Seafood product detection kit and preparation method thereof

A technology of hydroxyvaleric acid and guanidine thiocyanate, which is applied in the field of microbial detection, can solve problems such as false positives, missed detection, and complex components, and achieve the effects of increasing coupling rate, avoiding cross-linking and polymerization, and high sensitivity

Active Publication Date: 2020-07-10
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When the concentration of Vibrio parahaemolyticus in food is low, it is easy to miss detection by using conventional detection methods, while for more sensitive rapid detection methods such as ELISA or PCR, due to the relatively complex composition of food samples, false positives are prone to occur

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] 1. Preparation of Vibrio parahaemolyticus rabbit clone antibody IgG: the concentration is 1×10 9 CFU mL -1 The inactivated bacteria solution was emulsified completely with the same amount of Freund's complete adjuvant / Freund's incomplete adjuvant to prepare the inactivated vaccine. The concentration for the first immunization was 1×10 9 CFU mL -1 The healthy New Zealand white rabbits were immunized with Freund's complete adjuvant vaccine, and each rabbit was immunized with 2 mL by multi-point subcutaneous injection on the back. Immunization, the third immunization. After 10 days, the immunization was boosted twice with inactivated bacterial solution. Before each immunization, blood was collected from the vein of the rabbit's ear, and the serum titer was determined. When the separation standard is reached, blood is collected from the heart of the rabbit, and the serum is separated to obtain antiserum against Vibrio parahaemolyticus. The polyclonal antibody IgG in t...

Embodiment 2

[0063] Surface carboxyl functional modification of magnetic nanocomposite particles: Weigh 0.086g SDS sodium dodecyl sulfate and 0.008g NaHCO 3 In the three-necked flask, add 28mL of water and the prepared Fe 3 o 4 @SiO 2 The magnetic nanocomposite particles are ultrasonically dispersed and placed in a water bath at 72°C, stirred at 300 rpm, and protected by nitrogen gas. When the temperature of the liquid in the three-neck flask reaches 72°C, add 2 mL of styrene, 200 μL of 2-hydroxyvaleric acid (1.3 g / mL), and 0.5 mL of potassium persulfate (0.03 g / mL) to react for 8 hours. The magnetic suction pouring method washes the precipitate 5 times with ethanol, then washes it 5 times with ultrapure water, and finally dissolves the precipitate in ultrapure water to obtain carboxyl functionalized magnetic beads. All the other parts are completely consistent with Example 1.

Embodiment 3

[0065] Surface carboxyl functional modification of magnetic nanocomposite particles: Weigh 0.086g SDS sodium dodecyl sulfate and 0.008g NaHCO 3 In the three-necked flask, add 28mL of water and the prepared Fe 3 o 4 @SiO 2 The magnetic nanocomposite particles are ultrasonically dispersed and placed in a water bath at 72°C, stirred at 300 rpm, and protected by nitrogen gas. When the temperature of the liquid in the three-necked bottle reaches 72°C, add 2 mL of styrene, 200 μL of acrylic acid, and 0.5 mL of potassium persulfate (0.03 g / mL) to react for 8 hours. After the reaction is completed, wash the precipitate with ethanol for 5 times by magnetic suction pouring method , and then washed five times with ultrapure water, and finally dissolved the precipitate in ultrapure water to obtain carboxyl functionalized magnetic beads.

[0066] Preparation of anti-Vibrio parahaemolyticus immune magnetic beads: After the 1mg / mL carboxyl-functionalized magnetic beads are fully ultrasoni...

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Abstract

The invention provides a seafood product detection kit and a preparation method thereof, and belongs to the field of microbiological detection. The seafood product detection kit comprises anti-vibrioparahaemolyticus immunomagnetic beads and CdSe/ZnS-carboxylated polystyrene immunofluorescent microspheres. The magnetic beads provided by the invention are relatively high in surface carboxyl modification amount, the coupling rate of antibodies can be increased, and the capture rate of vibrio parahaemolyticus can be increased. According to the preparation method of the CdSe/ZnS-carboxylated polystyrene immunofluorescent microspheres, the cross-linking polymerization of the fluorescent microspheres is reduced, the dispersity is improved, the coupling rate of an antibody is improved, and the fluorescence intensity value is increased. The seafood product detection kit provided by the invention is used for detecting the vibrio parahaemolyticus and has relatively high sensitivity, accuracy andreliability.

Description

technical field [0001] The invention belongs to the field of microorganism detection, and in particular relates to a seafood product detection kit and a preparation method thereof. Background technique [0002] Vibrio parahaemolyticus (Vibrio parahaemolyticus) is an important foodborne pathogen, and the outbreak of foodborne disease caused by it is one of the most important public health problems. In order to effectively control Vibrio parahaemolyticus, it is very important to establish an efficient and sensitive separation and detection method. When the concentration of Vibrio parahaemolyticus in food is low, it is easy to miss detection by using conventional detection methods, while for more sensitive rapid detection methods such as ELISA or PCR, due to the relatively complex composition of food samples, false positives are prone to occur. Therefore, it is urgent to establish a method to make up for the above deficiencies. Immunomagnetic bead separation technology (Immun...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543G01N33/533G01N33/545
CPCG01N33/56911G01N33/54326G01N33/533
Inventor 应晓国施佩影相兴伟惠国华
Owner ZHEJIANG OCEAN UNIV
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