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A method for rapid detection of Vibrio parahaemolyticus

A hemolytic vibrio, rapid technology, applied in the field of microbial detection, can solve the problems of limited application, limited optical signal of colloidal gold, low sensitivity of colloidal gold immunochromatographic test strips, etc., and achieves improved detection sensitivity and good detection stability. , the effect of uniform immunological response

Active Publication Date: 2017-12-15
NANCHANG UNIV
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Problems solved by technology

[0004] Colloidal gold immunochromatographic test strips have become an important tool for basic screening due to their simple operation, rapidity (10-15min), and accuracy. However, due to the limited optical signal of colloidal gold, the sensitivity of colloidal gold immunochromatographic test strips is not high High, the limit of detection for Vibrio parahaemolyticus is usually not higher than 10 5 CFU / mL, this defect limits its application in food and agricultural product testing

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  • A method for rapid detection of Vibrio parahaemolyticus

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Embodiment 1

[0039] Embodiment one: using Fe 3 o 4 / Ru(bqy) 3 2+ Detection of Vibrio parahaemolyticus in milk by nano-microsphere immunochromatographic test strip

[0040] 1. Preparation of Fc-conjugated mAbs 3 o 4 / Ru(bqy) 3 2+ Nanospheres

[0041] 1.1 Fe 3 o 4 / Ru(bqy) 3 2+ Preparation of nanospheres: adding 0.6mmolFeCl 3 ·6H 2 O and 0.3 mmol FeCl 2 4H 2 O into 100 mL of deionized water, bubble nitrogen into the solution and heat to 90°C, then add 4.7 mL of 25% NH 3 ·H 2 O was added to the mixture and reacted for 2h. Use a permanent magnet to separate the black solid matter from the reaction solution and wash it with high-purity water for 3 to 5 times to obtain Fe 3 o 4 Nanoparticles; take 12mg Fe 3 o 4 Nanoparticles were resuspended with a mixture of 3mL deionized water and 20mL ethanol, and 0.5mL of NH 4 OH solution, then dissolve 50uL tetraethyl orthosilicate in 50uL ethanol solution and add dropwise, react at room temperature for 12h, in Fe 3 o 4 A layer of si...

Embodiment 2

[0051] Embodiment two: using Fe 3 o 4 / Ru(bqy) 3 2+ Detection of Vibrio parahaemolyticus in beef by nano-microsphere immunochromatographic test strip

[0052] 1. Preparation of Fc-conjugated mAbs 3 o 4 / Ru(bqy) 3 2+ Nanospheres

[0053] 1.1 Fe 3 o 4 / Ru(bqy) 3 2+ Preparation of nanospheres: add 0.6mmol FeCl 3 ·6H 2 O and 0.3 mmol FeCl 2 4H 2 O into 100 mL of deionized water, bubble nitrogen into the solution and heat to 90°C, then add 4.7 mL of 25% NH 3 ·H 2 O was added to the mixture and reacted for 2h. Use a permanent magnet to separate the black solid matter from the reaction solution and wash it with high-purity water for 3 to 5 times to obtain Fe 3 o 4 Nanoparticles; take 12mg Fe 3 o 4 Nanoparticles were resuspended with a mixture of 3mL deionized water and 20mL ethanol, and 0.5mL of NH 4 OH solution, then dissolve 50uL tetraethyl orthosilicate in 50uL ethanol solution and add dropwise, react at room temperature for 12h, in Fe 3 o 4 A layer of sili...

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Abstract

The invention provides a detection method for rapidly detecting vibrio parahaemolyticus. According to the scheme, bacteria are enriched by Fe3O4 / Ru(bqy) 3<2+> microspheres, a test strip is prepared, and a sample is loaded for detection. The step of eluting the vibrio parahaemolyticus from immunomagnetic beads is eliminated, so that the capturing efficiency is improved; the step of spraying the Fe3O4 / Ru(bqy) 3<2+> microspheres onto a conjugate pad is eliminated, so that immunological reaction is more uniform, and a variation coefficient during quantitative detection is small; the workload and the sundry fungus contamination probability are reduced; the detection scheme is high in sensitivity and stability.

Description

technical field [0001] The invention relates to the field of microbial detection, specifically using Fe 3 o 4 / Ru(bqy) 3 2+ Rapid detection of Vibrio parahaemolyticus using nanosphere integrated immunomagnetic bead capture technology and immunochromatography. technical background [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. [0003] Since the discovery of Vibrio parahaemolyticu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/569G01N33/558G01N33/543
CPCG01N33/54346G01N33/558G01N33/56911G01N33/577
Inventor 赖卫华王景云刘道峰邓省亮山珊熊勇华魏华
Owner NANCHANG UNIV
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