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Micro-fluidic chip, detection system based on micro-fluidic chip and detection method of bacteria

A microfluidic chip and bacteria technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of complex signal detection instruments, poor reproducibility of electrochemical methods, low sensitivity of colorimetric methods and fluorescence methods, etc. Strong light, low reagent consumption, good biocompatibility

Active Publication Date: 2022-03-29
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Each of these methods has certain shortcomings, among which the sensitivity of the colorimetric method and the fluorescence method is not high, the reproducibility of the electrochemical method is poor, and the surface Raman scattering method and the surface plasmon resonance method require complex and expensive signal detection instruments

Method used

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  • Micro-fluidic chip, detection system based on micro-fluidic chip and detection method of bacteria
  • Micro-fluidic chip, detection system based on micro-fluidic chip and detection method of bacteria
  • Micro-fluidic chip, detection system based on micro-fluidic chip and detection method of bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1: Structure of microfluidic chip

[0066] This embodiment provides a microfluidic chip, the three-dimensional structure of which is as follows: figure 1 shown. The microfluidic chip is composed of a bonded substrate and a cover sheet. In this embodiment, the material of the substrate is glass, and the material of the cover is polydimethylsiloxane (PDMS). The cover sheet is provided with a serpentine mixing microchannel 1 and a boat-shaped reaction microchannel 2, and the micropillar arrays 3 are evenly distributed in the boat-shaped reaction microchannel 2. The boat-shaped reaction microchannel 2 includes two bows and a hull between the two bows, one bow is used as an inlet, and the other is used as an outlet. The mixing microchannel includes 2 inlets and 1 outlet, and the 2 inlets are used as the inlets of luminol and hydrogen peroxide respectively. The outlet of the mixing microchannel is connected to the bow inlet of the boat-shaped reaction microchannel...

Embodiment 2

[0071] Example 2: Preparation of microfluidic chip

[0072] The silicon wafer was firstly ultrasonicated with ethanol for 5 minutes, and then deionized water for 5 minutes, and then heated on a heating plate for 120 minutes after performing three times of ultrasonication.

[0073] A thin layer of SU-82050 negative photoresist was spin-coated on the silicon wafer, and the thickness of the adhesive layer was about 50 μm by controlling the rotating speed. will have figure 2 The mask of the structure is bonded to the silicon wafer, the ultraviolet light passes through the photomask to expose the photoresist, and the unexposed part is dissolved with a developer. The raised SU-8 structure on the silicon wafer surface serves as a positive mold for the PDMS cover slip. The silicon positive mold was then silanized overnight.

[0074] The PDMS prepolymer (monomer / curing agent = 10 / 1 mixed) is poured on the silicon positive mold and vacuum degassed, then placed in a 75°C oven to cure...

Embodiment 3

[0080] Embodiment 3: detection system based on microfluidic chip

[0081] This embodiment provides a detection system based on a microfluidic chip, such as Figure 4 As shown, it includes the microfluidic chip d described in Example 1, a sampling unit and a detection and analysis unit, and the sampling unit communicates with the inlet of the mixing microchannel of the microfluidic chip.

[0082] In a specific embodiment, the detection and analysis unit includes a PMT detector f and an ultra-weak chemiluminescence analyzer g. The size of the microfluidic chip matches the card slot in the box of the PMT detector (25*25mm), and after it is placed, the boat-shaped reaction microchannel just matches the position of the light inlet of the PMT detector, and the chemical produced The luminescence signal was collected by PMT and analyzed by ultra-weak chemiluminescence analyzer g.

[0083] The sampling unit includes a micro-syringe pump a, a PTFE catheter c and a syringe b. The micro...

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Abstract

The invention discloses a micro-fluidic chip, a detection system based on the micro-fluidic chip and a detection method of bacteria, the micro-fluidic chip is composed of a substrate and a cover plate which are bonded together, the cover plate is provided with a mixing micro-channel and a reaction micro-channel, the mixing micro-channel and the reaction micro-channel are connected with each other, the reaction micro-channel is internally provided with a micro-column array, and the micro-column array is connected with the micro-column array. Furthermore, hairpin-shaped oligonucleotide 1 is modified on the microcolumn. According to the detection method, two hairpin-like oligonucleotides (H1 and H2) are triggered to generate a catalytic hairpin self-assembly reaction through competitive binding of the pathogenic bacteria, the reaction not only amplifies the concentration of the pathogenic bacteria, but also fixes horse radish peroxidase on the micro-column array, and further catalyzes luminol and hydrogen peroxide to generate a chemiluminescence reaction, so that the concentration of the pathogenic bacteria is detected. The pathogenic bacteria are quantified according to the chemical reaction signal intensity. The method has good specificity, low detection limit and wide linear range, and provides accurate data for quantitative detection of food-borne pathogenic bacteria.

Description

technical field [0001] The invention belongs to the technical field of microfluidic chips, in particular to a microfluidic chip, a detection system based on a microfluidic chip and a detection method for bacteria, in particular to a microfluidic chip for pathogenic bacteria, a microfluidic chip based Fluidic chip detection system and pathogenic bacteria detection method. Background technique [0002] Foodborne pathogens are pathogenic bacteria that can cause food poisoning or use food as a medium for transmission. Pathogenic bacteria directly or indirectly contaminate food and water sources, which can lead to infectious diseases of livestock and poultry or human intestinal infectious diseases and food poisoning, which is one of the root causes of food safety problems. Common foodborne pathogens mainly include pathogenic Escherichia coli, Salmonella, Staphylococcus aureus, and Listeria monocytogenes. Escherichia coli O157:H7 (E.coil O157:H7) is one of the main foodborne pat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543G01N21/76
CPCG01N33/56911G01N33/54346G01N21/76Y02A50/30
Inventor 蒋宇扬高丹孙冬丽樊婷婷
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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