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Method for rapidly detecting staphylococcus aureus

A staphylococcus and golden yellow technology, applied in the field of microbial detection, can solve the problems of limited optical signal of colloidal gold, low sensitivity of colloidal gold immunochromatography test strips, and limited application, so as to improve detection sensitivity and uniform immunological response , detection of good stability

Active Publication Date: 2016-06-08
江西维邦生物科技有限公司
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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 Staphylococcus aureus 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 rapidly detecting staphylococcus aureus

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Embodiment 1

[0039] Embodiment one: using Fe 3 o 4 / Ru(bqy) 3 2+ Detection of Staphylococcus aureus 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...

Embodiment 2

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

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Abstract

The invention provides a method for rapidly detecting staphylococcus aureus. According to the scheme, the method comprises the steps of enriching Fe3O4 / Ru (bqy) 3<2+> nanoparticles with bacteria, preparing test strips, and mounting a sample for detection. By means of the method, the step of eluting staphylococcus aureus away from immunomagnetic beads is omitted, and capturing efficiency is improved; the step of spraying Ru (bqy) 3<2+> nanoparticles to a binding pad is omitted, an immunology reaction is more uniform, and the variation coefficient generated in the quantitative detection process is small; working load is reduced, and the probability of sundry bacteria contamination is lowered. The detection scheme is quite high in sensitivity and quite good in 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 Staphylococcus aureus by integrating immunomagnetic bead capture technology and immunochromatography with nano-microspheres. technical background [0002] Staphylococcus aureus is one of the most common food poisoning pathogens and can be found everywhere in nature. Therefore, there is a high chance that food will be contaminated by it. According to the report of the Centers for Disease Control and Prevention in the United States, food poisoning caused by Staphylococcus aureus is second only to Escherichia coli, accounting for 33% of the total bacterial food poisoning, and up to 45% in Canada. Staphylococcus aureus is one of the main pathogenic bacteria that cause clinical infection and surgical incision infection. It can cause local suppurative infection, pneumonia, pseudomembranous enteritis, pericarditis, etc., and even system...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/577
CPCG01N33/558G01N33/577
Inventor 赖卫华王景云刘道峰邓省亮山珊熊勇华魏华
Owner 江西维邦生物科技有限公司
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