gold (NPG): Au / Ag (Au 48 Ag 52 , wt%) alloy sheet is placed in concentrated nitric acid, at 40 ± 2 ℃ temperature, corrosion 2 hours, makes NPG thin film;

[0045] (2) Prepare NPG / CWE electrode: prepare a test strip with 1 working electrode (carbon working electrode, CWE), an Ag / AgCl reference electrode and a carbon counter electrode, and fix the prepared NPG film on the test strip On the working electrode CWE, prepare NPG / CWE electrode;

[0046] (3) Preparation of CAT-Lipo / NPG / CWE bioelectrode: phosphatidylcholine, cholesterol and 1-octadecanethiol (molar ratio 10:10:1) were mixed in chloroform and placed in a small mouth bottle. then use N 2 The organic solvent was removed to form a thin lipid layer at the bottom. Add an appropriate amount of 0.1M catechol solution to make a final lipid concentration of 0.5mg mL -1 , and then ultrason...

Embodiment 2

[0047] Embodiment 2: A kind of LLO-based Listeria monocytogenes detection test strip preparation

[0048] (1) Preparation of nanoporous gold (NPG): Au / Ag (Au 50 Ag 50 , wt%) alloy sheet is placed in concentrated nitric acid, at 35 ± 2 ℃ temperature, corrodes 0.5 hour, makes NPG thin film;

[0049] (2) Prepare NPG / CWE electrode: prepare a test strip with 1 working electrode (carbon working electrode, CWE), an Ag / AgCl reference electrode and a carbon counter electrode, and fix the prepared NPG film on the test strip On the working electrode CWE, prepare NPG / CWE electrode;

[0050] (3) Preparation of CAT-Lipo / NPG / CWE bioelectrode: Phosphatidylcholine, cholesterol and 1-octadecanethiol were mixed in chloroform at a ratio of 9:9:2 and placed in a vial. then use N 2 The organic solvent was removed to form a thin lipid layer at the bottom. Add an appropriate amount of 0.6M catechol solution to make a final lipid concentration of 4.0mg mL -1 , and then ultrasonically for 20 minu...

Embodiment 3

[0051] Embodiment 3: A kind of LLO-based Listeria monocytogenes detection test strip preparation

[0052] (1) Preparation of nanoporous gold (NPG): Au / Ag (Au 40 Ag 60 , wt%) alloy sheet is placed in concentrated nitric acid, at 45 ± 2 ℃ temperature, corrodes 4 hours, makes NPG thin film;

[0053](2) Prepare NPG / CWE electrode: prepare a test strip with 1 working electrode (carbon working electrode, CWE), an Ag / AgCl reference electrode and a carbon counter electrode, and fix the prepared NPG film on the test strip On the working electrode CWE, prepare NPG / CWE electrode;

[0054] (3) Preparation of CAT-Lipo / NPG / CWE bioelectrode: phosphatidylcholine, cholesterol and 1-octadecanethiol were mixed in chloroform according to the molar ratio of 9-10:9-10:1-2, Place in a vial. then use N 2 The organic solvent was removed to form a thin lipid layer at the bottom. Add an appropriate amount of 1.5M catechol solution to make a final lipid concentration of 8.0mg mL -1 , and then ultra...

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Abstract

The invention discloses a Listeria monocytogenes detection test strip based on Listeria hemolysin O (LLO) and its preparation method and application. The detection method relates to the preparation of the LLO-based Listeria monocytogenes detection test strip Calibration or calibration; use the calibrated Listeria monocytogenes detection test strip to detect the concentration of Listeria monocytogenes in milk or ready-to-eat meat samples by using the standard recovery method. The detection test strip and the method for detecting Listeria monocytogenes provided by the present invention have the advantages of intuition and accuracy, simple and convenient operation, short detection time, low cost, wide application range, simple storage, long shelf life, and easy popularization and use. application prospects.

Description

technical field [0001] The invention relates to a test strip for detecting Listeria monocytogenes and its preparation and application, in particular to a test strip for detecting Listeria monocytogenes based on Listeriolysin O (LLO) and its preparation and application. Preparation and application. Background technique [0002] Listeria monocytogenes (Listeria monocytogenes, referred to as Listeria monocytogenes) is a zoonotic pathogenic bacteria, which is highly resistant to low temperature, high salt and disinfectants. The fungus is a saprophyte that feeds on dead and decaying organic matter and is also a contaminant in certain foods (mainly fresh milk products) and can cause severe food poisoning. Listeriosis can be caused by ingestion of food contaminated with Listeria monocytogenes in humans. Listeriosis poses a greater risk to immunocompromised patients, pregnant women, and newborns. Because Listeria monocytogenes can cross the intestinal barrier, blood-brain barrier...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/48G01N27/416G01N27/327G01N27/30
CPCG01N27/301G01N27/308G01N27/3277G01N27/3278G01N27/4163G01N27/48
Inventor 王霞张亚超许平
Owner SHANDONG UNIV
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