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A method for detecting vibrio parahaemolyticus

A hemolytic vibrio and detection line technology, which is applied in the field of analysis and detection, can solve the problems of hindering the application of immunochromatography technology and the low sensitivity of mobile rate immunochromatography technology, and achieve the goals of short detection time, low detection limit and improved sensitivity Effect

Active Publication Date: 2022-05-27
QILU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In order to solve this bottleneck that hinders the application of immunochromatography technology, in the present invention, we replace the classically used colloidal gold with magnetic Fe 3 o 4 @Au@Pt nanoparticles, using an external magnetic field to reduce their movement rate on the nitrocellulose membrane overcomes the defect of low sensitivity of immunochromatography

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  • A method for detecting vibrio parahaemolyticus
  • A method for detecting vibrio parahaemolyticus
  • A method for detecting vibrio parahaemolyticus

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Embodiment 1

[0020] A method for detecting Vibrio parahaemolyticus, comprising the following steps:

[0021] (1) Preparation and modification of magnetic metal mimetic enzymes:

[0022] Fe 3 O 4 Preparation of nanoparticles: Accurately weigh 11.2 mM ferric chloride solution and 5.6 mM ferrous chloride solution, add them to 180 mL of deionized water, and stir well until completely dissolved. Then heated to 50 °C under nitrogen protection, the mixed solution was mixed with 12.5 mL of 0.1 M NH 3 ·H 2 The O solution was mixed and stirred vigorously for 30 min. Under the action of an external magnetic field, alternately rinsed with absolute ethanol and deionized water until neutral, redissolved in an aqueous solution, and stored at 4°C for later use;

[0023] Fe 3 O 4 Preparation of @Au nanoparticles: under ultrasonic conditions, 20 mL ① synthesized Fe 3 O 4 The solution was mixed with 15 mL of 1% chloroauric acid solution for 20 min. Next, 15 mL of 20 mM trisodium citrate soluti...

Embodiment 2

[0033] A method for detecting Vibrio parahaemolyticus, comprising the following steps:

[0034] (1) Preparation and modification of magnetic metal mimetic enzymes:

[0035] Fe 3 O 4 Preparation of nanoparticles: Accurately weigh 11.2 mM ferric chloride solution and 5.6 mM ferrous chloride solution, add them to 180 mL of deionized water, and stir well until completely dissolved. Then heated to 50 °C under nitrogen protection, the mixed solution was mixed with 12.5 mL of 0.1 M NH 3 ·H 2 The O solution was mixed and stirred vigorously for 30 min. Under the action of an external magnetic field, alternately rinsed with absolute ethanol and deionized water until neutral, redissolved in an aqueous solution, and stored at 4°C for later use;

[0036] Fe 3 O 4 Preparation of @Au nanoparticles: under ultrasonic conditions, 20 mL ① synthesized Fe 3 O 4 The solution was mixed with 15 mL of 1% chloroauric acid solution for 20 min. Next, 15 mL of 20 mM trisodium citrate solutio...

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Abstract

The invention relates to a method for detecting Vibrio parahaemolyticus, comprising the following steps: (1) preparing test strips in which the detection line is coated with Vibrio parahaemolyticus antibody and the quality control line is coated with goat anti-mouse IgG antibody; (2) ) Place a magnet under the detection area of ​​the test strip; (3) Detect the content of Vibrio parahaemolyticus. Based on platinum-coated gold magnetic nanoparticles (Fe 3 o 4 @Au@Pt) has intrinsic peroxidase-mimicking enzyme properties, and the test strip can use the color method to determine the content of Vibrio parahaemolyticus in the sample. The test strip of the present invention has the advantages of low detection limit (10cfu / mL), good specificity, low cost, short detection time, etc., can meet the needs of on-site rapid detection, and is suitable for pathogenic bacteria and various toxic and harmful substances in liquid samples The detection of substances is of great practical value.

Description

technical field [0001] The invention belongs to the field of analysis and detection, in particular to a method for detecting Vibrio parahaemolyticus. Background technique [0002] Vibrio parahaemolyticus, a gram-negative bacillus, is one of the major pathogens threatening human health. Due to its widespread presence in seafood such as fish, shrimp, crab, shellfish and seaweed, the diseases caused by it are often numerous and widely distributed, posing a huge threat to food safety and people's health. Therefore, the detection of Vibrio parahaemolyticus in food is of great significance. [0003] At present, the commonly used methods for detecting food-borne pathogens mainly include plate counting method, recombinase polymerase amplification method, real-time polymerase chain reaction method, enzyme-linked immunosorbent assay, surface plasmon resonance method, quartz crystal microbalance Sensors, impedance-type immunosensors, color sensors, fluorescence immunoassays, etc. Th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/558
CPCG01N33/56916G01N33/558
Inventor 吴正宗刘鹏飞崔波袁超于滨郭丽赵海波陶海腾
Owner QILU UNIV OF TECH
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