CN107841535A - A kind of application of micro-fluidic chip in microorganism detection - Google Patents
A kind of application of micro-fluidic chip in microorganism detection Download PDFInfo
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- CN107841535A CN107841535A CN201711353932.4A CN201711353932A CN107841535A CN 107841535 A CN107841535 A CN 107841535A CN 201711353932 A CN201711353932 A CN 201711353932A CN 107841535 A CN107841535 A CN 107841535A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
- C12Q1/6837—Enzymatic or biochemical coupling of nucleic acids to a solid phase using probe arrays or probe chips
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Abstract
The present invention proposes a kind of application of micro-fluidic chip in microorganism detection, micro-fluidic chip has convenient, efficiently, the features such as high flux, but micro-fluidic chip is due to integrated function, the degree of miniaturization is not so good as other chips, therefore the direction that more small chip can be studied as micro-fluidic chip from now on.
Description
Technical field
The present invention relates to field of biological detection, particularly relates to a kind of application of micro-fluidic chip in microorganism detection.
Background technology
Biochip (biochip) is the micro-analysis of a kind of high flux, parallel parsing that occur early 1990s
Technology.It builds and different lifes is secured on the carrier materials such as sheet glass, silicon chip, plastic sheet, nylon membrane or gel, on surface
Thing molecule is (such as：Oligonucleotides, cDNA, genomic DNA, polypeptide, antigen, antibody etc.), formation can be made mutually with purpose target molecule
With the solid phase surface with reaction.Chip and target biomolecule to be checked are chemically reacted (such as hybridization, immune response), then
The collection of signal is carried out using corresponding detection means, draws testing result.
Biochip with the advantages of high flux, high sensitivity, high specific to be widely used in cause of disease in fresh food
The detection of microorganism, its main feature have：(1) high flux：Probe is arranged in by filter using in-situ synthesis or direct point sample method
Micro-matrix is formed on the media such as film, silicon chip, glass, after measuring samples isotope or fluorescence molecule mark, with chip hybridization, is led to
Overscanning and computer analysis can obtain.Substantial amounts of gene order and expressing information in sample, to reach quick, efficient, high
Analyze the purpose of biological information flux；(2) high sensitivity：The sensitiveness of chip testing result is 96.2%, every kind of virus inspection
The minimum flow of survey is 10copies/ μ L, make biochip have compared with high sensitivity；(3) high specific：Chip is in hybridization reaction
After eluted, make not combined with target or the part of non-specific binding wash away, the specificity of chip testing result
For 99.3%.Played more in large biological molecule functional study, food safety detection, the pathogenic microorganism examination and identification etc.
Carry out more important effect.Several biological chip testing technologies and its application in microorganism detection have been sketched herein, for biology
Chip is in microorganism detection extensively using offer data reference.
Biochip constructs difference according to it can be divided into array cake core, micro-fluidic chip, nano chips etc.；According to its point
The probe difference of analysis can be divided into genetic chip, carbohydrate chip, protein chip, cell chip, organization chip etc.；It is different according to its application
Chip of expression spectrum, diagnosing chip, detection chip, genome snp analysis chip, genome chromosome analysis of variance chip can be divided into
And Pharmaceutical Analysis chip etc..The characteristics of various chips have its each, but all there is high flux, high specific and efficient spy
Point.
The great advantage of micro-fluidic chip is the mistake for being pre-processed, being reacted or being analyzed to sample in small space
Journey, so as to reach integrated function.On solid substrate build fluid channel, Micropump, micro-valve, reactor, blender, filter,
The tiny devices such as separator, realize preparation, biochemical reaction, liquid-phase chromatographic analysis, PCR reactions, the electrophoresis inspection of biological sample
The operation such as survey.Its target be by the function in whole laboratory, including sample, dilute, reagent adding, reaction, separation, the collection such as detection
Into on microchip.Micro-fluidic chip can be used for reacting or analyzing, and be a very important technology in biotechnology, can
The imagination of Microscale experiment room is implemented as, testing sample need to be only added directly into chip, chip will perform multiple automatically
Miscellaneous experiment flow, meanwhile, multiple samples or multiple reactions can be handled simultaneously in same chip, and can be tied rapidly
Fruit.Micro-fluidic chip, high flux and quickly analysis are carried out to protein, stability is higher, be a kind of integration, miniaturization,
It is easy to the detection and analysis device of automation.The characteristics of microfluidic system, is mainly manifested in high flux, low dosage, quick analysis and has
There is convenience etc., applied to DNA analysis, immunoassay and cell analysis.Micro-fluidic chip can be used for quick point
The biological property of microorganism is analysed, the detection to microorganism has highly important meaning.
The content of the invention
The present invention proposes a kind of application of micro-fluidic chip in microorganism detection.
The technical proposal of the invention is realized in this way：A kind of application of micro-fluidic chip in microorganism detection.
Further, including：One micro-fluidic chip carrys out detection bacterium DNA.Chip is by dimethyl silicone polymer (PDMS)
Come what is prepared, it includes SPE (SPE) room, two independent passages and multiloop mediated isothermality amplification (LAMP) room.
The design of microcavity and multiple microcavitys, meet multiple samples while detect have efficiently, saving sample, portable etc.
Feature, and successfully apply among the detection pathogenic bacteria such as salmonella, comma bacillus.Micro-fluidic chip is not only portable, and
And the sensitivity of detection is also higher, chip application is tested in the DNA hybridization of biological sample, using DNA specific probe hybridization skills
Art detects pathogen.According to base pair complementarity principle, single-stranded by known to " DNA probe " can be used as, then with other unknown
The solution mixing of DNA sequence dna can judge whether complementary DNA fragment by DNA probe, using the method for fluorescence labeling,
Under 532nm green glow, the presence of biological sample is detected, sensitivity is higher than conventional colorimetric method by 1 000~and 500 000 times, this is
A kind of sensitiveer, quick, accurate detection method.It is convenient that micro-fluidic chip has, efficiently, the features such as high flux, but it is micro-
For fluidic chip due to integrated function, the degree of miniaturization is not so good as other chips, thus more small chip can be used as it is micro- from now on
The direction of fluidic chip research.
Embodiment
Below in conjunction with the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described,
Obviously, described embodiment is only part of the embodiment of the present invention, rather than whole embodiments.Based in the present invention
Embodiment, the every other embodiment that those of ordinary skill in the art are obtained under the premise of creative work is not made, all
Belong to the scope of protection of the invention.
A kind of application of micro-fluidic chip in microorganism detection.
Further, including：One micro-fluidic chip carrys out detection bacterium DNA.Chip is by dimethyl silicone polymer (PDMS)
Come what is prepared, it includes SPE (SPE) room, two independent passages and multiloop mediated isothermality amplification (LAMP) room.
Three plants of bacterium (Escherichia coli O 157s:H7, methicillin-resistant staphylococcus aureus and methicillin-sensitivity are golden yellow
Staphylococcus) it is used to test the feasibility of the equipment.The yield of more ring mediated isothermal amplifications is directly detected by ultraviolet light, and by
Agarose gel electrophoresis confirms.Using this chip, these three bacterium, the detection limit of three kinds of bacterium are successfully detected within 2h
Less than 102CFU/100μL.But the chip also needs other equipment to confirm result, when real-time online detection can more shorten detection
Between, in the micro-fluidic chip system of a multichannel, fluoroscopic examination is carried out by light emitting diode, by CdSe/ZnS as fluorescence
Mark point, improve the sensitivity of detection.This micro-fluidic chip is by 12 sample channels, 3 mixed zones and 6 immune responses
District's groups are into using immune response, by fluoroscopic examination, detection limit is in 37CFU/mL.This fluorescence in situ in wire tag detects
Multichannel micro-fluidic chip be successfully applied to the detection of the salmonella typhimurium in pork sample.Micro-fluidic chip can
To be detected as a kind of microscale experiment room to microorganism, a variety of pathogens are detected using micro-fluidic chip.Pass through
Micro-fluidic chip carries out DNA/RNA extraction, the detection of real-time fluorescence PCR, by micro-fluidic chip be designed to one it is small-sized can
Mobile laboratory.This chip tool applies new optical detecting method, the design of microcavity and multiple microcavitys, meets multiple
Sample detects simultaneously, has the features such as efficient, saving sample, portable, and successfully apply in detection salmonella, cholera arc
Among the pathogenic bacteria such as bacterium.Micro-fluidic chip is not only portable, and the sensitivity detected is also higher, and chip application is in biological sample
DNA hybridization is tested, using DNA specific probe hybridization technology for detection pathogens., can be by known to according to base pair complementarity principle
It is single-stranded be used as " DNA probe ", then with other unknown DNA sequence dnas solution mixing can be judged whether by DNA probe
Complementary DNA fragment, using the method for fluorescence labeling, under 532nm green glow, detect the presence of biological sample, sensitivity than
Conventional colorimetric method is high by 1 000~and 500 000 times, this is a kind of sensitiveer, quick, accurate detection method.Micro-fluidic chip
With convenient, efficiently, the features such as high flux, but micro-fluidic chip, due to integrated function, the degree of miniaturization is not as other
Chip, therefore the direction that more small chip can be studied as micro-fluidic chip from now on.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
God any modification, equivalent substitution and improvements made etc., should be included in the scope of the protection with principle.
Claims (2)
- A kind of 1. application of micro-fluidic chip in microorganism detection.
- 2. application of the micro-fluidic chip in microorganism detection as described in the appended claim 1, it is characterised in that：Including：One micro- Fluidic chip carrys out detection bacterium DNA；Chip is prepared by dimethyl silicone polymer (PDMS), and it includes SPE (SPE) room, two independent passages and multiloop mediated isothermality amplification (LAMP) room.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711353932.4A CN107841535A (en) | 2017-12-15 | 2017-12-15 | A kind of application of micro-fluidic chip in microorganism detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711353932.4A CN107841535A (en) | 2017-12-15 | 2017-12-15 | A kind of application of micro-fluidic chip in microorganism detection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107841535A true CN107841535A (en) | 2018-03-27 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201711353932.4A Withdrawn CN107841535A (en) | 2017-12-15 | 2017-12-15 | A kind of application of micro-fluidic chip in microorganism detection |
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| CN (1) | CN107841535A (en) |
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2017
- 2017-12-15 CN CN201711353932.4A patent/CN107841535A/en not_active Withdrawn
Non-Patent Citations (1)
| Title |
|---|
| ZHE GUO ET AL.: "An integrated microfluidic chip for the detection of bacteria – A proof of concept", 《MOLECULAR AND CELLULAR PROBES》 * |
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Application publication date: 20180327 |