Platform rapid microorganism detection method based on quaternary ammonium salt magnetic nanometer material and urease composite sensor

A composite sensor, quaternary ammonium salting technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of complex operation, extremely high operator requirements, and long time.

Inactive Publication Date: 2018-01-16
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Conventional detection methods are complicated to operate, time-consuming, and have extremely high requirements for operators, which are far from satisfying the curr...

Method used

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  • Platform rapid microorganism detection method based on quaternary ammonium salt magnetic nanometer material and urease composite sensor
  • Platform rapid microorganism detection method based on quaternary ammonium salt magnetic nanometer material and urease composite sensor
  • Platform rapid microorganism detection method based on quaternary ammonium salt magnetic nanometer material and urease composite sensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Detection of Staphylococcus aureus.

[0028] The microorganisms used in the experiment were cultured in suspension in lysed broth (1% peptone, 1% sodium chloride, 0.5% yeast extract, 100 mL water), and a single colony was cultured overnight at 30°C and 200 rpm on a shaking table at 4500 rpm. / Centrifuge for ten minutes, and dilute to different concentrations with PBS buffer solution.

[0029] 100 nM urease solution was incubated with 6nM q-MNP1 for 15min, and 100 µL concentration (10 cfu ml -1 ) bacterial solution and the above-mentioned nanomaterial and enzyme complex, reacted at 37 °C for 30min. Use a magnet to remove magnetic particles or magnetic particle-microbe complexes, then add 200 mM NaCl, 60 mM MgCl 2 , 200 microliters of 50mM urea and 0.4% phenol red solution, incubate for 30 minutes, and quickly detect the microbial intensity signal with a multi-functional microplate reader.

[0030] 100 nM urease solution was incubated with 6nM q-MNP1 for 15...

Embodiment 2

[0038] Example 2: Detection of Escherichia coli.

[0039] The microorganisms used in the experiment were cultured in suspension in lysed broth (1% peptone, 1% sodium chloride, 0.5% yeast extract, 100 mL water), and a single colony was cultured overnight at 30°C and 200 rpm on a shaking table at 4500 rpm. / Centrifuge for ten minutes, and dilute to different concentrations with PBS buffer solution.

[0040] 100 nM urease solution was incubated with 6nM q-MNP2 for 15min, and 100 µL concentration (10 cfu ml -1 ) bacterial solution and the above-mentioned nanomaterial and enzyme complex, reacted at 37 °C for 30min. Use a magnet to remove magnetic particles or magnetic particle-microbe complexes, then add 200 mM NaCl, 60 mM MgCl 2 , 200 microliters of 50mM urea and 0.4% phenol red solution, incubate for 30 minutes, and quickly detect the microbial intensity signal with a multi-functional microplate reader.

[0041] 100 nM urease solution was incubated with 6nM q-MNP2 for 15min, ...

Embodiment 3

[0049] Example 3: Detection of Edwardsiella tarda bacteria.

[0050] The microorganisms used in the experiment were cultured in suspension in lysed broth (1% peptone, 1% sodium chloride, 0.5% yeast extract, 100 mL water), and a single colony was cultured overnight at 30°C and 200 rpm on a shaking table at 4500 rpm. / Centrifuge for ten minutes, and dilute to different concentrations with PBS buffer solution.

[0051] 100 nM urease solution was incubated with 6nM q-MNP3 for 15min, and 100 µL concentration (10 cfu ml -1 ) bacterial solution and the above-mentioned nanomaterial and enzyme complex, reacted at 37 °C for 30min. Use a magnet to remove magnetic particles or magnetic particle-microbe complexes, then add 200 mM NaCl, 60 mM MgCl 2 , 200 microliters of 50mM urea and 0.4% phenol red solution, incubate for 30 minutes, and quickly detect the microbial intensity signal with a multi-functional microplate reader.

[0052] 100 nM urease solution was incubated with 6nM q-MNP3 ...

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Abstract

Marine pathogenic microorganism is an important factor capable of influencing national economical construction, so that pathogenic microorganism detection and control study possesses significant importance on national economical construction. The invention discloses a platform used for rapid direct analysis of microorganisms in environmental microorganism samples based on a quaternary ammonium salt magnetic nanometer material-urease composite sensor. According to the invention, study on recognition mechanisms and reaction kinetics of the quaternary ammonium salt magnetic nanometer material with a microorganism response material is carried out, study on the functional rules of identification response function modes in microorganism rapid detection and microorganism instant expression analysis is carried out. The invention discloses the action relationship of the quaternary ammonium salt magnetic nanometer material-urease composite sensor with microorganisms, conclusion of action rules of a one-stop microbial analyzer with different microorganisms, and provides reference for solving portable and automatic problems in microorganism detection method.

Description

technical field [0001] The invention designs a rapid detection of microorganisms based on a quaternary ammonium salt magnetic nanometer material and urease composite sensor platform. Background technique [0002] In the marine environment, pathogenic microbial pollution, water eutrophication, microbial corrosion, and microbial fouling are all apparent forms of microorganisms that threaten human production and life, and are also objective conditions for rapid microbial detection technology. Existing data show that the loss of pathogenic microbial contamination is closely related to the speed of microbial identification, the longer the identification time, the greater the loss. This is because, on the one hand, the rapid growth and spread of microorganisms aggravates environmental pollution and human diseases; on the other hand, the inability to identify the types of microorganisms makes it impossible to implement targeted protection programs, which in turn leads to the abuse ...

Claims

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Application Information

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IPC IPC(8): C12Q1/26C12Q1/14C12Q1/10C12Q1/04
Inventor 万逸方琳怡宋凤阁
Owner HAINAN UNIVERSITY
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