Listeria monocytogenes detection test strip based on hemolysin O as well as preparation and application of listeria monocytogenes detection test strip

A technology of Listeria monocytogenes and Listeria monocytogenes is applied to the detection test strip of Listeria monocytogenes based on hemolysin O and the field of preparation and application thereof, which can solve the problems of false negative detection cycle and false positive, and achieves Simple operation and low cost effect

Active Publication Date: 2021-03-30
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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  • Listeria monocytogenes detection test strip based on hemolysin O as well as preparation and application of listeria monocytogenes detection test strip
  • Listeria monocytogenes detection test strip based on hemolysin O as well as preparation and application of listeria monocytogenes detection test strip
  • Listeria monocytogenes detection test strip based on hemolysin O as well as preparation and application of listeria monocytogenes detection test strip

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 monocytogenes hemolysin O (LLO) as well as a preparation method and application of the Listeria monocytogenes detection test strip. The detection method relates to preparation calibration or calibration of the Listeria monocytogenes detection test strip based on LLO. The concentration of the Listeria monocytogenes in the milk or instant meat sample is detected by using the calibrated Listeria monocytogenes detection test strip through a labeling recovery method. The detection test strip and the method for detecting Listeria monocytogenes by using the detection test strip provided by the invention have the advantages of intuition, accuracy, simplicity and convenience in operation, short detection time, low cost, wide application range, simplicity in storage, long shelf life, easiness in popularization and use and the like, and have a wide application prospect.

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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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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