CN208200972U - A kind of sample automatically processes and nucleic acid amplifier - Google Patents
A kind of sample automatically processes and nucleic acid amplifier Download PDFInfo
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- CN208200972U CN208200972U CN201820196093.3U CN201820196093U CN208200972U CN 208200972 U CN208200972 U CN 208200972U CN 201820196093 U CN201820196093 U CN 201820196093U CN 208200972 U CN208200972 U CN 208200972U
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Abstract
Automatically processed the utility model discloses a kind of sample and nucleic acid amplifier, mainly include it is multiple for nucleic acid amplification agents save with react circular chamber, multiple cylindrical chambers for being used for sample processing reagent preservation and reacting；Whole structure in the form of annular discs, the disc-shaped structure divides inner ring and outer ring two parts, inner ring can carry out the horizontal rotation relative to outer ring centered on axis, it is multiple for nucleic acid amplification agents preservation disc-shaped structure outer ring is evenly distributed on circular chamber react, it is multiple be used for sample processing reagent preservation in the cylindrical chamber reacted.Traditional manual extraction purification of nucleic acid mode is organically integrated into full automatic enclosed workshop process by the utility model, cut down cumbersome manual steps, potential false positive pollution risk is efficiently avoided, and operating process is quick, is detected convenient for high-volume sample number.
Description
Technical field
The utility model relates to the fields of the nucleic acid extraction of molecular biology and amplified reaction, and in particular to it is a kind of automatically from
The device for extracting nucleic acid in sample and being expanded.
Background technique
Nucleic acid extraction and amplification are the important method steps of biomedical research and clinical molecular diagnosis work.In clinic point
Son is examined in practical application, is frequently necessary to handle more sample simultaneously, and nucleic acid extraction isolation technics conventional at present includes
Precipitating, a series of processes such as centrifugation, step is complicated, takes time and effort, low efficiency, it is difficult to which satisfaction mentions high-throughput clinical sample
Take the work requirements of purifying.Meanwhile carrying out polymerase chain reaction (the Polymerase Chain for amplification of nucleic acid
Reaction, abbreviation PCR) and the operation of constant-temperature amplification formula reaction experiment in, need various reacted constituents through manual mode such as
Enzyme, primer and probe etc. move into nucleic acid amplification reaction pipe from different memotrons, cumbersome, are easy error, are unfavorable for big
The nucleic acid amplification reaction of scale high throughput, limits the development of molecular diagnostic techniques.
Therefore, to improve working efficiency, nucleic acid can be extracted automatically and be expanded from sample by needing to design and develop one kind
The consumable material device of increasing can automatically complete whole treatment processes of nucleic acid extraction amplification in matched instrument and equipment.
Utility model content
The purpose of the utility model is to overcome the shortcomings of the prior art, and provide a kind of sample and automatically process and core
Sour amplification device.
The purpose of this utility model is completed by following technical solution: this sample automatically processes and nucleic acid amplification
Device and application method, mainly include it is multiple for nucleic acid amplification agents save with react circular chamber, it is multiple be used for sample
Cylindrical chamber reagent treatment preservation and reacted；Whole structure in the form of annular discs, the disc-shaped structure divide inner ring and outer ring
Two parts, inner ring can carry out the horizontal rotation relative to outer ring centered on axis, multiple to save for nucleic acid amplification agents
It is evenly distributed on the circular chamber reacted on disc-shaped structure outer ring, saves and react for sample processing reagent multiple
In cylindrical chamber, there is a central reaction chamber to be located at disc-shaped structure inner ring center, remaining cylindrical chamber is uniformly distributed
In disc-shaped structure outer ring, with multiple circular chamber's cross-distributions for nucleic acid amplification agents preservation and reacting.Material is poly-
Allyl resin (PP) plastics or polycarbonate (PC) resin and plastic or polyethylene (PE) resin and plastic.
The cylindrical center reaction chamber, which includes, to control the rubber plunger and adsorbable nucleic acid that move up and down by external force
Nanometer magnetic bead, the cylindrical chamber and use for saving and reacting for sample processing reagent of central reaction cavity bottom and outer ring
It is saved in nucleic acid amplification agents and passes through with the circular chamber reacted that microchannel is radial to be connected respectively, central reaction chamber
Bottom is inner ring side wall, contains 1 microchannel aperture, can make central reaction chamber and other chambers by the horizontal rotation of inner ring
The multiple microchannels being connected are opened one by one.
The cylindrical chamber for being distributed in outer ring sequentially includes with lower chambers in the direction of the clock:
1) lytic reagent chamber is mounted with lytic reagent, and the lytic reagent includes at least one: guanidine salt, chaotropic
Salt, red blood cell lysis reagent, detergent, chelating agent, sodium hydroxide, dnase inhibitor, RNase inhibitor, anti-coagulants, blood coagulation
Agent, protease, surfactant.
2) washing reagent chamber (2 pipe), is mounted with washing reagent, the washing reagent includes at least one: second
Alcohol, sodium chloride, Tris hydrochloric acid.
3) elution reagent chamber, is mounted with elution reagent, and the elution reagent includes at least Tris hydrochloric acid.
The circular chamber's top sealer for being used for nucleic acid amplification agents and saving and reacting, leaks outside to prevent reagent, in chamber
Equipped with nucleic acid constant-temperature amplification or PCR amplification reagent, the reagent includes at least one: Bst polymerase, Taq polymerase, inverse
Transcriptase, four kinds of nucleotide monomers (dNTPs), primers, probe.
The sample includes one of at least following substance: cell, spore, microorganism, organism biopsy, excrement, life
Thing liquid, soil and ambient water.
The cell is the intracorporal cell of biology and laboratory cultures cell, the microorganism be virus, bacterium, mould,
Helminth, the biological fluids are allantoic fluid, bile, cholic acid, cholate, BILE PIGMENTS, blood, blood plasma, serum, cerebrospinal fluid, chorion
Liquid, colostrum, digestive juice, exudate, omission timber, hemolymph, lochia, lymph, chyle, milk, hydrothorax, saliva, suet, sperm,
Phlegm, synovial fluid, tear, urine or vaginal secretion.
The exudate is amniotic fluid, ascites or sweat, and the omission timber is pericardial fluid or peritoneal fluid, and the digestive juice is stomach
Liquid, intestinal juice or pancreatic juice.
The utility model provides a kind of processing method automatically extracted with amplified sample nucleic acid amplification, and this method includes such as
Lower step:
1) opens the rubber plunger of lytic reagent chamber, 100-500 microlitres of liquid sample is added, to liquid sample stream
Rubber plunger is closed after entering, and makes lysate and the liquid sample hybrid reaction in chamber, liquid sample cracking release nucleic acid；
2) rotates horizontally inner ring, and 1 microchannel aperture of central reaction cavity bottom is made to correspond to connection lytic reagent
The microchannel of chamber；
3) moves down the rubber plunger of lytic reagent chamber by external force, while moving up central reaction chamber
The rubber plunger of room makes the mixing liquid of lytic reagent chamber enter central reaction chamber by microchannel, in mixing liquid
Nucleic acid in conjunction with the magnetic bead of central reaction chamber；
4) moves up the rubber plunger of lytic reagent chamber by external force, while moving down central reaction chamber
The rubber plunger of room makes mixing liquid return to lytic reagent chamber by microchannel, and the magnetic bead of nucleic acid is combined to pass through center
Magnet attraction outside reaction chamber stays in central reaction cavity bottom；
5) rotates horizontally inner ring, and 1 microchannel aperture of central reaction cavity bottom is made to correspond to one washing of connection
The microchannel of reagent chamber；
6) moves down the rubber plunger of washing reagent chamber by external force, while moving up central reaction chamber
The washing reagent of the rubber plunger of room, washing reagent chamber enters central reaction chamber by microchannel, makes to have combined nucleic acid
Magnetic bead immerse cleaning solution, washing removal impurity；
7) moves up the rubber plunger of washing reagent chamber by external force, while moving down central reaction chamber
The rubber plunger of room, the liquid after making washing returns to washing reagent chamber by microchannel, and the magnetic bead of nucleic acid is combined to pass through
Magnet attraction outside central reaction chamber stays in central reaction cavity bottom；
8) rotates horizontally inner ring, and 1 microchannel aperture of the central reaction cavity bottom is made to correspond to connection the
The microchannel of two washing reagent chambers；
6) and 7) 9) repeats the above steps, and carries out second and washs；
10) rotates horizontally inner ring, and 1 microchannel aperture of the central reaction cavity bottom is made to correspond to connection
The microchannel of elution reagent chamber；
11) moves down the rubber plunger of elution reagent chamber by external force, while moving up central reaction chamber
The elution reagent of the rubber plunger of room, elution reagent chamber enters central reaction chamber by microchannel, makes to have combined nucleic acid
Magnetic bead immerse eluent, sample nucleic acid be detached from magnetic bead, be dissolved in eluent；
12) rotates horizontally inner ring, and 1 microchannel aperture of central reaction cavity bottom is made to correspond to connection first
Nucleic acid amplification agents save the microchannel with reaction chamber；
13) moves down the rubber plunger of central reaction chamber by external force, partially containing the elution reagent of nucleic acid
Enter first nucleic acid amplification agents preservation and reaction chamber by microchannel, mixes nucleic acid with nucleic acid amplification agents；
14) rotates horizontally inner ring, and 1 microchannel aperture of central reaction cavity bottom is made to correspond to connection second
Nucleic acid amplification agents save the microchannel with reaction chamber；
15) 13). repeats the above steps, mixes nucleic acid and the nucleic acid amplification agents of second chamber；
14) and 15) 16) repeats the above steps, and makes that nucleic acid and third, the 4th nucleic acid amplification agents save and reaction chamber
The nucleic acid amplification agents of room mix；
17) carries out fluorescent PCR nucleic acid amplification reaction according to the amplification program of setting or constant temperature nucleic acid amplification reacts, and
It further include that DNA is synthesized by reverse transcription.
The utility model has the following beneficial effects: the utility model is organically whole by traditional manual extraction purification of nucleic acid mode
It is combined into full automatic enclosed workshop process, cuts down cumbersome manual steps, it is dirty to efficiently avoid potential false positive
Risk is contaminated, and operating process is quick, is detected convenient for high-volume sample number.
Detailed description of the invention
Fig. 1 is the constructional appearance figure of the utility model.
Fig. 2 is the outer ring structure schematic diagram of the utility model.
Fig. 3 is the structure top view of the utility model.
Description of symbols: outer ring 1, central reaction chamber 2, PCR reaction chamber 3, cleaning solution A4, eluent 5, cleaning solution
B6, eddy channel platen 7, adding mouth 8.
Specific embodiment
Detailed introduction is done to the utility model below in conjunction with attached drawing:
As shown in the picture, this sample automatically process with nucleic acid amplifier and application method, it is main to include 4 and be used for core
Sour circular chamber amplifing reagent preservation and reacted, 5 cylindrical chambers for saving and reacting for sample processing reagent are described
It is disc-shaped structure that sample, which is automatically processed with nucleic acid amplifier appearance, and material is acrylic resin (PP) plastics or polycarbonate
(PC) resin and plastic or polyethylene (PE) resin and plastic.
The disc-shaped structure divides inner ring and outer ring 1 two parts, and inner ring can be carried out centered on axis relative to outer ring 1
It rotates horizontally, 4 are evenly distributed on disc-shaped structure outer ring 1 with the circular chamber reacted for nucleic acid amplification agents preservation, 5
It is a to save for sample processing reagent with the cylindrical chamber reacted, there is a central reaction chamber 2 to be located at disc-shaped structure
Inner ring center, remaining 4 cylindrical chamber are evenly distributed on disc-shaped structure outer ring 1, save with 4 for nucleic acid amplification agents
With the circular chamber's cross-distribution reacted.
The cylindrical center reaction chamber 2, which includes, to control the rubber plunger and adsorbable nucleic acid that move up and down by external force
Nanometer magnetic bead, 4 cylindric chambers for saving and reacting for sample processing reagent of 2 bottom of central reaction chamber and outer ring 1
It is radial connected that room and 4 for nucleic acid amplification agents preservation pass through 8 microchannels with the circular chamber reacted respectively
It connects, 2 bottom of central reaction chamber is inner ring side wall, contains 1 microchannel aperture, center can be kept anti-by the horizontal rotation of inner ring
8 microchannels for answering chamber 2 to be connected with other chambers are opened one by one.
The cylindrical center reaction chamber 2 includes respectively to control the rubber plunger moved up and down by external force.
Described 4 cylindrical chambers for being distributed in outer ring 1 sequentially include with lower chambers in the direction of the clock:
1) lytic reagent chamber is mounted with lytic reagent, and the lytic reagent includes at least one: guanidine salt, chaotropic
Salt, red blood cell lysis reagent, detergent, chelating agent, sodium hydroxide, dnase inhibitor, RNase inhibitor, anti-coagulants, blood coagulation
Agent, protease, surfactant.
2) washing reagent chamber (2 pipe), is mounted with washing reagent, the washing reagent includes at least one: second
Alcohol, sodium chloride, Tris hydrochloric acid.
3) elution reagent chamber, is mounted with elution reagent, and the elution reagent includes at least Tris hydrochloric acid.
The circular chamber's top sealer for being used for nucleic acid amplification agents and saving and reacting, leaks outside to prevent reagent, in chamber
Equipped with nucleic acid constant-temperature amplification or PCR amplification reagent, the reagent includes at least one: Bst polymerase, Taq polymerase, inverse
Transcriptase, four kinds of nucleotide monomers (dNTPs), primers, probe.
The sample includes one of at least following substance: cell, spore, microorganism, organism biopsy, excrement, life
Thing liquid, soil and ambient water.
The cell is the intracorporal cell of biology and laboratory cultures cell, the microorganism be virus, bacterium, mould,
Helminth, the biological fluids are allantoic fluid, bile, cholic acid, cholate, BILE PIGMENTS, blood, blood plasma, serum, cerebrospinal fluid, chorion
Liquid, colostrum, digestive juice, exudate, omission timber, hemolymph, lochia, lymph, chyle, milk, hydrothorax, saliva, suet, sperm,
Phlegm, synovial fluid, tear, urine or vaginal secretion.
The exudate is amniotic fluid, ascites or sweat, and the omission timber is pericardial fluid or peritoneal fluid, and the digestive juice is stomach
Liquid, intestinal juice or pancreatic juice.
Embodiment 1:
The rubber plunger of lytic reagent chamber is opened, 100-500 microlitres of liquid sample is added, after liquid sample inflow
Rubber plunger is closed, lysate and the liquid sample hybrid reaction in chamber, liquid sample cracking release nucleic acid are made.
Inner ring is rotated horizontally, 1 microchannel aperture of 2 bottom of central reaction chamber is made to correspond to connection lytic reagent chamber
The microchannel of room.Lysate is aligned with the cylinder slot of liquid sample.
External force pushes down on the rubber plunger of lytic reagent chamber, while moving up the rubber column of central reaction chamber 2
Plug makes the mixing liquid of lytic reagent chamber enter central reaction chamber 2 by microchannel, and the intracorporal nucleic acid of mixed liquor is in
The magnetic bead of heart reaction chamber 2 combines.
External force moves up the rubber plunger of lytic reagent chamber, while pushing down on the rubber column of central reaction chamber 2
Plug makes mixing liquid return to lytic reagent chamber by microchannel, and the magnetic bead of nucleic acid is combined to pass through outside central reaction chamber 2
Magnet attraction stay in central reaction cavity bottom.
Inner ring is rotated horizontally, 1 microchannel aperture of 2 bottom of central reaction chamber is made to correspond to first washing of connection
The microchannel of reagent chamber.
External force pushes down on the rubber plunger of first washing reagent chamber, while moving up central reaction chamber 2
The washing reagent of rubber plunger, first washing reagent chamber enters central reaction chamber 2 by microchannel, makes syncaryon
The magnetic bead of acid immerses cleaning solution, washing removal impurity.
External force moves up the rubber plunger of first washing reagent chamber, while pushing down on central reaction chamber 2
Rubber plunger, the liquid after making washing returns to first washing reagent chamber by microchannel, and combines the magnetic bead of nucleic acid logical
It crosses magnet attraction and stays in 2 bottom of central reaction chamber.
Inner ring is rotated horizontally, 1 microchannel aperture of 2 bottom of central reaction chamber is made to correspond to second washing of connection
The microchannel of reagent chamber.
External force pushes down on the rubber plunger of second washing reagent chamber, while moving up central reaction chamber 2
The washing reagent of rubber plunger, second washing reagent chamber enters central reaction chamber 2 by microchannel, makes syncaryon
The magnetic bead of acid immerses cleaning solution, washing removal impurity.
External force moves up the rubber plunger of second washing reagent chamber, while pushing down on central reaction chamber 2
Rubber plunger, the liquid after making washing returns to second washing reagent chamber by microchannel, and combines the magnetic bead of nucleic acid logical
It crosses magnet attraction and stays in 2 bottom of central reaction chamber.
Inner ring is rotated horizontally, 1 microchannel aperture of 2 bottom of central reaction chamber is made to correspond to connection elution reagent chamber
The microchannel of room.
External force pushes down on the rubber plunger of elution reagent chamber, while moving up the rubber of central reaction chamber 2
The elution reagent of rubber column gel column plug, elution reagent chamber enters central reaction chamber 2 by microchannel, makes the magnetic bead for having combined nucleic acid
Eluent is immersed, sample nucleic acid is detached from magnetic bead, is dissolved in eluent.
Inner ring is rotated horizontally, 1 microchannel aperture of 2 bottom of central reaction chamber is made to correspond to first nucleic acid of connection
Amplifing reagent saves the microchannel with reaction chamber.
External force pushes down on the rubber plunger of central reaction chamber, and partially the elution reagent containing nucleic acid passes through microchannel
Into first nucleic acid amplification agents preservation and reaction chamber, mix nucleic acid with nucleic acid amplification agents therein.
Inner ring is rotated horizontally, 1 microchannel aperture of 2 bottom of central reaction chamber is made to correspond to second nucleic acid of connection
Amplifing reagent saves the microchannel with reaction chamber.
External force pushes down on the rubber plunger of central reaction chamber 2, and partially the elution reagent containing nucleic acid is logical by miniflow
Road enters second nucleic acid amplification agents preservation and reaction chamber, mixes nucleic acid with nucleic acid amplification agents therein.
Inner ring is rotated horizontally, 1 microchannel aperture of 2 bottom of central reaction chamber is made to correspond to connection third nucleic acid
Amplifing reagent saves the microchannel with reaction chamber.
External force pushes down on the rubber plunger of central reaction chamber, and partially the elution reagent containing nucleic acid passes through microchannel
Into the preservation of third nucleic acid amplification agents and reaction chamber, mix nucleic acid with nucleic acid amplification agents therein.
Inner ring is rotated horizontally, 1 microchannel aperture of 2 bottom of central reaction chamber is made to correspond to the 4th nucleic acid of connection
Amplifing reagent saves the microchannel with reaction chamber.
External force pushes down on the rubber plunger of central reaction chamber, and partially the elution reagent containing nucleic acid passes through microchannel
Into the 4th nucleic acid amplification agents preservation and reaction chamber, mix nucleic acid with nucleic acid amplification agents therein.
Nucleic acid fluorescent PCR amplification or fluorescence constant-temperature amplification are carried out according to the amplification program of setting.
It is understood that it will be understood by those skilled in the art that the technical solution of the utility model and utility model structure
Think of is subject to equivalent substitution or change all and should belong to the protection scope of the utility model the attached claims.
Claims (2)
1. a kind of sample automatically processes and nucleic acid amplifier, it is characterised in that: mainly include multiple for nucleic acid amplification agents
Save with circular chamber react, multiple be used for sample processing reagent preservation and the cylindrical chamber that reacts；It is whole in the form of annular discs
Structure, the disc-shaped structure divide inner ring and outer ring (1) two parts, and inner ring can be carried out centered on axis relative to outer ring
(1) horizontal rotation, it is multiple to be evenly distributed on outside disc-shaped structure for nucleic acid amplification agents preservation with the circular chamber reacted
Enclose (1) on, it is multiple for sample processing reagent save and in the cylindrical chamber reacted, have a central reaction chamber (2)
Positioned at disc-shaped structure inner ring center, remaining cylindrical chamber is evenly distributed on disc-shaped structure outer ring (1), is used for core with multiple
Sour circular chamber's cross-distribution amplifing reagent preservation and reacted.
2. sample according to claim 1 automatically processes and nucleic acid amplifier, it is characterised in that: the cylindrical center
Reaction chamber (2) includes the nanometer magnetic bead that the rubber plunger and adsorbable nucleic acid that move up and down can be controlled by external force, central reaction
Chamber (2) bottom saving with the cylindrical chamber reacted for sample processing reagent and trying for nucleic acid amplification with outer ring (1)
Agent preservation passes through that microchannel is radial to be connected with the circular chamber reacted respectively, and central reaction chamber (2) bottom is interior
Side wall is enclosed, contains 1 microchannel aperture, central reaction chamber (2) can be made to be connected with other chambers by the horizontal rotation of inner ring
The multiple microchannels connect are opened one by one.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201820196093.3U CN208200972U (en) | 2018-02-05 | 2018-02-05 | A kind of sample automatically processes and nucleic acid amplifier |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201820196093.3U CN208200972U (en) | 2018-02-05 | 2018-02-05 | A kind of sample automatically processes and nucleic acid amplifier |
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| CN208200972U true CN208200972U (en) | 2018-12-07 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108192816A (en) * | 2018-02-05 | 2018-06-22 | 宁波东夏生物科技有限公司 | A kind of sample automatically processes and nucleic acid amplifier and application method |
-
2018
- 2018-02-05 CN CN201820196093.3U patent/CN208200972U/en active Active
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108192816A (en) * | 2018-02-05 | 2018-06-22 | 宁波东夏生物科技有限公司 | A kind of sample automatically processes and nucleic acid amplifier and application method |
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| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20220412 Address after: 314112 room 407, 4 / F, building B, No. 6, dingyang Road, Huimin street, Jiashan County, Jiaxing City, Zhejiang Province Patentee after: Jiaxing Jiasheng Biotechnology Co.,Ltd. Address before: 315040 room 471, 2 building, 66 business building, academician Road, Ningbo high tech Zone, Zhejiang, China Patentee before: NINGBO DONGXIA BIOTECHNOLOGY CO.,LTD. |