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Vibrio parahaemolyticus detection method based on graphene oxide/ferroferric oxide/colloidal gold composite nanoparticle enhanced Raman effect

A technology of composite nanoparticles and hemolytic vibrio, applied in Raman scattering, measurement devices, instruments, etc., can solve the problems of detection time, sensitivity and specificity, cumbersome plate counting method, and low sensitivity. The effect of high surface-enhanced Raman activity, easy labeling, and high sensitivity

Inactive Publication Date: 2017-03-15
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The inspection procedure of the plate counting method is cumbersome, and there are defects in the detection time, sensitivity and specificity, etc.
Although the newly developed real-time fluorescent quantitative PCR technology can shorten the inspection time, it needs to extract the total bacterial DNA in the early stage, and the sensitivity is still not high; immunological methods such as enzyme-linked immunosorbent assay, electrochemical immunoassay, enzyme-linked fluorescence Analytical methods, etc., have the advantages of strong specificity, high sensitivity, and easy observation, but the cost of preparing antibodies is high, the cycle is long, and the stability is not good

Method used

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  • Vibrio parahaemolyticus detection method based on graphene oxide/ferroferric oxide/colloidal gold composite nanoparticle enhanced Raman effect
  • Vibrio parahaemolyticus detection method based on graphene oxide/ferroferric oxide/colloidal gold composite nanoparticle enhanced Raman effect
  • Vibrio parahaemolyticus detection method based on graphene oxide/ferroferric oxide/colloidal gold composite nanoparticle enhanced Raman effect

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Embodiment 1

[0023] 1) Synthesis of Raman-enhanced substrates

[0024] First prepare the aminated Fe 3 o 4 Nanoparticles: Weigh 0.9g of ferric chloride and dissolve it in 28mL of ethylene glycol, add 2.4g of sodium acetate and 0.696g of sodium dodecylsulfonate, stir at room temperature for 30min, then transfer to a reaction kettle at 200°C for 8h , after natural cooling, take it out, wash it with ethanol three times, and dry it in an oven at 50°C for 10 hours. Take 30mg ground Fe 3 o 4 Dissolve the particles in 20 mL of ethanol, then add 1 mL of 3-aminopropyltriethoxysilane dropwise, stir at room temperature for 6 hours, discard the supernatant by magnetic separation, add an appropriate amount of absolute ethanol, and ultrasonically disperse and wash. That is to prepare aminated Fe 3 o 4 nanoparticles.

[0025] Prepare nano-gold particles then: first get 49mL ultrapure water and add 0.5mL 1% chloroauric acid solution and stir and boil for 10min, then add 0.5mL 5% sodium citrate solu...

Embodiment 2

[0033] 1) Synthesis of Raman-enhanced substrates

[0034] First prepare the aminated Fe 3 o 4 Nanoparticles: Weigh 0.9g of ferric chloride and dissolve it in 28mL of ethylene glycol, add 2.4g of sodium acetate and 0.696g of sodium dodecylsulfonate, stir at room temperature for 30min, then transfer to a reaction kettle at 200°C for 8h , after natural cooling, take it out, wash it with ethanol three times, and dry it in an oven at 50°C for 10 hours. Take 30mg ground Fe 3 o 4 Dissolve the particles in 20 mL of ethanol, then add 1 mL of 3-aminopropyltriethoxysilane dropwise, stir at room temperature for 6 hours, discard the supernatant by magnetic separation, add an appropriate amount of absolute ethanol, and ultrasonically disperse and wash. That is to prepare aminated Fe 3 o 4 nanoparticles.

[0035] Prepare nano-gold particles then: first get 49mL ultrapure water and add 0.5mL 1% chloroauric acid solution and stir and boil for 10min, then add 0.5mL 5% sodium citrate solu...

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Abstract

The invention discloses a vibrio parahaemolyticus detection method based on a graphene oxide / ferroferric oxide / colloidal gold composite nanoparticle enhanced Raman effect. A graphene oxide / ferroferric oxide / colloidal gold (GO / Fe3O4@Au) composite nanoparticle is used as an active enhanced base, and a sulfhydrylization modified vibrio parahaemolyticus aptamer is added into the prepared GO / Fe3O4@Au composite nanoparticle, so that the vibrio parahaemolyticus aptamer is fixed on the base. A detected matter is mixed with the GO / Fe3O4@Au composite nanoparticle modified by the vibrio parahaemolyticus aptamer, and then the other vibrio parahaemolyticus aptamer modified by a Raman signal TAMRA is used for hatching; and Raman spectrum detection is carried out. By specifically combining the aptamer with vibrio parahaemolyticus, the detection of the vibrio parahaemolyticus in foods is realized. The method is high in sensitivity, high in specificity and convenient to operate, and has a wide application prospect in the field of food safety detection.

Description

technical field [0001] The invention relates to a detection method for vibrio parahaemolyticus based on graphene oxide / ferric oxide / colloidal gold composite nanoparticle enhanced Raman effect, which belongs to the field of food safety detection. Background technique [0002] Vibrio parahaemolyticus is a halophilic, Gram-negative rod-shaped bacterium that is widely distributed in the marine environment and in a variety of marine products. Eating raw or undercooked seafood infected with Vibrio parahaemolyticus can cause sudden food poisoning, abdominal pain, diarrhea, headache, nausea, vomiting and other symptoms, and severe patients may also experience dehydration, shock, even death. This pathogen is one of the main causes of foodborne illnesses in many Asian countries including China and Japan. According to data from my country's Foodborne Disease Surveillance Network, Vibrio parahaemolyticus food poisoning ranks first among microbial foodborne diseases in coastal and rive...

Claims

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Application Information

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IPC IPC(8): G01N21/65
CPCG01N21/658
Inventor 段诺吴世嘉沈默斐王周平
Owner JIANGNAN UNIV
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