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Seafood product detection kit, preparation method and applications thereof

A detection kit and kit technology, which is applied in the field of microbial detection, can solve the problems of application limitations and low detection sensitivity of colloidal gold test strips, and achieve the effects of short detection time, increasing the amount of antibody labeling, and improving sensitivity

Inactive Publication Date: 2016-12-07
NINGBO INST OF MATERIALS TECH & ENG CHINESE ACADEMY OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the detection sensitivity of colloidal gold test strips is relatively low (usually higher than 10 4 CFU / mL), the application of this method is limited when detecting substances with low sensitivity requirements (Vibrio parahaemolyticus must not be detected in EU regulations)

Method used

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  • Seafood product detection kit, preparation method and applications thereof
  • Seafood product detection kit, preparation method and applications thereof
  • Seafood product detection kit, preparation method and applications thereof

Examples

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preparation example Construction

[0073] (3) Preparation of immunochromatographic test strips: Treat the sample pad with a certain treatment solution, such as pH 8.5 0.1M PBS buffer solution (0.5% BSA, 0.5% Tween-20), and place it in a 60°C blast drying oven After 3 hours, take it out and put it in a drying tank for later use; use the rabbit polyclonal antibody and donkey anti-mouse secondary antibody immunized with Vibrio parahaemolyticus at a certain concentration and spray volume, for example, the concentration is 1.0-1.5mg / mL, and the spray volume is uniform. Spray 0.75uL / cm on the detection pad (usually nitrocellulose membrane) as the detection line (T line) and quality control line (C line), respectively, and vacuum dry overnight at 30°C. Assemble the sample pad, bonding pad, NC membrane, and absorbent paper in a certain order, cut into 4mm wide strips, put them in a tin foil bag, add a desiccant and seal it, and store it in a desiccator for later use.

[0074] (4) Detection of samples by test strips: Ta...

Embodiment 1

[0077] Example 1: Detection of Vibrio parahaemolyticus in shrimp meat by nano-silver-gold core-shell particle immunochromatography system

[0078] (1) Preparation of nano-silver-gold core-shell particles: This experiment mainly has 2 steps. First, the synthesis of silver nanoparticles: get 100mL 5mM trisodium citrate solution and 0.1mM tannic acid solution at a constant temperature of 240-250°C Heat at reflux, add 1mL 25mM AgNO after boiling 3 Aqueous solution, when the color of the solution is stable, continue to stir for 3 minutes, stop heating, and cool to room temperature in the dark. Second, the synthesis of nano-silver-gold core-shell particles: take 100mL 10mg of HAuCl 4 The aqueous solution was heated under reflux at a constant temperature of 120-130 ° C. After 10 min, 4 mL of prepared silver nanoparticles and 400 uL of 1% (w / v) sodium citrate solution were added. After 30 min of reaction, the heating was stopped and cooled to room temperature. image 3 shown.

[00...

Embodiment 2

[0084] Example 2: Detection of Vibrio parahaemolyticus in fish by nano-silver-gold core-shell particle immunochromatography system

[0085] (1) Preparation of nano-silver-gold core-shell particles: This experiment mainly has 2 steps. First, the synthesis of silver nanoparticles: get 100mL 5mM trisodium citrate solution and 0.1mM tannic acid solution at a constant temperature of 240-250°C Heat at reflux, add 1mL 25mM AgNO after boiling 3 Aqueous solution, when the color of the solution is stable, continue to stir for 3 minutes, stop heating, and cool to room temperature in the dark. Second, second, the synthesis of nano-silver-gold core-shell particles: take 100mL 10mg of HAuCl 4 The aqueous solution was heated under reflux at a constant temperature of 120-130°C. After 10min, 4mL of prepared silver nanoparticles and 400uL of 1% (w / v) sodium citrate solution were added. After 30min of reaction, the heating was stopped and cooled to room temperature.

[0086] (2) Labeling of na...

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Abstract

The present invention discloses a seafood product detection kit, a preparation method and applications thereof. The test kit comprises: a test paper strip, wherein the test paper strip comprises a filtration pad, a sample pad, a detection pad and the like, a detection line and a quality control line are not coincident with each other, are distributed on the detection pad, and respectively contain a first antibody and a second antibody; and immunized nanometer silver gold core shell particles, wherein the immunized nanometer silver gold core shell particles contain functional polymer grafted nanometer silver gold core shell nanoparticles and a third antibody covalently bonded to a functional polymer, wherein the first antibody, a target and the third antibody are specifically bonded in a sandwich manner only in the presence of the target, and the second antibody and the third antibody are bonded in the presence or absence of the target. With the kit and the detection method of the present invention, the detection range and the detection sensitivity of the common food-borne pathogenic bacteria (especially Vibrio parahaemolyticus) in the seafood product can be substantially increased, the operation is simple, the detection time is short, and the favorable way is provided for the large batch and rapid detection of the seafood product.

Description

technical field [0001] The invention relates to a kit, in particular to a kit applicable to the detection of food-borne pathogenic bacteria in seafood products, such as vibrio parahaemolyticus, its preparation method and application, and belongs to the field of microbial detection. Background technique [0002] Vibrio parahaemolyticus is one of the common food-borne pathogens, which mainly come from seafood such as shrimp, fish and shellfish. According to the report of the National Foodborne Disease Surveillance Network, food poisoning caused by Vibrio parahaemolyticus has been on the rise since 1998 in my country, surpassing Salmonella food poisoning and ranking first, becoming the most serious foodborne pathogen. Human consumption of seafood infected by this fungus can cause gastrointestinal diseases such as gastroenteritis and sepsis, and it is the primary pathogenic bacteria in food poisoning cases in some coastal areas of my country. Since the discovery of Vibrio parah...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/531
Inventor 黄又举陈涛赖卫华王景云
Owner NINGBO INST OF MATERIALS TECH & ENG CHINESE ACADEMY OF SCI
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