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Method for rapidly detecting salmonella

A Salmonella, rapid technology, applied in the field of microbial detection, can solve the problems of limited colloidal gold optical signal, limited application, low sensitivity of colloidal gold immunochromatographic test strips, etc., to improve detection sensitivity, reduce workload, and immunological reactions. uniform effect

Active Publication Date: 2016-05-04
NANCHANG UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

[0004] Colloidal gold immunochromatographic test strips have become an important tool for primary screening due to their simple operation, fast (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 Salmonella is usually not higher than 10 5 CFU / mL, this defect limits its application in food and agricultural product testing

Method used

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  • Method for rapidly detecting salmonella

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Embodiment 1

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

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

[0041] 1.1Fe 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 100mL of deionized water, nitrogen gas was passed into the solution and heated to 90°C, and then 4.7mL 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 silica ...

Embodiment 2

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

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

[0053] 1.1Fe 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 100mL of deionized water, nitrogen gas was passed into the solution and heated to 90°C, and then 4.7mL 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 silica ...

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Abstract

The invention provides a detection method for rapidly detecting salmonella. According to the scheme, bacteria are enriched by Fe3O4 / Ru(bqy)<2+>3 microspheres, a test strip is prepared, and a sample is loaded for detection. The step of eluting the salmonella from immunomagnetic beads is eliminated, so that the capturing efficiency is improved; the step of spraying the Fe3O4 / Ru(bqy)<2+>3 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 Salmonella by nano-microsphere integrated immunomagnetic bead capture technology and immunochromatography. technical background [0002] With the continuous improvement of people's living standards, food safety issues have been widely concerned and valued by all walks of life. Among the global food safety poisoning incidents, poisoning incidents caused by foodborne pathogens have caused serious losses and harm to human health and social development. Salmonella is a kind of food-borne pathogenic bacteria ubiquitous in nature, and the poisoning cases caused by Salmonella occupy the first place in food poisoning all over the world. Salmonella has a wide range of hosts in nature, and it is easy to spread directly or indirectly between animals, between animals and humans, and between humans. The main route of human infection with Sal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569G01N33/558G01N33/543
CPCG01N33/54346G01N33/558G01N33/56916G01N33/577
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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