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Method for quickly detecting pathogenic microorganism through multicolor fluorescence magnetic chemical nose sensor

A sensor and blue fluorescent protein technology, applied in the field of biosensing, can solve the problems of difficult analysis results due to technical requirements, cumbersome microbial antibody screening, low detection specificity, etc., to shorten the analysis time, facilitate the separation of target bacteria, The effect of simplifying the analysis and detection steps

Inactive Publication Date: 2019-01-01
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional detection method is mainly based on the physiological and biochemical characteristics of the bacteria. After the steps of pre-enrichment, selective plate separation, and biochemical identification, it takes 3-5 days from sampling to confirming the results. The detection cycle is long, the operation is cumbersome, and the workload is heavy.
Using the specificity of antigen-antibody reaction to identify bacteria has a history of more than half a century, but the screening of microbial antibodies is very cumbersome, and the final detection specificity is not high
Of course, with the continuous improvement and development of molecular biology detection technology, the rapid detection method for pathogenic microorganisms using PCR technology has been developed rapidly, but the samples need special treatment, have high technical requirements and are not easy to analyze the results

Method used

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  • Method for quickly detecting pathogenic microorganism through multicolor fluorescence magnetic chemical nose sensor
  • Method for quickly detecting pathogenic microorganism through multicolor fluorescence magnetic chemical nose sensor
  • Method for quickly detecting pathogenic microorganism through multicolor fluorescence magnetic chemical nose sensor

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Experimental program
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Effect test

Embodiment 1

[0036] "Chemical nose" sensor construction: the concentration of (100 n M) blue fluorescent protein, green fluorescent protein and red fluorescent protein and phenyl functional group quaternary ammonium salted magnetic nanoparticles are titrated until three kinds of fluorescence intensity Reduce to the lowest value. The fluorescent polymer-magnetic "chemical nose" sensor can be obtained.

Embodiment 2

[0038] Salmonella typhi was cultured in LB medium at 37°C for 12 hours, then washed three times with PBS and diluted to a concentration of 5×10 8 cfuml -1 , Take 100μL of the above bacterial liquid and 100μL of nanosensor complex and incubate for 15min. A magnet is used to remove magnetic particles or magnetic particle-microbe complexes, and a microplate reader is used to measure the response signals of the three fluorescent arrays. Combining with linear discriminant analysis can determine the species of Salmonella typhi.

[0039] Salmonella typhi was cultured in LB medium at 37°C for 12 hours, then washed three times with PBS and diluted to a concentration of 5×10 6 cfu ml -1 , Take 100μL of the above bacterial liquid and 100μL of nanosensor complex and incubate for 15min. A magnet is used to remove magnetic particles or magnetic particle-microbe complexes, and a microplate reader is used to measure the response signals of the three fluorescent arrays. Combining linear discrim...

Embodiment 3

[0042] Enterococcus faecalis was cultured in LB medium at 37°C for 12 hours, then washed three times with PBS and diluted to a concentration of 5×10 8 cfuml -1 , Take 100μL of the above bacterial liquid and 100μL of nanosensor complex and incubate for 15min. A magnet is used to remove magnetic particles or magnetic particle-microbe complexes, and a microplate reader is used to measure the response signals of the three fluorescent arrays. Combined with linear discriminant analysis, the species of Enterococcus faecalis can be determined.

[0043] Enterococcus faecalis was cultured in LB medium at 37°C for 12 hours, then washed three times with PBS and diluted to a concentration of 5×10 6 cfu ml -1 , Take 100μL of the above bacterial liquid and 100μL of nanosensor complex and incubate for 15min. A magnet is used to remove magnetic particles or magnetic particle-microbe complexes, and a microplate reader is used to measure the response signals of the three fluorescent arrays. Combi...

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Abstract

The invention belongs to the field of microorganism detection, and particularly discloses a method for quickly detecting a pathogenic microorganism through a multicolor fluorescence magnetic chemicalnose sensor and screening antibiotics acting on the microorganism. The chemical nose sensor comprises multicolor fluorescence protein and hyamine magnetic nanoparticles. Based on the chemical nose sensor, signals of the multicolor fluorescence protein are quenched, and then the the chemical nose sensor is competitively combined with a to-be-detected pathogenic microorganism, the multicolor fluorescence protein freely gives out a fluorescence signal, and response signals of different microorganisms are obtained according to the variation difference of the fluorescence signals; and furthermore,the chemical nose sensor is competitively combined with the to-be-detected microorganism which is subjected to action of the antibiotics to screen the antibiotic. The chemical nose sensor is simple tooperate, convenient to use, high in sensitivity and high in stability, can be used for rapidly detecting and screening the pathogenic microorganism and the antibiotic acted to the pathogenic microorganism, and can provide reference in the aspects of microorganism infection, antibiotic identification, disease diagnosis and the like.

Description

Technical field [0001] The invention belongs to the technical field of biosensing, and specifically relates to a multicolor fluorescent magnetic chemical nose sensor that rapidly detects pathogenic microorganisms and responds to microorganisms that have undergone the action of antibiotics, thereby realizing screening of antibiotics. Background technique [0002] Microorganisms are a large group of organisms, including bacteria, viruses, fungi, and some small protozoa. They are tiny and widely distributed. Their life activities are closely related to human daily life and production. Food, health, medicine, industry and agriculture, environmental protection, energy development and many other fields are inseparable from the contribution of microorganisms. However, in addition to microorganisms that are beneficial to humans, there are some microorganisms in nature that can cause pathological damage to humans, animals and plants. Such microorganisms are called "pathogenic microorgani...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 万逸方琳怡宋凤阁
Owner HAINAN UNIVERSITY
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