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Preparation method and application of food-borne pathogen immunosensor based on gold label silver stain signal amplification technology

A food-borne pathogenic bacteria, immune sensor technology, applied in the direction of instruments, scientific instruments, measuring devices, etc., to achieve the effect of high sensitivity

Active Publication Date: 2016-08-24
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are no related reports on the preparation method and application of foodborne pathogenic bacteria immunosensor based on the combination of gold standard silver staining technology and multifunctional carbon nitride material

Method used

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  • Preparation method and application of food-borne pathogen immunosensor based on gold label silver stain signal amplification technology
  • Preparation method and application of food-borne pathogen immunosensor based on gold label silver stain signal amplification technology
  • Preparation method and application of food-borne pathogen immunosensor based on gold label silver stain signal amplification technology

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specific Embodiment 1

[0033] A method for preparing an immune sensor for food-borne pathogenic bacteria based on gold-labeled silver staining signal amplification technology, specifically comprising the following steps:

[0034] (1) Silanization of glass slides

[0035] Place the glass slide in piranha washing solution, heat it at 90-100 °C for 20-40 min, take it out, wash it three times with distilled water, and place it under N 2 After drying in the atmosphere, immerse in a silylating agent for silanization treatment at room temperature for 2-6 h, then wash with ethanol, and vacuum-dry at 110-130 °C for 1-3 h to obtain a silanized glass slide;

[0036] (2) Multifunctional carbon nitride material (Au@g-C 3 N 4 ) preparation method

[0037] a. Heat melamine at 500-600 °C for 3-5 h, and dry it in vacuum to obtain carbon nitride (g-C 3 N 4 ) powder; take 0.8-1.5 g of carbon nitride powder and add it to 80-120 mL of 4-6 mol / L HNO 3 solution, refluxed at 120-150 °C for 24-48 h, cooled naturally t...

specific Embodiment 2

[0045] The method for detecting food-borne pathogenic bacteria using the food-borne pathogenic bacteria immune sensor based on the gold-labeled silver staining signal amplification technology prepared in the above-mentioned specific example 1, the sample of the above-mentioned food-borne pathogenic bacteria immune sensor is placed on the Add 3-5 uL of silver staining reagent dropwise, react in the dark at room temperature for 10-15 min, wash off excess silver staining reagent with ultrapure water, and then dry in air, and dry the gold-labeled silver-stained sample on the glass slide Scan the point into a picture to obtain the gold-labeled silver-stained signal and its gray value average of the sample solution to be tested. According to the quantitative relationship between the gold-labeled silver-stained signal gray value and the concentration of the food-borne pathogenic bacteria solution, calculate the Measure the accurate concentration of foodborne pathogenic bacteria in the...

specific Embodiment 3

[0046] The preparation and detection application of the Vibrio parahaemolyticus immunosensor based on the gold-labeled silver staining signal amplification technology specifically includes the following steps:

[0047] (1) Silanization of glass slides

[0048] The slides were placed in piranha washing solution (obtained by mixing 98% concentrated sulfuric acid and 30% hydrogen peroxide at a volume ratio of 7:3), heated at 95 °C for 30 min, removed, washed three times with distilled water, and placed in N 2 After drying in the atmosphere, immerse in a silylating agent for silanization treatment at room temperature for 2-6 hours, then wash with ethanol, and vacuum-dry at 120 °C for 2 hours to obtain a silanized glass slide;

[0049] (2) Preparation method of multifunctional carbon nitride material

[0050] a. Heat melamine at 550 °C for 4 h and dry it in vacuum to obtain carbon nitride powder; add 1.2 g of carbon nitride powder to 100 mL 5 mol / L HNO 3 solution, refluxed at 135...

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Abstract

The invention discloses a preparation method and application of a food-borne pathogen immunosensor based on the gold label silver stain signal amplification technology. The preparation method is characterized by including the steps that a glass slide is silanized; a carbon nitride material is subjected to carboxylation and then mixed with a colloidal gold solution, the mixture is stirred, and a multifunctional carbon nitride solution is obtained; the multifunctional carbon nitride solution is combined with a food-borne pathogen primary antibody, and a food-borne pathogen primary antibody probe is obtained; a coupling reagent, a food-borne pathogen secondary antibody, a food-borne pathogen antigen and the food-borne pathogen primary antibody probe are sequentially dripped onto the silanized glass slide, and the food-borne pathogen immunosensor based on the gold label silver stain signal amplification technology is obtained. Quantitative detection and analysis are carried out by adding a silver stain reagent, and the preparation method and application of the food-borne pathogen immunosensor have the advantages that specificity and sensitivity are high, the result is accurate and reliable, cost is low, and operation is easy and fast.

Description

technical field [0001] The invention relates to a detection technology of food-borne pathogenic bacteria, in particular to a preparation method and application of an immune sensor for food-borne pathogenic bacteria based on gold-labeled silver staining signal amplification technology. Background technique [0002] Foodborne diseases are diseases caused by ingesting pathogenic agents in food. Foodborne diseases mainly include: food poisoning, intestinal diseases infected by food, foodborne parasitic diseases, and toxic diseases caused by toxic and harmful pollutants in food. The main pathogens in foodborne diseases are bacteria and their toxins. Foodborne pathogenic bacteria refer to pathogenic bacteria that cause bacterial food poisoning, such as Escherichia coli, Vibrio parahaemolyticus, Staphylococcus, Salmonella, Proteus Wait. Most of the diseases caused by these pathogenic bacteria occur in summer and autumn, which is consistent with the living habits of bacteria, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/532
CPCG01N33/532G01N33/56911G01N33/56916G01N33/56938
Inventor 宋信信郭智勇苏秀榕胡宇芳俞欣辰郭富米
Owner NINGBO UNIV
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