CN208104419U - Detect the micro-fluidic chip and detection system of human virus' nucleic acid - Google Patents
Detect the micro-fluidic chip and detection system of human virus' nucleic acid Download PDFInfo
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- CN208104419U CN208104419U CN201721919821.0U CN201721919821U CN208104419U CN 208104419 U CN208104419 U CN 208104419U CN 201721919821 U CN201721919821 U CN 201721919821U CN 208104419 U CN208104419 U CN 208104419U
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Abstract
The utility model discloses a kind of micro-fluidic chips and detection system for detecting human virus' nucleic acid.The micro-fluidic chip includes a plurality of chip bodies along chip circle distribution, the chip body includes viral nucleic acid extraction unit and PCR amplification unit, the viral nucleic acid extraction unit includes the sample charging mechanism being sequentially communicated, Solid Phase Extraction mechanism, wiper mechanism and elution mechanism, the PCR amplification unit is connected to viral nucleic acid extraction unit, the PCR amplification unit includes a plurality of PCR reaction cavities, the viral nucleic acid extraction unit is at least to extract the corresponding viral nucleic acid in sample to be tested, viral nucleic acid of the PCR amplification unit at least to extract to viral nucleic acid extraction unit expands.The micro-fluidic chip of the utility model realize viral nucleic acid automatically extract, purify and quantitative fluorescent PCR reaction, the manufacture and testing cost of chip can be reduced；A possibility that detection sensitivity is high simultaneously, can avoid missing inspection.
Description
Technical field
The utility model relates to a kind of micro-fluidic chips, in particular to a kind of to be used for hepatitis B, hepatitis C virus and people
Micro-fluidic chip that human infectious diseases' viral nucleic acid such as para-immunity defective virus quickly detects, detection method and comprising the chip
DNA rapidly extracting and detection system and device, belong to technical field of molecular biology.
Background technique
Microfluidic chip analysis is the important component in micro-total analysis system or chip lab field, final mesh
Be all reactions that pattern detection is completed on a micro-fluidic chip, pre-treatment, reaction including sample, product separation and
Detection etc..Microfluidic chip technology refers to constructing on one piece several square centimeters even smaller chip chemical or biology
Reaction, the technology can be sample preparation, mixing, reaction, separation, detection involved by the fields such as chemistry, biological, cells
The basic operations such as culture, separation, division be integrated on the chip of a very little and carry out, controlled fluid is run through into entire microchannel
Multiple technologies process, is realized controllable chain-wales by network, various response functions to realize conventional chemical, biological
It is integrated, shorten the reaction time to realize, improves detection resolution and reduce cost.
Traditional PCR reaction is carried out in conical plastic seal pipe, realizes reactant by temperature-controlling metal plate
The thermal cycle of system, heat-conducting medium are the key factors of thermal cycle speed, and the thin and thick of conical tube body and tube wall is for thermally conductive
Rate produces limitation, and micro-fluidic chip has the characteristics that surface area is big, so that caloic propagation rate is accelerated, to produce
Quick thermal cycle, the time needed for finally shortening amplification anyway, secondly micro-fluidic chip makes sample and reaction reagent
Additional amount is greatly reduced, and saves experiment spending, therefore micro-fluidic pcr chip can be widely applied to bioscience, biology
The fields such as technology, clinical medicine, disease detection and environment measuring.
Detection for hepatitis B, hepatitis C virus and human immunodeficiency virus nucleic acid in human blood sample is inspection
The whether standard compliant conventional method of blood product is tested, currently, most common method is exactly to mention to viral nucleic acid in serum
It takes, then the viral nucleic acid molecule contained in serum is detected by the method for real-time fluorescence quantitative PCR, using routine
Requirement of the method for real-time fluorescence quantitative PCR for instrument is relatively high, needs the fluorescence quantitative PCR instrument conduct of matched price valuableness
Platform is detected, and needs to extract serum sample processing, and due to there is so how cumbersome operation, this is for work
Personnel's is more demanding, and needs to carry out quality control to each environment, and detection cycle is longer.In addition general hepatitis B
Detection is mainly in hospital and Blood Center, this just has the sensitivity of detection method a relatively high requirement, and real-time fluorescence
The sensitivity of quantitative PCR has limitation, when the viral nucleic acid molecule content in serum is less, can not detect that content is extremely low
Viral nucleic acid, it may appear that the phenomenon that false negative, this just makes the detection of blood product have a possibility that missing inspection, increases the wind of missing inspection
Danger.
Utility model content
The main purpose of the utility model is to provide a kind of micro-fluidic chips for detecting human virus' nucleic acid, existing to overcome
There is the deficiency in technology.
The another object of the utility model, which also resides in, provides a kind of detection method of human virus' nucleic acid and its used of detecting
Detection system.
For realization aforementioned invention purpose, the technical solution adopted in the utility model includes：
The utility model embodiment provides a kind of micro-fluidic chip for detecting human virus' nucleic acid comprising a plurality of edges
The chip body of chip circle distribution, the chip body include viral nucleic acid extraction unit and PCR amplification unit；
The viral nucleic acid extraction unit includes the sample charging mechanism being sequentially communicated, Solid Phase Extraction mechanism, wiper mechanism and washes
Release mechanism, the PCR amplification unit are connected to viral nucleic acid extraction unit, and the PCR amplification unit includes a plurality of PCR
Reaction cavity, the viral nucleic acid extraction unit is at least to extract the corresponding viral nucleic acid in sample to be tested, the PCR
Viral nucleic acid of the amplification unit at least to extract to viral nucleic acid extraction unit expands.
In some embodiments, the wiper mechanism and elution Distribution of Institutions are set to the sample charging mechanism and solid phase extraction
Take mechanism in opposite direction clockwise or counterclockwise.
In some embodiments, the sample charging mechanism is connected to Solid Phase Extraction mechanism by first runner, the cleaning
Mechanism is connected to Solid Phase Extraction mechanism by the 5th runner, and the elution mechanism and Solid Phase Extraction mechanism are connected by second flow channel
It is logical.
Preferably, the first obstruction runner, the second are additionally provided between the 5th runner and Solid Phase Extraction mechanism
The second obstruction runner is additionally provided between road and Solid Phase Extraction mechanism.
The utility model embodiment additionally provides a kind of detection system of human virus' nucleic acid comprising detection people above-mentioned
The micro-fluidic chip of viroid nucleic acid.
Compared with prior art, include the advantages of the utility model：
1) micro-fluidic chip of detection human virus' nucleic acid provided by the utility model utilizes the effect of centrifugal force, auxiliary rainbow
Suction phenomenon and fluid resistance phenomenon, realize the feature operation of fluid, in serum viral nucleic acid (HBV, HCV, HIV1,
The viral nucleic acids such as HIV2) automatically extract and purify and quantitative fluorescent PCR reaction process be able to reality under the action of centrifuge
Existing, and waste collection cavity integrates not on chip, this can be to avoid when PCR be reacted and is heated, the reflux of waste liquid；
2) micro-fluidic chip of detection human virus' nucleic acid provided by the utility model is in operability, sensitivity, section
Save time etc. solve the problems, such as it is of the existing technology, extraction, reaction, detection is integrated on the same chip, for collection
It extracts, react, the detection device of detection one greatly reduces the manufacture and testing cost of chip without integrated large scale equipment；
The operating time is substantially reduced simultaneously, and detects a variety of viral nucleic acids on the same chip, increases the information content of detection；
3) the micro-fluidic chip flux of detection human virus' nucleic acid provided by the utility model is high, time-consuming short, can quickly,
It is efficiently completed the detection of the hepatitis Bs such as HBV, HCV, HIV, hepatitis C virus and human immunodeficiency virus nucleic acid, is obtained in time
Know that result assists further Clinics and Practices.
4) the detection method high sensitivity of human virus' nucleic acid provided by the utility model, can detecte content down to 1
The hepatitis B virus nucleic acid molecule of a copy, will test a possibility that limit is preferably minimized, avoids missing inspection.
5) detection method of human virus' nucleic acid provided by the utility model can simplify the operating process of viral diagnosis, drop
The detection error that low manual operation introduces, the detection about sample by CCD light source it is only necessary to be irradiated, and collecting signal is to virus
The yin and yang attribute of nucleic acid is detected.
Detailed description of the invention
Fig. 1 is that a kind of structure of micro-fluidic chip for detecting human virus' nucleic acid is shown in one exemplary embodiments of the utility model
It is intended to.
Description of symbols：1- cleaning solution well, 2- cleaning solution level-one cavity, 3- sample-adding cavity, 4- sample pipetting volume hole,
The obstruction runner of 5- second, 6- eluent level-one cavity, 7- eluent well, the 4th runner of 8-, 9- eluent secondary cavity,
10- second flow channel, 11- third flow channel, 12- first runner, 13- nucleic acid Solid Phase Extraction cavity, 14- nucleic acid solid phase extraction material,
15- waste liquid gas vent, the 7th runner of 16-, 19- trouble shape channel, 17,18,20,21-PCR reaction component, 22-PCR reaction cavity,
The 8th runner of 23-, 24- first hinder runner, the 5th runner of 25-, 26- cleaning solution secondary cavity, the 6th runner of 27-, 28- chip
Main body, 29- location hole.
Specific embodiment
As previously mentioned, inventor is studied for a long period of time and largely practiced in view of many defects of the existing technology, obtain
To propose the technical solution of the utility model, i.e. the utility model assists siphonage and fluid resistance using the effect of centrifugal force
Force phenomenon realizes the feature operation of fluid, extraction, the purifying and quantitative fluorescent PCR reaction for the viral nucleic acid in serum
Process is achieved under the action of centrifuge, and the detection about sample by CCD light source it is only necessary to be irradiated, collecting signal pair
The yin and yang attribute of viral nucleic acid is detected.The technical solution, its implementation process and principle etc. will be further explained as follows
Explanation.
The one aspect of the utility model embodiment provides a kind of micro-fluidic chip for detecting human virus' nucleic acid, packet
A plurality of chip bodies along chip circle distribution are included, the chip body includes viral nucleic acid extraction unit and PCR amplification list
Member；
The viral nucleic acid extraction unit includes the sample charging mechanism being sequentially communicated, Solid Phase Extraction mechanism, wiper mechanism and washes
Release mechanism, the PCR amplification unit are connected to viral nucleic acid extraction unit, and the PCR amplification unit includes a plurality of PCR
Reaction cavity, the viral nucleic acid extraction unit is at least to extract the corresponding viral nucleic acid in sample to be tested, the PCR
Viral nucleic acid of the amplification unit at least to extract to viral nucleic acid extraction unit expands.
In some embodiments, the wiper mechanism and elution Distribution of Institutions are set to the sample charging mechanism and solid phase extraction
Take mechanism in opposite direction clockwise or counterclockwise.
In some embodiments, the sample charging mechanism is connected to Solid Phase Extraction mechanism by first runner, the cleaning
Mechanism is connected to Solid Phase Extraction mechanism by the 5th runner, and the elution mechanism and Solid Phase Extraction mechanism are connected by second flow channel
It is logical.
Preferably, the first obstruction runner, the second are additionally provided between the 5th runner and Solid Phase Extraction mechanism
The second obstruction runner is additionally provided between road and Solid Phase Extraction mechanism.
Especially preferred, described first hinders runner and second to hinder runner for " S " type runner, but not limited to this.
Hindering runner by design first in the micro-fluidic chip of the utility model is " S " type runner, so that clockwise
When centrifugation, cleaning solution by nucleic acid Solid Phase Extraction cavity, and is being centrifuged eluent clockwise after sample and lysate
Shi Buhui enters nucleic acid Solid Phase Extraction cavity.Similarly, hindering runner by design second is " S " type runner, so that counterclockwise
When centrifugation, eluent enters nucleic acid Solid Phase Extraction cavity, and cleaning solution is made not enter nucleic acid Solid Phase Extraction cavity.
In some embodiments, the sample charging mechanism is used to be added the mixture of serum sample Yu serum lysate,
Including sample-adding cavity and the sample application hole being connected to the sample-adding cavity, the outlet of the sample-adding cavity and the solid phase extract
The entrance of mechanism is taken to be connected to by the first runner.
Further, the sample to be detected includes the mixture of serum sample Yu serum lysate.
Further, the sample charging mechanism is set to the surface of the chip body.
Further, location hole is provided in the chip body.
Further, plural and described chip body is equally spaced along chip circumference.
In some embodiments, the Solid Phase Extraction mechanism includes nucleic acid Solid Phase Extraction cavity and is set to the core
The sour intracorporal nucleic acid solid phase extraction material of Solid Phase Extraction chamber.
Preferably, the nucleic acid solid phase extraction material includes silica microtrabeculae.
Further, the Solid Phase Extraction mechanism is set to the underface of the sample charging mechanism.
In some embodiments, the wiper mechanism includes cleaning solution level-one cavity and cleaning solution secondary cavity, is used for
It is added and stores cleaning solution.The cleaning solution level-one cavity is connected to cleaning solution well, and the cleaning solution level-one cavity goes out
Mouth is connected to the entrance of cleaning solution secondary cavity by the 6th runner, the outlet of the cleaning solution secondary cavity and first resistance
The entrance of runner is hindered to be connected to by the 5th runner.
Further, the wiper mechanism is set to the upper left side of the chip body.
In some embodiments, the elution mechanism includes eluent level-one cavity and eluent secondary cavity, is used for
It is added and stores eluent.The eluent level-one cavity is connected to eluent well, and the eluent level-one cavity goes out
Mouth is connected to the entrance of eluent secondary cavity by the 4th runner, the outlet of the eluent secondary cavity and second resistance
The entrance of runner is hindered to be connected to by the second flow channel, the outlet of the second obstruction runner and entering for the Solid Phase Extraction mechanism
Mouth passes through third flow channel and is connected to.
Further, the elution mechanism is set to the upper right side of the chip body.
In some embodiments, the PCR amplification unit includes trouble shape channel and mutual with the trouble shape channel respectively
A plurality of PCR reaction cavities of connection, preferably include four parallel PCR reaction cavities, in each reaction cavity, are added different
Primer and probe, be used for fluorescent quantitative PCR or signal detection.
Preferably, the outlet of the entrance and the viral nucleic acid extraction unit in the trouble shape channel is connected by the 8th runner
Logical, each outlet in the trouble shape channel is connected to the entrance of the PCR reaction cavity by the 7th runner.
Further, the PCR reaction cavity is connected to eluent cavity, is closed cavity, and inside is placed with PCR reaction
Or reaction raw materials, primer and probe needed for detection.
Preferably, it is provided with waste liquid gas vent on the 8th runner, further, the waste liquid gas vent and waste liquid are received
Collecting mechanism connection.Further, it is connected to 2mL centrifuge tube below waste liquid gas vent, connected with the waste liquid vent hole seal of chip
It connects, is connected with viral nucleic acid extraction unit, for being stored in the waste liquid generated in nucleic acid extraction and purification process, while can also be with
For exhausting remaining gas in runner.
Preferably, it is provided in the PCR reaction cavity at least to fluorescent PCR amplification or primer and the spy of signal detection
Needle.
By above-mentioned technical proposal, the micro-fluidic chip of detection human virus' nucleic acid provided by the utility model utilizes centrifugation
Effect, auxiliary siphonage and the fluid resistance phenomenon of power, realize the feature operation of fluid, for the viral nucleic acid in serum
(HBV, HCV, HIV1, HIV2 these fourth types viral nucleic acid) automatically extract and purify and quantitative fluorescent PCR reaction process from
It is achieved under the action of scheming, and waste collection cavity integrates not on chip, this can be heated to avoid reacting in PCR
When, the reflux of waste liquid；Meanwhile the utility model will be extracted, be reacted, detection is integrated on the same chip, be extracted for collection, instead
Answer, detect the detection device of one to greatly reduce the manufacture and testing cost of chip without integrated large scale equipment；It is big simultaneously
The operating time is shortened greatly, and detects a variety of viral nucleic acids on the same chip, increases the information content of detection.
The utility model embodiment another aspect provides a kind of detection systems of human virus' nucleic acid comprising it is preceding
The micro-fluidic chip for the detection human virus' nucleic acid stated.
The human virus' nucleic acid implemented based on the micro-fluidic chip above-mentioned for detecting human virus' nucleic acid or detection system
Detection method include：
Sample to be tested is added into the sample charging mechanism of viral nucleic acid extraction unit, cleaning solution is added into wiper mechanism,
Eluent is added into elution mechanism；
Under the influence of centrifugal force by chip body according to clockwise or counterclockwise, so that sample to be tested and cleaning
Liquid enters the Solid Phase Extraction mechanism and starts the cleaning processing, and eluent is hindered to enter the Solid Phase Extraction mechanism；
Under the influence of centrifugal force by chip body according to counterclockwise or rotate clockwise so that eluent enter it is described solid
Phase extraction mechanism carries out elution processing, and cleaning solution is hindered to enter the Solid Phase Extraction mechanism；
Later, under the influence of centrifugal force by chip body according to counterclockwise or rotate clockwise, make elution treated
Sample to be tested enters PCR amplification unit, carries out fluorescent PCR amplification or signal detection.
Among some more preferably exemplary embodiments, the detection method can specifically comprise the following steps：
1. the mixture of human serum sample and lysate is added into sample-adding cavity from sample pipetting volume hole, from cleaning solution plus
Cleaning buffer solution (wash buffer) 1 and cleaning buffer solution 2 are added into cleaning solution level-one cavity for sample hole, are loaded from eluent
Eluent is added into eluent level-one cavity in hole；
2. rotate clockwise centrifugation 3min with the rate of 2000rpm, the mixture of human serum sample and lysate will be through
Enter nucleic acid Solid Phase Extraction cavity after first runner, cleaning solution enters by the 6th runner, cleaning solution secondary cavity, the 5th runner
Nucleic acid Solid Phase Extraction cavity；
3. the collecting pipe of waste liquid gas vent of dismantling connection, blend compounds pad seal waste liquid gas vent；
4. with the rate of 50rpm rotating centrifugal counterclockwise 30 seconds, so that eluent enters nucleic acid Solid Phase Extraction cavity；
5. with 2500rpm high speed rotating centrifugal 1 minute counterclockwise, with powerful centrifugal force make the nucleic acid solution after elution into
Enter PCR reaction cavity, and in conjunction with PCR Master Mix, primer, probe.
6. chip is put into flat heating device and carries out PCR amplification；
7. the chip that amplification is completed, which is placed under CCD light source, carries out fluorescence detection.
The detection method high sensitivity of the utility model can detecte the hepatitis B virus nucleic acid that content is copied down to 1
Molecule will test a possibility that limit is preferably minimized, avoids missing inspection；Also, the utility model can simplify the operation of viral diagnosis
Process reduces the detection error that manual operation introduces, and the detection about sample collects letter it is only necessary to irradiate by CCD light source
Number the yin and yang attribute of viral nucleic acid is detected.
Clear, complete description is carried out to the technical solution of the utility model below in conjunction with attached drawing and typical case.
Please refer to the micro-fluidic chip that Fig. 1 show one of the present embodiment detection human virus' nucleic acid comprising core
Piece main body 28, the chip body 28 include：Viral nucleic acid extraction unit comprising interconnected sample charging mechanism, solid phase extraction
Mechanism, wiper mechanism and elution mechanism are taken, at least to make sample to be tested in conjunction with solid phase carrier under the influence of centrifugal force,
And complete cleaning, elution processing；And the PCR amplification unit being interconnected with the viral nucleic acid extraction unit comprising multiple
Several PCR reaction cavities, each PCR reaction cavity are connected with the outlet of the viral nucleic acid extraction unit, at least use
With fluorescent PCR amplification or signal detection.
Wherein, the sample charging mechanism is set to the surface of the chip body 28 comprising sample-adding cavity 3 and with institute
The sample application hole 4 that sample-adding cavity 3 is connected to is stated, the outlet of the sample-adding cavity 3 and the entrance of the Solid Phase Extraction mechanism pass through
The first runner 12 is connected to.Preferably, the sample to be detected includes the mixture of serum sample Yu serum lysate.Its
In, the location hole 29 for facilitating positioning is additionally provided in the chip body 28.
The Solid Phase Extraction mechanism is set to the underface of the sample charging mechanism comprising nucleic acid Solid Phase Extraction cavity 13 with
And it is set to the nucleic acid solid phase extraction material 14 in nucleic acid Solid Phase Extraction cavity 13, such as silica microtrabeculae.
The wiper mechanism is set to the upper left side of chip body 28 comprising cleaning solution level-one cavity 2 and cleaning solution two
Grade cavity 26, the cleaning solution level-one cavity 2 be connected to cleaning solution well 1, the outlet of the cleaning solution level-one cavity 2 with clearly
The entrance of washing lotion secondary cavity 26 is connected to by the 6th runner 27, and the outlet of the cleaning solution secondary cavity 26 hinders to flow with first
The entrance in road 24 is connected to by the 5th runner 25.
The elution mechanism is set to the upper right side of chip body 28 comprising eluent level-one cavity 6 and eluent two
Grade cavity 9, the eluent level-one cavity 6 is connected to eluent well 7, the outlet of the eluent level-one cavity 6 with wash
The entrance of de- liquid secondary cavity 9 is connected to by the 4th runner 8, and the outlet of the eluent secondary cavity 9 hinders runner 5 with second
Entrance be connected to by second flow channel 10, described second hinders the entrance of the outlet of runner 5 and nucleic acid Solid Phase Extraction cavity 13 logical
Cross the connection of third flow channel 11.
Hindering runner 24 by design first in the micro-fluidic chip of the utility model is " S " type runner, so that in up time
When needle is centrifuged, cleaning solution by nucleic acid Solid Phase Extraction cavity 13, and makes eluent clockwise after sample and lysate
Nucleic acid Solid Phase Extraction cavity 13 will not be entered when centrifugation.Similarly, hindering runner 5 by design second is " S " type runner, so that
Counterclockwise when centrifugation, eluent enters nucleic acid Solid Phase Extraction cavity 13, and cleaning solution is made not enter nucleic acid Solid Phase Extraction chamber
Body 13.
Wherein, the PCR amplification unit include trouble shape channel 19 (in the present embodiment for four trouble channels) and respectively with branch off shape
In each reaction cavity, different PCR reaction components is added in four interconnected parallel PCR reaction cavities 22 of channel 19
17,18,20,21, including reaction raw materials, primer and probe, it is used for fluorescent quantitative PCR or signal detection.The trouble shape is logical
The entrance in road 19 is connected to the outlet of nucleic acid Solid Phase Extraction cavity 13 by the 8th runner 23, each outlet in the trouble shape channel 19
It is connected to the entrance of the PCR reaction cavity 22 by the 7th runner 16.
Wherein, it is provided with waste liquid gas vent 15 on the 8th runner 23,2mL centrifugation is connected to below waste liquid gas vent
Pipe, connect with the waste liquid vent hole seal of chip body, is connected with viral nucleic acid extraction unit, for being stored in nucleic acid extraction
With the waste liquid generated in purification process, while it can also be used to exhaust remaining gas in runner.
Wherein, chip body is made by PC material in the present embodiment, radius 40mm, with a thickness of 2mm, wherein cleaning
The volume of liquid level-one cavity 2 and cleaning solution secondary cavity 26 is 600 μ l, and the volume of sample-adding cavity 3 is 500 μ l, eluent level-one
The volume of cavity 6 and eluent secondary cavity 9 is 40 μ l, and waste liquid gas vent 15 connects the sewer pipe that volume is 2000 μ l.
The specific step that human virus' nucleic acid in sample to be tested is detected using the micro-fluidic chip of the present embodiment
Suddenly include：
Step 1：The micro-fluidic chip that will test human virus' nucleic acid takes out from -20 DEG C of refrigerators, reacts component to PCR
17, after 18,20,21 thawings, chip body 28 is flat on the adapter of centrifuge, it is solid by location hole 29 and adapter
It is fixed.It should be noted and keep central symmetry in chip main body 28.
Step 2：The 400 μ L of mixture of sample and lysate is added through sample pipetting volume hole 4, cleaned liquid well 1 is added
30 μ L of eluent is added through eluent well 7 in 300 μ Lwash buffer 1 and 200 μ Lwash buffer 2.
Step 3：Centrifugation 3min is rotated clockwise with the rate of 2000rpm, sample will be through first-class with cracking liquid mixture
Enter nucleic acid Solid Phase Extraction cavity 13 behind road 12, cleaning solution passes through the 6th runner 27, cleaning solution secondary cavity 26, the 5th runner 25
And " S " type first hinders runner 24 to enter nucleic acid Solid Phase Extraction cavity 13；Waste liquid after cleaning is entered by drain gas vent 15
Waste collection pipe.
Step 4：The collecting pipe of waste liquid gas vent of dismantling connection, blend compounds pad seal waste liquid gas vent.
Step 5：With the rate of 50rpm rotating centrifugal counterclockwise 30 seconds, so that eluent passes through the 4th runner 8, eluent
Secondary cavity 9, second flow channel 10 and " S " type second hinder runner 5 to enter nucleic acid Solid Phase Extraction cavity 13.
Step 6：With 2500rpm high speed rotating centrifugal 1 minute counterclockwise, keep the nucleic acid of elution molten with powerful centrifugal force
Liquid by the 8th runner 23, trouble shape channel 19, the 7th runner 16, into PCR reaction cavity 22, and with PCR MasterMix, draw
Object, probe mixing.
Step 7：Chip body is put into flat heating device and carries out PCR amplification.
Step 8：The chip that amplification is completed, which is placed under CCD light source, carries out fluorescence detection.
By above-mentioned technical proposal, the micro-fluidic chip of the utility model realize viral nucleic acid automatically extract and purify with
And quantitative fluorescent PCR reaction, without integrated large scale equipment, the manufacture and testing cost of chip can be substantially reduced；Spirit is detected simultaneously
A possibility that sensitivity is high, can avoid missing inspection.
The technology contents and technical characteristic of the utility model have revealed that as above, however those skilled in the art still may be used
Can the announcement based on the utility model and make various replacements and modification without departing substantially from the spirit of the present invention, therefore, this is practical new
Type protection scope should be not limited to the revealed content of embodiment, and including the various replacements without departing substantially from the utility model and should repair
Decorations, and covered by present patent application claim.
Claims (10)
1. a kind of micro-fluidic chip for detecting human virus' nucleic acid, it is characterised in that including a plurality of cores along chip circle distribution
Piece main body, the chip body include viral nucleic acid extraction unit and PCR amplification unit；
The viral nucleic acid extraction unit includes the sample charging mechanism being sequentially communicated, Solid Phase Extraction mechanism, wiper mechanism and washing and dehydrating integrated machine
Structure, the PCR amplification unit are connected to viral nucleic acid extraction unit, and the PCR amplification unit includes a plurality of PCR reactions
Cavity, the viral nucleic acid extraction unit is at least to extract the corresponding viral nucleic acid in sample to be tested, the PCR amplification
Viral nucleic acid of the unit at least to extract to viral nucleic acid extraction unit expands.
2. micro-fluidic chip according to claim 1, it is characterised in that：The wiper mechanism and elution Distribution of Institutions setting
In the sample charging mechanism and Solid Phase Extraction mechanism in opposite direction clockwise or counterclockwise.
3. micro-fluidic chip according to claim 1 or 2, it is characterised in that：The sample charging mechanism and Solid Phase Extraction mechanism
It is connected to by first runner, the wiper mechanism is connected to Solid Phase Extraction mechanism by the 5th runner, the elution mechanism and solid
Phase extraction mechanism is connected to by second flow channel.
4. micro-fluidic chip according to claim 3, it is characterised in that：Between 5th runner and Solid Phase Extraction mechanism
It is additionally provided with the first obstruction runner, the second obstruction runner is additionally provided between the second flow channel and Solid Phase Extraction mechanism, it is described
First hinders runner and second to hinder runner for " S " type runner.
5. micro-fluidic chip according to claim 3, it is characterised in that：The sample charging mechanism include sample-adding cavity and with
The sample application hole of the sample-adding cavity connection, the outlet of the sample-adding cavity and the entrance of the Solid Phase Extraction mechanism pass through institute
First runner connection is stated, the sample charging mechanism is set to the surface of the chip body, and it is fixed to be provided in the chip body
Position hole, a plurality of chip bodies are equally spaced along chip circumference.
6. micro-fluidic chip according to claim 3, it is characterised in that：The Solid Phase Extraction mechanism includes nucleic acid solid phase extraction
It takes cavity and is set to the intracorporal nucleic acid solid phase extraction material of the nucleic acid Solid Phase Extraction chamber, the Solid Phase Extraction mechanism setting
In the underface of the sample charging mechanism.
7. micro-fluidic chip according to claim 4, it is characterised in that：The wiper mechanism includes cleaning solution level-one cavity
With cleaning solution secondary cavity, the cleaning solution level-one cavity is connected to cleaning solution well, and the cleaning solution level-one cavity goes out
Mouth is connected to the entrance of cleaning solution secondary cavity by the 6th runner, the outlet of the cleaning solution secondary cavity and first resistance
The entrance of runner is hindered to be connected to by the 5th runner, the wiper mechanism is set to the upper left side of the chip body.
8. micro-fluidic chip according to claim 4, it is characterised in that：The elution mechanism includes eluent level-one cavity
With eluent secondary cavity, the eluent level-one cavity is connected to eluent well, and the eluent level-one cavity goes out
Mouth is connected to the entrance of eluent secondary cavity by the 4th runner, the outlet of the eluent secondary cavity and second resistance
The entrance of runner is hindered to be connected to by the second flow channel, the outlet of the second obstruction runner and entering for the Solid Phase Extraction mechanism
Mouth passes through third flow channel and is connected to, and the elution mechanism is set to the upper right side of the chip body.
9. according to claim 1,2, micro-fluidic chip described in any one of 4-8, it is characterised in that：The PCR amplification unit
The a plurality of PCR reaction cavities interconnected with the trouble shape channel including trouble shape channel and respectively, the trouble shape channel enters
Mouth is connected to the outlet of the viral nucleic acid extraction unit by the 8th runner, each outlet in the trouble shape channel and the PCR
The entrance of reaction cavity is connected to by the 7th runner, is provided with waste liquid gas vent, the waste liquid gas vent on the 8th runner
Mechanism is connect with waste collection；The primer at least to fluorescent PCR amplification or signal detection is provided in the PCR reaction cavity
And probe.
10. a kind of detection system of human virus' nucleic acid, it is characterised in that including detection of any of claims 1-9
The micro-fluidic chip of human virus' nucleic acid.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107893026A (en) * | 2017-12-29 | 2018-04-10 | 苏州绘真医学检验所有限公司 | Detect micro-fluidic chip, detection method and the detecting system of human virus' nucleic acid |
| WO2020192169A1 (en) * | 2019-03-27 | 2020-10-01 | 广州万孚生物技术股份有限公司 | In vitro detection device and sample loading mechanism thereof |
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2017
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107893026A (en) * | 2017-12-29 | 2018-04-10 | 苏州绘真医学检验所有限公司 | Detect micro-fluidic chip, detection method and the detecting system of human virus' nucleic acid |
| WO2020192169A1 (en) * | 2019-03-27 | 2020-10-01 | 广州万孚生物技术股份有限公司 | In vitro detection device and sample loading mechanism thereof |
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