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Method for fast detecting Listeria monocytogenes

A fast technology for Listeria monocytogenes, applied in the field of microbial detection, can solve the problems of limited colloidal gold optical signal, limited application, low sensitivity of colloidal gold immunochromatography test strips, etc., to improve detection sensitivity and reduce workload , Uniform immunological response

Active Publication Date: 2016-04-27
NANCHANG UNIV
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  • Abstract
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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 Listeria monocytogenes is usually not higher than 10 5 CFU / mL, this defect limits its application in food and agricultural product testing

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  • Method for fast detecting Listeria monocytogenes

Examples

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Embodiment 1

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

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

[0041] 1.1 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 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 lay...

Embodiment 2

[0051] Embodiment two: using Fe 3 o 4 / Ru(bqy) 3 2+ Detection of Listeria monocytogenes 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 laye...

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Abstract

The invention provides a detection method for fast detecting Listeria monocytogenes. 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 Listeria monocytogenes 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 Listeria monocytogenes by nanosphere integrated immunomagnetic bead capture technology and immunochromatography. technical background [0002] Listeria monocytogenes, referred to as Listeria monocytogenes for short, is a foodborne pathogen that is zoonotic. The bacterium is widely distributed in nature and can still grow and reproduce in food stored in a refrigerator at 4°C. It is one of the main pathogens in refrigerated food that threaten human health. Listeria monocytogenes can easily contaminate meat, eggs, seafood, dairy products, vegetables and other foods, causing food poisoning and listeriosis. Listeriosis has the characteristics of frequent infections during pregnancy, severe fetal complications, susceptibility to immunocompromised populations, high incidence of meningoencephalitis, and high mortality, which seriously th...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/558
CPCG01N33/558G01N33/56911
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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