Rapid bacterial detection method based on upconversion fluorescence resonance energy transfer

A fluorescence resonance energy and bacterial detection technology, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, measuring devices, etc. , high sensitivity and specificity

Active Publication Date: 2016-11-09
XI AN JIAOTONG UNIV +1
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Problems solved by technology

[0006] Therefore, from the overall level, although the traditional plate counting detection technology has low requirements, does not require special instruments, and is cheap, it is time-consuming and laborious, and the detection is not accurate
Although the combination of immunological rapid detection

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  • Rapid bacterial detection method based on upconversion fluorescence resonance energy transfer
  • Rapid bacterial detection method based on upconversion fluorescence resonance energy transfer
  • Rapid bacterial detection method based on upconversion fluorescence resonance energy transfer

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Embodiment Construction

[0028] The present invention will be further described in detail below in conjunction with specific embodiments, which are explanations of the present invention rather than limitations.

[0029] The rapid bacterial detection method based on upconversion fluorescence resonance energy transfer of the present invention comprises the following steps:

[0030] 1) NaOH, NH 4 F. YCl 3 and the chloride of the rare earth activator and the chloride of the rare earth sensitizer are added to the organic solvent, heated to 280-290° C., and kept for 0.5-3 hours to make the rare-earth doped up-conversion fluorescent nanoparticles; (the specific preparation process, You can refer to Chinese patent: ZL20131 0439588.6).

[0031] 2) Using the surface ligand exchange method, polyacrylic acid is used for water-soluble modification of rare earth-doped up-conversion fluorescent nanoparticles;

[0032] 3) Choose a biochemical method to synthesize the target bacterial aptamer and the cDNA single st...

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Abstract

The invention discloses a rapid bacterial detection method based on upconversion fluorescence resonance energy transfer, and belongs to the technical field of bacterial detection. The method includes the steps that the gold nanoparticle surface is modified with nucleic acid aptamers capable of specifically recognizing a target bacterium, the rare earth upconversion fluorescence particle surface is modified with cDNA paired with the aptamers complementarily, after the two kinds of nanoparticles are mixed, the distance between the two kinds of particles is reduced due to base complementary pairing of two nucleotide sequences, and therefore fluorescence of upconversion fluorescence particles is quenched by gold nanoparticles. When a target object is added, the target object and cDNA are competitively combined with the aptamers to recover fluorescence of upconversion fluorescence particles. The target bacterium is measured quantitatively according to different fluorescence recovery strength. The method is high in sensitivity and specificity, easy to operate, various target objects can be determined by changing aptamer sequences, and the rapid bacterial detection method is of great significance in environment monitoring and food analysis.

Description

technical field [0001] The invention belongs to the technical field of bacteria detection, and in particular relates to a high-sensitivity and high-specificity rapid bacteria detection method based on upconversion fluorescence resonance energy transfer. Background technique [0002] Foodborne pathogens are one of the most prevalent problems in the world today. On average, 1 out of 10 people in the world gets sick every year due to ingestion of pathogenic bacteria. [0003] E. coli is considered an indicator of fecal contamination. When E. coli is detected in water and food, it proves that there may be fecal contamination. Therefore, the number of coliform bacteria or the value of coliform bacteria is often used as the hygienic standard of drinking water and food. [0004] Escherichia coli is a resident bacterium in the intestinal tract of organisms, most of which are not pathogenic, but can cause extraintestinal infection under certain conditions. Pili strains of Escheri...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/06G01N33/569G01N33/533
CPCG01N33/533G01N33/56911C12Q1/6818C12Q2525/205C12Q2537/161C12Q2563/107C12Q2563/155
Inventor 林敏徐峰金碧瑞卢天健金瀛张淑静
Owner XI AN JIAOTONG UNIV
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