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A method for rapid detection of Escherichia coli o157:h7

A technology of Escherichia coli and O157, applied in the field of microbial detection, can solve the problems of limited application, limited colloidal gold optical signal, and low sensitivity of colloidal gold immunochromatographic test strips, and achieves improved detection sensitivity, good detection stability, and immune uniform effect

Active Publication Date: 2018-01-16
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 E. coli O157:H7 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 Escherichia coli o157:h7

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Embodiment 1

[0039] Embodiment one: using Fe 3 o 4 / Ru(bqy) 3 2+ Detection of Escherichia coli O157:H7 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 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 s...

Embodiment 2

[0051] Embodiment two: using Fe 3 o 4 / Ru(bqy) 3 2+ Detection of Escherichia coli O157:H7 in beef with 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: 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 s...

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Abstract

The invention provides a detection method for fast detecting colon bacillus O157:H7. According to the scheme, Fe3O4 / Ru(bqy)3<2+> nanoparticles are subjected to bacterium enriching; a test strip is prepared; sample loading detection is performed. The step of eluting the colon bacillus O157:H7 from immunomagnetic beads is omitted, so that the capture efficiency is improved; the step of spraying Ru(bqy)3<2+> nanoparticles onto a combining pad is omitted, so that the immunological reaction is more uniform, and the variable coefficient is small during the quantitative determination; the workload and the infectious microbe contamination probability are reduced. The sensitivity of the detection scheme is very high, and the stability is high.

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 Escherichia coli O157:H7 by nanosphere integrated immunomagnetic bead capture technology and immunochromatography. technical background [0002] In recent years, with the improvement of people's living standards, food safety has become a global public health hotspot. Food-borne pathogens are a major hidden danger to food safety. Escherichia coli O157: H7 is currently recognized as one of the three most important food-borne pathogens in the world. Its infection dose is very low, at least 10 live Bacteria can cause disease. The clinical symptoms of Escherichia coli O157:H7 infection include diarrhea, hemorrhagic colitis, hemolytic uremic syndrome, and thrombocytopenic purpura, etc. The average period from exposure to the bacteria to onset is 3 days, and the incubation period is as short as 1 day. The longest can reach more than 8 ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/558
CPCG01N33/558G01N33/56916
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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