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Method of 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 rapid on-site detection, short detection time and improved sensitivity Effect

Active Publication Date: 2019-11-19
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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  • Method of detecting Vibrio parahaemolyticus
  • Method of detecting Vibrio parahaemolyticus
  • Method of 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 metallomimetic 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 into 180mL deionized water, stir well until completely dissolved. Then heated to 50°C under nitrogen protection, the mixed solution was mixed with 12.5 mL 0.1M NH 3 ·H 2 O solution was mixed and stirred vigorously for 30 min. Under the action of an external magnetic field, wash with absolute ethanol and deionized water alternately until neutral, redissolve in aqueous solution, and store 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 solution was added dropwise...

Embodiment 2

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

[0034] (1) Preparation and modification of magnetic metallomimetic 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 into 180mL deionized water, stir well until completely dissolved. Then heated to 50°C under nitrogen protection, the mixed solution was mixed with 12.5 mL 0.1M NH 3 ·H 2 The O solution was mixed and stirred vigorously for 30 min. Under the action of an external magnetic field, wash with absolute ethanol and deionized water alternately until neutral, redissolve in aqueous solution, and store 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 solution was added dropw...

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Abstract

The invention relates to a method of detecting Vibrio parahaemolyticus, which comprises the following steps: (1) a test strip with a detection line coating a Vibrio parahaemolyticus antibody and a quality control line coating a goat anti mouse IgG antibody is prepared; (2) a magnet is placed below the detection area of the test strip; and (3) the content of the Vibrio parahaemolyticus is detected.Based on the intrinsic peroxidase mimicking enzyme properties of platinum-coated gold magnetic nanoparticles (Fe3O4@Au@Pt), the test strip can adopt a color method to determine the content of the Vibrio parahaemolyticus in a sample. The test strip has the advantages that the detection limit is low (10 cfu / mL), the specificity is good, the cost is low, the detection time is short and the like, canmeet the needs of rapid on-site detection, is suitable for detection on pathogenic bacteria and various toxic and harmful substances in liquid samples and is of great practical value.

Description

technical field [0001] The invention belongs to the field of analysis and detection, and in particular relates to a method for detecting vibrio parahaemolyticus. Background technique [0002] Vibrio parahaemolyticus, a Gram-negative bacillus, is one of the main pathogens threatening human health. Because it widely exists in seafood such as fish, shrimp, crab, shellfish and seaweed, the diseases caused by it often have a large number of people and are widely distributed, which poses 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, recombinant enzyme polymerase amplification method, real-time polymerase chain reaction method, enzyme-linked immunosorbent assay method, surface plasmon resonance method, quartz crystal microbalance method, etc. Sensors, Impedance Immunosensor...

Claims

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Application Information

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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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