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PCR (Photo-conductive Relay) and capillary electrophoresis integrated micro-fluidic chip and rapid pathogenic bacteria detection device adopting chip

A technology of microfluidic chip and capillary electrophoresis, which is applied in enzymology/microbiology devices, biochemical cleaning devices, biochemical equipment and methods, etc. It can solve the problem of increasing operational errors and sample contamination, and it is difficult to reduce reagent and labor costs. The cumbersome operation process and other problems can be achieved to reduce the risk of sample contamination, reduce hardware purchase costs, and simplify manual operations.

Inactive Publication Date: 2015-07-01
窦晓鸣 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This puts forward relatively high requirements on the hardware conditions of the laboratory and the operation level of the experimenters, and the operation process is relatively cumbersome, which makes it difficult to reduce the cost of reagents and labor in the detection process.
Sample transfer between multiple devices also increases the possibility of operator error and sample contamination

Method used

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  • PCR (Photo-conductive Relay) and capillary electrophoresis integrated micro-fluidic chip and rapid pathogenic bacteria detection device adopting chip
  • PCR (Photo-conductive Relay) and capillary electrophoresis integrated micro-fluidic chip and rapid pathogenic bacteria detection device adopting chip
  • PCR (Photo-conductive Relay) and capillary electrophoresis integrated micro-fluidic chip and rapid pathogenic bacteria detection device adopting chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1. A microfluidic chip integrated with PCR and capillary electrophoresis

[0043] The microfluidic chip integrated with PCR and capillary electrophoresis is 76mm long, 22mm wide, and 2mm thick. It is formed by bonding and sealing the upper and lower layers of PDMS substrates. Both the thickness of the upper and lower substrates are 1 mm. The lower substrate contains PCR reaction channels, CE separation channels and groove structures. The upper substrate contains hole structures whose positions correspond to the groove structures of the lower substrate.

[0044] Wherein, the structure of the lower substrate 7 is as follows figure 2 shown. The cross-sectional size of the PCR channel 8 is 0.1mm×0.1mm, and every two adjacent linear parts form a group, which can complete one PCR reaction, and there are 30 groups in total. The CE channel 12 has a cross-sectional dimension of 0.1 mm×0.1 mm and a total length of 16 mm. The linear part at the end of the PCR channel c...

Embodiment 2

[0046] Example 2. Rapid detection of Porphyromonas Gingivalis (Pg) using a microfluidic chip and a detection device.

[0047] The gingival sulcus was wiped with a hygroscopic paper tip (Tianjin Dayading Medical Instrument Co., Ltd.) to obtain gingival crevicular fluid containing oral bacteria. Soak the tip part of the absorbent paper tip stained with gingival crevice fluid in the lysing solution PBS (Sigma-Aldrich, USA) for 3 minutes. PCR channel 8 of the microfluidic chip was filled with PCR reaction solution SpeedSTAR HS DNA Polymerase (TAKARA, Japan), and CE channel 12 was filled with separation solution (hydroxyethyl cellulose solution). Insert the chip into the detection device, inject 1 μL of lysate containing oral bacterial DNA into the chip injection pool 15, turn on the device, and automatically complete the subsequent steps. Under the order of the data processing and control module 1, the temperature control module 5 makes the temperature control sheet 13 reach 95°C...

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Abstract

The invention relates to a PCR (Photo-conductive Relay) and capillary electrophoresis integrated micro-fluidic chip and a rapid pathogenic bacteria detection device adopting the chip. The rapid pathogenic bacteria detection device adopting the chip comprises an upper baseplate and a lower baseplate, wherein the upper baseplate and the lower baseplate are bonded together; a PCR reaction channel and a CE (capillary electrophoresis) separation channel are formed in the lower baseplate; at least one cross point exists in the PCR reaction channel and the CE seperation channel, and a groove is respectively formed in each of ports of the PCR reaction channel and the CE seperation channel; and holes for filling a reagent and holding an electrode are formed in parts, opposite to the grooves, of the upper baseplate. Compared with a conventional gene analysis process, a method of detecting bacteria by applying the PCR and CE integrated micro-fluidic chip and the detection device has the advantages that the device quantity is reduced, the acquisition cost of hardware is reduced, and the degree of automation of an analysis process is increased.

Description

technical field [0001] The invention relates to the technical field of molecular biological detection, in particular to the detection of pathogenic bacteria. Background technique [0002] The detection and identification of organisms or tissues using specific gene sequences (fragments) as detection targets has become an important technical means in the fields of biology, biochemistry, medicine, environment and food, and has been widely used. Routine genetic analysis processes usually require the use of multiple devices such as centrifuges, temperature cyclers, slab gel electrophoresis, fluorescence imaging analyzers, or capillary electrophoresis. This puts forward relatively high requirements on the hardware conditions of the laboratory and the operation level of the experimenters, and the operation process is relatively cumbersome, which makes it difficult to reduce the cost of reagents and labor during the detection process. Sample transfers between multiple devices also ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12M1/38C12M1/36C12M1/34
Inventor 山口佳则倪一李振庆窦晓鸣
Owner 窦晓鸣
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