CN212357219U - Micro-fluidic chip for nucleic acid amplification - Google Patents
Micro-fluidic chip for nucleic acid amplification Download PDFInfo
- Publication number
- CN212357219U CN212357219U CN202021311252.3U CN202021311252U CN212357219U CN 212357219 U CN212357219 U CN 212357219U CN 202021311252 U CN202021311252 U CN 202021311252U CN 212357219 U CN212357219 U CN 212357219U
- Authority
- CN
- China
- Prior art keywords
- quantitative
- nth
- amplification
- disc
- communicated
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Abstract
The utility model discloses a micro-fluidic chip for nucleic acid amplification, include: a plurality of nested amplification regions for nested amplification of nucleic acid molecules; and a quantitative distribution means for quantitatively dividing and distributing a microfluid containing nucleic acid molecules to each of the nested amplification regions; the dosing device is disposed in a microchannel for providing the microfluidics to the nested amplification zone. The utility model provides the high accuracy that nucleic acid analysis detected.
Description
Technical Field
The utility model belongs to the technical field of biological detection, a micro-fluidic chip for nucleic acid amplification is related to.
Background
When nucleic acid amplification is carried out in a laboratory environment, the problem of nucleic acid pollution interference detection exists more or less, aerosol is easy to diffuse in the air, and then false positive results appear in subsequent sample detection. The micro-fluidic chip has the advantages that the nucleic acid in the environment is difficult to enter the chip to be polluted due to the sealing property of the internal environment of the micro-fluidic chip, the internal pollution condition is effectively prevented due to the independent distribution of the internal cavity of the chip, and the pollution degree of the nucleic acid can be reduced to the lowest degree.
The micro-fluidic chip technology and the nested PCR technology are combined to develop the micro-fluidic chip which can be used for the rapid quantitative detection of the biological sample, a simple, rapid and effective solution is provided for the rapid quantitative detection of nucleic acid molecules in the biological sample, and the method has important significance for the treatment and prognosis of diseases.
SUMMERY OF THE UTILITY MODEL
The utility model aims at providing a micro-fluidic chip for nucleic acid amplification has improved the accuracy that nucleic acid analysis detected.
The utility model provides a micro-fluidic chip for nucleic acid amplification, include:
a plurality of nested amplification regions for nested amplification of nucleic acid molecules; and
a quantitative distribution means for quantitatively dividing and distributing a microfluid containing nucleic acid molecules to each of the nested amplification regions;
the dosing device is disposed in a microchannel for providing the microfluidics to the nested amplification zone.
Preferably, the quantitative distribution device comprises a plurality of quantitative distribution units, and each quantitative distribution unit corresponds to one nested amplification region.
More preferably, the first quantitative distribution unit comprises a movably arranged first quantitative tray, the first quantitative tray is provided with an inner cavity and a liquid inlet and a liquid outlet which are respectively communicated with the inner cavity, the microchannel comprises a first microchannel for sample introduction and a second microchannel arranged between the first quantitative distribution unit and the second quantitative distribution unit, the first quantitative tray is provided with a first position and a second position, when the first quantitative tray is at the first position, the liquid inlet of the first quantitative tray is communicated with the first microchannel, and the liquid outlet is communicated with the second microchannel; when the first quantitative tray is in the second position, the liquid inlet of the first quantitative tray is separated from the first microchannel, and the liquid outlet is communicated with the first nested amplification region.
More preferably, the nth quantitative distribution unit comprises an nth quantitative disc movably arranged, N is a positive integer greater than 1 and less than N, N is the total number of the quantitative distribution units, N is the total number of the nested amplification zones, the nth quantitative disc has an inner cavity, and a liquid inlet and a liquid outlet respectively communicated with the inner cavity, the microchannel further comprises an nth microchannel arranged between the nth quantitative distribution unit and the nth-1 quantitative distribution unit, and an N +1 microchannel arranged between the nth quantitative distribution unit and the N +1 quantitative distribution unit, the nth quantitative disc has a first position and a second position, when the nth quantitative disc is in the first position, the liquid inlet of the nth quantitative disc is communicated with the nth microchannel, and the liquid outlet is communicated with the N +1 microchannel; when the nth quantitative disc is at the second position, the liquid inlet of the nth quantitative disc is separated from the nth microchannel, and the liquid outlet is communicated with the nth nested amplification area.
More preferably, the nth quantitative distribution unit comprises an nth quantitative disc movably arranged, wherein the nth quantitative disc is provided with an inner cavity, and a liquid inlet and a liquid outlet which are respectively communicated with the inner cavity; the quantitative distribution device also comprises an indication area, and an indication area micro-channel is arranged between the Nth quantitative distribution unit and the indication area; the Nth quantitative disc is provided with a first position and a second position, when the Nth quantitative disc is at the first position, the liquid inlet of the Nth quantitative disc is communicated with the Nth micro-channel, and the liquid outlet of the Nth quantitative disc is communicated with the micro-channel of the indicating area; when the Nth quantitative disc is at the second position, the liquid inlet of the Nth quantitative disc is separated from the Nth microchannel, and the liquid outlet is communicated with the Nth nested amplification area.
More preferably, the microfluidic chip comprises a channel layer in which a quantitative disc is rotatably disposed; and/or the quantitative disc is provided with a coil pipe formed by bending for many times, and the inner cavity is formed by a pipe cavity of the coil pipe; and/or each said dosing unit comprises a pneumatic valve, said liquid inlet being in communication with a respective said pneumatic valve when the dosing disc is in the second position; and/or each quantitative disc is synchronized in the first position or in the second position.
Preferably, the nested amplification regions have primers stored therein, and the primers in different nested amplification regions are the same or different; amplification reagents are stored in the nested amplification area, or the microfluidic chip further comprises a liquid supplementing hole which is communicated with the nested amplification area and is used for adding the amplification reagents into the nested amplification area.
Preferably, each of the nested amplification regions is in communication with a vent.
Preferably, the microfluidic chip further comprises a first round amplification area for performing a first round amplification on nucleic acid molecules, and the first round amplification area provides the first round amplified nucleic acid molecules to the quantitative distribution device through the microchannel.
More preferably, the microfluidic chip further comprises a reagent buffer area capable of communicating with the first round amplification area, and the microchannel communicates with the reagent buffer area.
More preferably, the first round amplification region has stored therein amplification reagents required for the first round of nucleic acid amplification.
More preferably, the microfluidic chip further comprises a sample adding hole for adding a sample to the first round amplification region, and a sample adding channel is arranged between the sample adding hole and the first round amplification region.
Further, the microfluidic chip also comprises an air pressure valve arranged in the sample adding channel.
Preferably, the microfluidic chip comprises a channel layer and a cover plate layer fixed above the channel layer, and the nested amplification area is a chamber arranged in the channel layer.
More preferably, a pneumatic valve capable of moving up and down is provided in the cover plate layer.
More preferably, the thickness of the channel layer and the cover plate layer is 1-20 mm respectively.
The utility model adopts the above scheme, compare prior art and have following advantage:
the utility model discloses a micro-fluidic chip can be used for nucleic acid amplification, especially nested PCR amplifys, has not only effectively reduced the nucleic acid pollution degree, combines nested PCR's specificity to amplify simultaneously, has greatly improved the accuracy that nucleic acid analysis detected, and the reagent consumption is low, analysis speed is fast, can also satisfy the demand that many indexes detected, is particularly suitable for the quick quantitative determination of nucleic acid molecule among the biological sample.
Drawings
In order to more clearly illustrate the technical solution of the present invention, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
Fig. 1 is a schematic front view of a microfluidic chip according to an embodiment of the present invention;
FIG. 2 is a schematic structural view of a quantitative plate according to an embodiment of the present invention;
fig. 3 is a schematic illustration of a quantitatively partitioned solution according to an embodiment of the present invention;
fig. 4 is a schematic view of nested amplification zone sample injection according to an embodiment of the present invention.
Wherein the content of the first and second substances,
101. a channel layer; 102. a cover plate layer; 103. positioning holes;
3. a first round amplification region; 31. a sample application channel; 32. a first fluid replenishment hole; 33. a pneumatic valve I;
4. a reagent buffer zone;
5. a nested amplification zone; 51. a vent hole;
6. a dosing device; 60. a dosing unit; 61. a quantitative disc; 611. a coil pipe; 61a, a first quantitative disc; 61b, a second quantitative disc; 62a, a first microchannel; 62b, a second microchannel; 62c, a third microchannel; 63. an indication area; 64. an indicator zone microchannel; 65a, a pneumatic valve II; 65b, a pneumatic valve III; 65c, a pneumatic valve IV; 65d, a pneumatic valve V; 65e, a pneumatic valve VI; 65f, a pneumatic valve VII; 65g and a pneumatic valve XIII.
Detailed Description
The following detailed description of the preferred embodiments of the invention, taken in conjunction with the accompanying drawings, enables the advantages and features of the invention to be more readily understood by those skilled in the art. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto.
As used in this specification and the appended claims, the terms "comprises" and "comprising" are intended to only encompass the explicitly identified steps and elements, which do not constitute an exclusive list, and that a method or apparatus may include other steps or elements. As used herein, the term "and/or" includes any combination of one or more of the associated listed items.
It should be noted that, unless otherwise specified, when a feature is referred to as being "fixed" or "connected" to another feature, it may be directly fixed or connected to the other feature or indirectly fixed or connected to the other feature. Furthermore, the description of the upper, lower, left, right, etc. used in the present invention is only relative to the mutual positional relationship of the components of the present invention in the drawings.
It is further understood that the use of "a plurality" in this disclosure means two or more, as other terms are analogous. "and/or" describes the association relationship of the associated objects, meaning that there may be three relationships, e.g., a and/or B, which may mean: a exists alone, A and B exist simultaneously, and B exists alone.
The present example provides a microfluidic chip for nucleic acid amplification. Referring to fig. 1, the microfluidic chip mainly comprises a first round amplification area 3 for performing a first round amplification on nucleic acid molecules and a nested amplification area 5 for performing nested amplification on the nucleic acid molecules after the first round amplification, wherein the nested amplification area 5 can be communicated with the first round amplification area 3. Wherein, the number of the nested amplification regions 5 is a plurality, and each nested amplification region 5 can be communicated with the first round amplification region 3 through a micro-channel. The microfluidic chip further comprises a quantitative distribution device 6 for quantitatively dividing and distributing the microfluid in the first round amplification region 3 to each nested amplification region 5, wherein the microfluid can be divided uniformly or non-uniformly. In this embodiment, the division is uniform.
The first-round amplification region 3 stores therein amplification reagents required for the first-round nucleic acid amplification, and the amplification reagents may be pre-stored in a liquid form in the first-round amplification region 3. The amplification reagent comprises various primers, dNTP, biological enzyme and other reagents required by nucleic acid amplification, can perform specific amplification on nucleic acid molecules, and meets the detection requirement of biological samples.
The microfluidic chip further comprises a sample adding hole 32 for adding a biological sample containing nucleic acid molecules into the first round amplification area 3, and a sample adding channel 31 is arranged between the sample adding hole 32 and the first round amplification area 3. The sample feeding channel 31 is provided with a pneumatic valve I33.
The microfluidic chip further comprises a reagent buffer zone 4 arranged between the first round amplification zone 3 and the nested amplification zone 5. A reagent buffer zone 4 for eliminating the functional area of air bubbles in the first round of PCR amplification products; can also be used as a functional area for mixing the first round PCR amplification product and the nested PCR amplification reagent.
Primers are stored in each nested amplification region 5 respectively, and the primers in different nested amplification regions 5 are the same or different, because the second pair of primers has higher specificity and must be stored in corresponding chambers separately; the primers can be pre-stored in the nested amplification area 5 in a freeze-dried powder form and can be melted after meeting the solution. In this embodiment, the primers in each nested amplification region 5 are different from each other, thereby realizing multi-index detection. The nested amplification area 5 is also stored with amplification reagents for the second round of nucleic acid amplification, or the microfluidic chip further comprises a liquid replenishing hole for adding the amplification reagents into the nested amplification area 5. A segment of DNA within the first round of PCR products can be amplified in nested amplification region 5, thereby increasing the specificity of the reaction. Nested amplification region 5 can be selected to store the second primer pair, dNTP, various biological enzymes, etc. required for nucleic acid amplification, and if only the second primer pair is stored, but no other nested PCR amplification reagents are stored, the required reagents can be added from second replenisher well 42. The nested amplification region 5 also allows for the presence of a detection region that is a product of nucleic acid amplification. Each nested amplification area 5 is respectively communicated with a vent hole 51, and the vent hole 51 is a position on the microfluidic chip communicated with the external atmosphere and is used for ensuring the balance of the internal and external air pressures of the chip when the microfluid distributed by the quantitative distribution device 6 flows into the nested amplification areas 5.
Referring to FIGS. 2 to 4, the quantitative distribution means 6 is provided in a microchannel for supplying microfluidics to the nested amplification regions 5. The quantitative distribution device 6 comprises a plurality of quantitative distribution units 60, and each quantitative distribution unit 60 corresponds to one nested amplification region 5. Each quantitative distribution unit 60 comprises a quantitative disc 61, and the quantitative disc 61 is provided with an inner cavity and a liquid inlet and a liquid outlet which are respectively communicated with the inner cavity. The quantitative trays 61 can be moved or rotated to have a first position in which the liquid inlet is in direct or indirect communication with the reagent buffer 4 and the liquid outlet is in communication with the liquid inlet or indicator 63 of the next quantitative tray 61, so that the microfluid from the first round of amplification zones 3 or reagent buffer 4 flows into the inner chamber; when microfluid is distributed in the inner cavity of each quantitative disc 61, the quantitative discs 61 move for a certain distance or rotate for a certain angle, thereby being switched to a second position; in the second position, the liquid inlet is closed or connected with a pneumatic valve (pneumatic valve II65 a, pneumatic valve III65 b, pneumatic valve IV 65c, pneumatic valve V65 d, pneumatic valve VI 65e, pneumatic valve VII 65f), the liquid outlet is connected with the corresponding nested amplification zone 5, and the microfluid in the inner cavity is injected into the nested amplification zone 5. The above-mentioned indication area 63 serves as an area for identifying a process of quantitatively dividing the solution for terminating the operation of dividing the solution. The dosing unit 60 is illustrated below.
The quantitative distribution unit 60 directly communicating with the reagent buffer 4 will be referred to as a first quantitative distribution unit. The first quantitative distribution unit comprises a first quantitative disc 61a which is movably arranged, and the first quantitative disc 61a is provided with an inner cavity, a liquid inlet and a liquid outlet which are respectively communicated with the inner cavity. A first microchannel 62a is provided between the first round amplification region 3 and the first quantitative distribution unit 60, and a second microchannel 62b is provided between the first quantitative distribution unit and the second quantitative distribution unit. The first quantitative disc 61a has a first position and a second position, when the first quantitative disc 61a is in the first position, the liquid inlet of the first quantitative disc 61a is communicated with the first micro-channel 62a, and the liquid outlet is communicated with the second micro-channel 62b (as shown in FIG. 3); when the first quantitative tray 61a is in the second position, the inlet of the first quantitative tray 61a is separated from the first microchannel 62a, and the outlet is in communication with the first nested amplification sections 5 (see FIG. 4).
The metering unit 60 farthest from the reagent buffer 4 is referred to as the Nth metering unit. The Nth quantitative distribution unit comprises an Nth quantitative disc which is movably arranged, and the Nth quantitative disc is provided with an inner cavity, a liquid inlet and a liquid outlet which are respectively communicated with the inner cavity. The dosing device 6 further comprises the above mentioned indicator area 63, and an indicator area microchannel 64 is arranged between the nth dosing unit and the indicator area 63. The Nth quantitative disc is provided with a first position and a second position, when the Nth quantitative disc is at the first position, the liquid inlet of the Nth quantitative disc is communicated with the Nth micro-channel, and the liquid outlet of the Nth quantitative disc is communicated with the micro-channel of the indicating area; when the Nth quantitative disc is at the second position, the liquid inlet of the Nth quantitative disc is separated from the Nth microchannel, and the liquid outlet is communicated with the Nth nested amplification area.
Let any one of the first and nth metering units be denoted as the nth metering unit, N is a positive integer greater than 1 and less than N, and N is the total number of metering units 60 or the total number of nested amplification regions 5. The nth quantitative distribution unit comprises an nth quantitative disc which is movably arranged, and the nth quantitative disc is provided with an inner cavity, a liquid inlet and a liquid outlet which are respectively communicated with the inner cavity. An nth microchannel is arranged between the nth quantitative distribution unit and the (n-1) th quantitative distribution unit, and an (n + 1) th microchannel is arranged between the nth quantitative distribution unit and the (n + 1) th quantitative distribution unit. The nth quantitative disc is provided with a first position and a second position, when the nth quantitative disc is at the first position, the liquid inlet of the nth quantitative disc is communicated with the nth microchannel, and the liquid outlet of the nth quantitative disc is communicated with the (n + 1) th microchannel; when the nth quantitative disc is at the second position, the liquid inlet of the nth quantitative disc is separated from the nth microchannel, and the liquid outlet is communicated with the nth nested amplification area 5.
Specifically, in this embodiment, the number of the nested amplification regions 5 and the number of the quantitative distribution units 60 are all six and are sequentially arranged from left to right in the order of the first to the sixth. N is 6, and the nth quantitative distribution unit is the sixth quantitative distribution unit. When n is 2, the nth quantitative distribution unit 60 is the second quantitative distribution unit. The second quantitative distribution unit comprises a second quantitative disc 61b which is movably arranged, the second quantitative disc 61b is provided with an inner cavity, a liquid inlet and a liquid outlet which are respectively communicated with the inner cavity, a third micro-channel 62c is arranged between the second quantitative distribution unit and the third quantitative distribution unit, the second quantitative disc 61b is provided with a first position and a second position, when the second quantitative disc 61b is at the first position, the liquid inlet of the second quantitative disc 61b is communicated with the second micro-channel 62b, and the liquid outlet is communicated with the third micro-channel 62 c; when the second quantitative tray 61b is in the second position, the inlet of the second quantitative tray 61b is separated from the second microchannel 62b, and the outlet is connected to the second nested amplification section 5. And the conditions that the value of n is 3, 4 and 5 are analogized in sequence, namely the structures of the third, the fourth and the fifth quantitative distribution units are similar to the structure of the second quantitative distribution unit. It should be noted that the microchannels (the second to sixth microchannels described above) between two adjacent quantitative disks 61 are S-shaped.
The embodiment further provides a specific structure of the quantitative disc. As shown in fig. 2, the quantitative plate 61 has a coil 611 formed by multiple bending, and the inner cavity is formed by the lumen of the coil 611. The respective quantitative discs 61 are synchronized in the first position (as shown in fig. 3) or in the second position. Each of the quantitative distribution units 60 includes pneumatic valves (pneumatic valve II65 a, pneumatic valve III65 b, pneumatic valve IV 65c, pneumatic valve V65 d, pneumatic valve VI 65e, pneumatic valve VII 65f) and the liquid inlets communicate with the respective pneumatic valves when the quantitative trays 61 are in the second position to inject the microfluids distributed into the respective quantitative trays 61 into the respective nested amplification zones 5. The above-mentioned quantitative distribution means 6 further comprises a pneumatic valve VIII65g for regulating the pressure in the indication section 63 for allowing the microfluid in the reagent buffer section 4 to flow into the respective quantitative disks 61 in sequence.
The microfluidic chip comprises a channel layer and a cover plate layer 102 fixed above the channel layer, and the thickness of the channel layer and the thickness of the cover plate layer 102 are respectively 1-20 mm. The first round amplification region 3, the reagent buffer region 4 and the nested amplification region 5 are chambers respectively arranged in the channel layer. The microchannels (including the first to nth microchannels, the sample loading channel, etc.) for communicating different chambers are arranged in the channel layer. The quantitative disks 61 are rotatably disposed in the channel layer about their central axes, and each quantitative disk 61 is connected to a quantitative disk driving member rotatably disposed in the cover layer 102. The pneumatic valves (I to VIII) are disposed in the cover plate layer 102 so as to be movable up and down; the charging piston 14 and the opening/closing piston 15 are provided on the cover plate layer 102 so as to be movable up and down. On one hand, after the cover plate layer 102 is bonded with the channel layer, a closed cavity is established; on the other hand, the pneumatic valve, the liquid adding piston 14 and the switch piston 15 are provided with movable strokes, and the functions of quantitative driving and controlling of the fluid are realized.
The channel layer and the cover plate layer 102 are respectively provided with a positioning hole 103 which is matched with each other and used for bonding and positioning the channel layer and the cover plate layer 102. The positioning holes 103 are respectively positioned at four corners of the microfluidic chip.
The channel layer and cover plate layer 102 may be made of a polymer such as PS, PMMA, PDMA, PC, or glass or metal.
In this embodiment, the fluid driving method is mainly air pressure driving, that is, the microfluid on the microfluidic chip is driven to a specific position by air pressure difference, specifically, a liquid adding piston or an air pressure valve is adopted, specifically, a columnar piston is adopted as the air pressure valve, and is located in the middle or at one end of the microchannel. The initial position of the piston can be corrected to a certain position to drive the solution quantitatively, and the solution can also be driven quantitatively by an external precise injection pump.
The embodiment also provides a method for detecting nucleic acid, which adopts the microfluidic chip to detect, and the specific process is described as follows.
Step one, adding a sample to the microfluidic chip through the sample adding hole 32, and performing first round PCR amplification by changing the temperature of the nucleic acid first round amplification area 3.
And step two, after the first round of PCR amplification is finished, driving the microfluid of the first round of amplification region 3 to six quantitative discs 61, and quantitatively dividing the solution of the first round of nucleic acid amplification region 3. The operation mode is two types: one mode is that the air pressure valve VIII is removed, the position of the air pressure valve VIII is changed into an air vent, then the air pressure valve I is pressed to the bottom, and then microfluid of the first round amplification area 3 sequentially flows through each quantitative disc 61, so that the function of quantitatively dividing the solution is realized; in another mode, the air pressure valve I is pressed to the bottom at a certain speed, the air pressure valve VIII is lifted at the same speed, and the operation of quantitatively dividing the solution is stopped when the solution appears in the indicating area 63, so that the air pressure balance between the inside and the outside of the microfluidic chip is kept.
And step three, after the operation of quantitatively dividing the solution is finished, rotating the quantitative discs 61 counterclockwise by a certain angle to enable each quantitative disc 61 to be respectively communicated with the liquid path between the corresponding air pressure valve and the nested amplification area 5. When the air pressure valves II-VII are pressed to the bottom, the solutions in the coil 611 of the quantitative tray 61 enter the nested amplification area 5 respectively, and are mixed and reacted with the second pair of primers, dNTP, biological enzyme and other reagents which are pre-stored in the nested amplification area 5 and are needed by nucleic acid amplification, and nested PCR amplification is carried out by changing the temperature of the nested amplification area 5. And after the nested PCR amplification is finished, reading the fluorescent signal of the nested amplification area 5 by a fluorescent detection technology to finish the detection process.
Application example 1
By adopting the method for nucleic acid amplification, different primers are respectively added to six nested amplification regions 5, and only 1 primer is added to each nested amplification region 5, so that the detection requirements of 6 indexes in a biological sample can be realized in a nested PCR mode.
Application example 2
By adopting the method for detecting nucleic acid, different primers are respectively added in six nested amplification areas 5, m (m is more than 1) primers are added in each nested amplification area 5, the types of the primers are different, and the primers can be marked by fluorescent groups with different colors, so that the detection requirements of 6m indexes can be realized, the flux of nucleic acid analysis and detection is greatly improved, and the detection time of multiple indexes is shortened.
Application example 3
By adopting the method for detecting nucleic acid, the microfluidic chip with seven nested amplification areas 5 and seven quantitative distribution units 60 is adopted, different primers are respectively added in the seven nested amplification areas 5, m (m is more than 1) primers are added in each nested amplification area 5, the types of the primers are different, and the primers can be marked by fluorescent groups with different colors, so that the detection requirement of 7m indexes can be realized.
The micro-fluidic chip is used for nested PCR amplification, not only effectively reduces the degree of nucleic acid pollution, but also greatly improves the accuracy of nucleic acid analysis and detection by combining the specific amplification of the nested PCR.
In addition, the structure of the microfluidic chip can be properly adjusted according to the requirement of detecting the index quantity, for example, 7n indexes in a biological sample need to be detected, and the number of the quantitative disc 61 and the nucleic acid second-round amplification area can be enlarged to 7. Scaling up or down the number of quantification discs 61 and second round nucleic acid amplification regions (greater than 1) may satisfy the detection requirements for any scalar.
The above embodiments are only for illustrating the technical concept and features of the present invention, and are preferred embodiments, which are intended to enable persons skilled in the art to understand the contents of the present invention and to implement the present invention, and thus, the protection scope of the present invention cannot be limited thereby. All equivalent changes or modifications made according to the principles of the present invention are intended to be covered by the scope of the present invention.
Claims (10)
1. A microfluidic chip for nucleic acid amplification, comprising:
a plurality of nested amplification regions for nested amplification of nucleic acid molecules; and
a quantitative distribution means for quantitatively dividing and distributing a microfluid containing nucleic acid molecules to each of the nested amplification regions;
the dosing device is disposed in a microchannel for providing the microfluidics to the nested amplification zone.
2. The microfluidic chip of claim 1, wherein: the quantitative distribution device comprises a plurality of quantitative distribution units, and each quantitative distribution unit corresponds to one nested amplification region.
3. The microfluidic chip of claim 2, wherein: the first quantitative distribution unit comprises a first quantitative disc which is movably arranged, the first quantitative disc is provided with an inner cavity, a liquid inlet and a liquid outlet which are respectively communicated with the inner cavity, the micro channel comprises a first micro channel for sample introduction and a second micro channel which is arranged between the first quantitative distribution unit and the second quantitative distribution unit, the first quantitative disc is provided with a first position and a second position, when the first quantitative disc is at the first position, the liquid inlet of the first quantitative disc is communicated with the first micro channel, and the liquid outlet is communicated with the second micro channel; when the first quantitative tray is in the second position, the liquid inlet of the first quantitative tray is separated from the first microchannel, and the liquid outlet is communicated with the first nested amplification region; and/or the nth quantitative distribution unit comprises an nth quantitative disc which is movably arranged, wherein N is a positive integer which is more than 1 and less than N, N is the total number of the quantitative distribution unit which is the total number of the nested amplification areas, the nth quantitative disc is provided with an inner cavity, a liquid inlet and a liquid outlet which are respectively communicated with the inner cavity, the microchannel further comprises an nth microchannel which is arranged between the nth quantitative distribution unit and the nth-1 quantitative distribution unit and an N +1 microchannel which is arranged between the nth quantitative distribution unit and the N +1 quantitative distribution unit, the nth quantitative disc is provided with a first position and a second position, when the nth quantitative disc is at the first position, the liquid inlet of the nth quantitative disc is communicated with the nth microchannel, and the liquid outlet is communicated with the N +1 microchannel; when the nth quantitative disc is at the second position, the liquid inlet of the nth quantitative disc is separated from the nth microchannel, and the liquid outlet is communicated with the nth nested amplification region; and/or the Nth quantitative distribution unit comprises an Nth quantitative disc which is movably arranged and is provided with an inner cavity, a liquid inlet and a liquid outlet which are respectively communicated with the inner cavity; the quantitative distribution device also comprises an indication area, and an indication area micro-channel is arranged between the Nth quantitative distribution unit and the indication area; the Nth quantitative disc is provided with a first position and a second position, when the Nth quantitative disc is at the first position, the liquid inlet of the Nth quantitative disc is communicated with the Nth micro-channel, and the liquid outlet of the Nth quantitative disc is communicated with the micro-channel of the indicating area; when the Nth quantitative disc is at the second position, the liquid inlet of the Nth quantitative disc is separated from the Nth microchannel, and the liquid outlet is communicated with the Nth nested amplification area.
4. The microfluidic chip according to claim 3, wherein: the microfluidic chip comprises a channel layer, and the quantitative disc is rotatably arranged in the channel layer; and/or the quantitative disc is provided with a coil pipe formed by bending for many times, and the inner cavity is formed by a pipe cavity of the coil pipe; and/or each said dosing unit comprises a pneumatic valve, said liquid inlet being in communication with a respective said pneumatic valve when the dosing disc is in the second position; and/or each quantitative disc is synchronized in the first position or in the second position.
5. The microfluidic chip of claim 1, wherein: primers are stored in the nested amplification regions, and the primers in different nested amplification regions are the same or different; and/or amplification reagents are stored in the nested amplification area, or the microfluidic chip further comprises a liquid supplementing hole which is communicated with the nested amplification area and is used for adding the amplification reagents into the nested amplification area; and/or each nested amplification region is respectively communicated with a vent hole.
6. The microfluidic chip of claim 1, wherein: the microfluidic chip further comprises a first round amplification area for performing first round amplification on nucleic acid molecules, and the first round amplification area provides the nucleic acid molecules after the first round amplification to the quantitative distribution device through the micro-channel.
7. The microfluidic chip according to claim 6, wherein: the microfluidic chip also comprises a reagent buffer area which can be communicated with the first round amplification area, and the microchannel is communicated with the reagent buffer area.
8. The microfluidic chip according to claim 7, wherein: the microfluidic chip further comprises a sample adding hole for adding a sample into the first round amplification area, and a sample adding channel is arranged between the sample adding hole and the first round amplification area.
9. The microfluidic chip of claim 8, wherein: the microfluidic chip also comprises an air pressure valve arranged in the sample adding channel.
10. The microfluidic chip of claim 1, wherein: the micro-fluidic chip comprises a channel layer and a cover plate layer fixed above the channel layer, and the nested amplification area is a chamber arranged in the channel layer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202021311252.3U CN212357219U (en) | 2020-07-07 | 2020-07-07 | Micro-fluidic chip for nucleic acid amplification |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202021311252.3U CN212357219U (en) | 2020-07-07 | 2020-07-07 | Micro-fluidic chip for nucleic acid amplification |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN212357219U true CN212357219U (en) | 2021-01-15 |
Family
ID=74152302
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202021311252.3U Active CN212357219U (en) | 2020-07-07 | 2020-07-07 | Micro-fluidic chip for nucleic acid amplification |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN212357219U (en) |
-
2020
- 2020-07-07 CN CN202021311252.3U patent/CN212357219U/en active Active
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111592971B (en) | Microfluidic chip and method for detecting nucleic acid | |
| JP5049404B2 (en) | Fluid handling and transfer methods using interconnected multi-chamber devices | |
| US9260750B2 (en) | Mobile rapid test system for nucleic acid analysis | |
| US6818435B2 (en) | Microfluidics devices and methods for performing cell based assays | |
| US7906318B2 (en) | Testing microreactor, testing device and testing method | |
| US9677133B2 (en) | Biological chip hybridization system | |
| EP1181541B1 (en) | Apparatus and method for sample analysis | |
| US6821771B2 (en) | Device for thermo-dependent chain reaction amplification of target nucleic acid sequences, measured in real-time | |
| US20050196779A1 (en) | Self-contained microfluidic biochip and apparatus | |
| US20050221281A1 (en) | Self-contained microfluidic biochip and apparatus | |
| US20100252128A1 (en) | Microfluidic device | |
| US20050136545A1 (en) | Microfluidics devices and methods for performing based assays | |
| WO2010114858A1 (en) | Reservoir-buffered mixers and remote valve switching for microfluidic devices | |
| Kao et al. | A fluorescence in situ hybridization (FISH) microfluidic platform for detection of HER2 amplification in cancer cells | |
| CN111592972A (en) | Micro-fluidic chip and method for nucleic acid amplification | |
| CN111621417B (en) | Microfluidic chip for biological sample processing and application method thereof | |
| EP2307882B1 (en) | System and method to prevent cross-contamination in assays performed in a microfludic channel | |
| CN212357219U (en) | Micro-fluidic chip for nucleic acid amplification | |
| CN110616144A (en) | Liquid drop digital PCR chip and use method thereof | |
| CN115651807B (en) | Nucleic acid detection chip and nucleic acid detection method | |
| CN212247004U (en) | Micro-fluidic chip for nucleic acid detection | |
| CN205329004U (en) | Dull and stereotyped isothermal nucleic acid increase chip | |
| CN105400692B (en) | Isothermal nucleic acid amplification device and isothermal nucleic acid amplification experimental technique | |
| CN216738284U (en) | Loop-mediated isothermal amplification chip | |
| CN117181326A (en) | Flow channel structure, centrifugal microfluidic chip and use method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant |