CN208829684U - Digital PCR system - Google Patents

Abstract

The utility model provides a digital PCR system, this digital PCR system include that at least one liquid drop forms subassembly and liquid drop orifice subassembly, the liquid drop forms the subassembly and includes at least one liquid drop collecting vat, liquid drop orifice subassembly connect in liquid drop forms subassembly below, including a plurality of liquid drop orifices, the liquid drop orifice with liquid drop collecting vat intercommunication, just be equipped with vaporization part in the liquid drop orifice, be used for making digital PCR solution liquid layer vaporization in the liquid drop orifice is pushed fast in liquid drop forming oil in the liquid drop collecting vat to form digital PCR liquid drop. The utility model discloses use the hot bubble technique to carry out high-speed digital PCR liquid drop formation, can realize being greater than 1000 liquid drop formation speed per second to have efficient digital PCR oil utilization rate.

Description

A kind of digital pcr system

Technical field

The utility model belongs to biomedicine field, especially disease detection field, is related to a kind of digital pcr system.

Background technique

Polymerase chain reaction (polymerase chain reaction, PCR) proposes have 20 years, phase so far Between PCR have evolved into a key technology and routine techniques of molecular biology field, greatly pushed life science each The development in a field.The especially later period nineties, American AB I company release real-time fluorescence quantitative PCR (real time PCR, QPCR) technology and Related product are even more by PCR by synthesizing in vitro and qualitative/half-quantitative detection technology develops into a kind of Gao Ling The gene analysis technique of quick, high specific and accurate quantification.

Although qPCR technology has been used to own in addition to wound and deficiency disease by the rapid development of more than ten years The diagnosis of disease still influences many because being known as of its amplification efficiency, it cannot be guaranteed that in reaction process during PCR amplification Middle amplification efficiency remain unchanged the amplification efficiency between actual sample and standard sample and different samples be it is identical, thus It is directed at the basis that its quantitative analysis is relied on --- cycle threshold (CT) is not constant between.Therefore the quantitative of qPCR is Still it is impossible to meet the requirements of molecular biology quantitative analysis for " relative quantification ", accuracy and reproducibility.

At the end of the 20th century, Vogelstein etc. proposes the concept of digital pcr (digital PCR, dPCR), by by a sample Originally it is divided into tens to tens of thousands of parts, is assigned to different reaction members, each unit includes the target molecule of one or more copies (DNA profiling) carries out PCR amplification to target molecule respectively in each reaction member, to each reaction member after amplification Fluorescence signal carry out statistical analysis.Unlike qPCR, digital pcr is independent of CT value, therefore not by amplification efficiency It influences, by directly counting or Poisson distribution formula calculates the mean concentration (content) of each reaction member after amplification, Can be by control errors within 5%, digital pcr can not need reference standard sample and standard curve to realize absolute quantitation Analysis.

Digital pcr (can also claim single-molecule PCR) generally comprises two parts content, i.e. PCR amplification and fluorescence signal analysis.? The PCR amplification stage, different from traditional technology, digital pcr, which is generally required, is diluted to single molecules level for sample, and is evenly distributed to Tens are reacted into tens of thousands of a units.Real-time fluorescence method for measuring, digital pcr are carried out to each circulation different from qPCR Technology is to be acquired after amplification to the fluorescence signal of each reaction member.Finally by directly counting or Poisson distribution The original concentration or content of sample is calculated in formula.

Since digital pcr is that a kind of nucleic acid molecules absolute quantitation technology compared to qPCR can directly count DNA molecular Number, be the absolute quantitation to initial sample, be therefore particularly suitable for the application field that cannot be differentiated very well by CT value, example Such as copy number variation, abrupt climatic change, gene relative expression research (such as allele imbalance express), two generation sequencing results are tested Card, miRNA expression analysis, unicellular gene expression analysis etc..

At present there are mainly three types of digital pcr technologies on the market.One is by using the oil of flowing in particular instrument The PCR solution for cutting off water phase forms drop, and PCR and detection are then completed in two other instruments；One is by by PCR Solution is distributed on the silicon wafer hollowed out, then carries out being detected in PCR and an other instrument in particular instrument；Finally One is liquid is formed drop by narrow Channeling implantation cavity on a kind of instrument, and PCR is completed, then at another Detection is completed in instrument.However, the droplet formation speed or flux of three kinds of current methods are each restricted.In addition, above-mentioned three The more large-scale instruments of dependence of kind technology without exception.This not only increases the cost of instrument purchased, and limits digital pcr It is widely used；And increase the complexity of experimental implementation.

Therefore, how digital pcr droplet formation technology, the liquid of a kind of high speed greater than 1000 drops of formation per second are provided Drop forms the situ PCR integrated with PCR temperature control and detecting instrument, efficient digital pcr oil utilization rate method, becomes ability A field technique personnel important technological problems urgently to be resolved.

Utility model content

In view of the foregoing deficiencies of prior art, the purpose of this utility model is to provide a kind of digital pcr systems, use In solving the problems, such as that droplet formation speed is slow in the prior art, flux is small, complicated for operation, the oily utilization rate of PCR is low.

In order to achieve the above objects and other related objects, the utility model provides a kind of digital pcr system, comprising:

At least one droplet formation component, including at least one drop collecting tank；

Drop spray orifice component is connected to below the droplet formation component, including several drop spray orifices, the drop spray orifice It is open, and extends toward the lower surface direction of the drop spray orifice component, but do not run through from the upper surface of the drop spray orifice component The lower surface of the drop spray orifice component, the drop spray orifice are connected to the drop collecting tank, and are set in the drop spray orifice There is vaporization member, is collected for vaporizing the digital pcr aqueous liquids layer in the drop spray orifice and being quickly pushed into the drop In droplet formation oil in slot, to form digital pcr drop.

Optionally, the drop spray orifice component includes thermal printing chip.

Optionally, the vaporization member is set to bottom surface or the side wall of the drop spray orifice.

Optionally, the opening shape of the drop spray orifice includes round, ellipse, any one in polygon.

Optionally, the vaporization member includes heating element, makes its vapour by heating the digital pcr aqueous liquids layer Change.

Optionally, the heating element includes at least one layer of metal layer.

Optionally, the drop collects trench bottom and is equipped with through slot, and the through slot exposes multiple drop spray orifices.

Optionally, the PCR system further includes at least one PCR reagent chamber for being used to store digital pcr solution, described Runner is equipped in drop spray orifice component, the drop spray orifice passes through the runner and the PCR reagent chamber.

Optionally, the runner includes at least one sprue and a plurality of branch flow passage that connect with the sprue, each The drop spray orifice is connect with a branch flow passage respectively.

Optionally, the digital pcr system further includes pedestal, and the PCR reagent chamber is set in the pedestal, described Drop spray orifice component is connected to above the pedestal.

Optionally, the pedestal includes the first base assembly and the second base assembly, and the PCR reagent chamber includes PCR Reagent upper chamber and PCR reagent lower chambers, the PCR reagent upper chamber are open from the upper surface of first base assembly, and Through the lower surface of first base assembly, the PCR reagent lower chambers are open from the upper surface of second base assembly, And extend to the lower surface direction of second base assembly, but do not run through the lower surface of second base assembly, the PCR Reagent upper chamber is connected to PCR reagent lower chambers and part is folded.

Optionally, the lower surface of second base assembly is equipped at least one digital pcr solution injection hole, the number PCR solution injection hole is connected to the PCR reagent lower chambers.

Optionally, the PCR reagent lower chambers include first end and second end, and the digital pcr solution injection hole is in institute It states and is connected at first end with the PCR reagent lower chambers, the PCR reagent lower chambers are tried at the second end with the PCR The connection of agent upper chamber, the size of the PCR reagent lower chambers are gradually increased first from the first end to second extreme direction, Then it is gradually reduced.

Optionally, the lower surface of second base assembly is equipped at least one gas vent, and the gas vent passes through air flue It is connected to the PCR reagent upper chamber, the air flue is open from the upper surface of second base assembly, and to second base The lower surface direction of holder assembly extends, but does not run through the lower surface of second base assembly.

Optionally, first base assembly is fixed on above second base assembly by adhesive means.

Optionally, the digital pcr system further includes flexible circuit board, and the flexible circuit board is connected on the pedestal It is square, the through-hole for accommodating the drop spray orifice component is equipped in the flexible circuit board, and the flexible circuit board surface is set There are several first connection weld pads and several second connection weld pads, the drop spray orifice component to connect weldering with described first by conducting wire Pad connection.

Optionally, the flexible circuit board is connect by adhesive means with the pedestal, and the base-plates surface is equipped with and is used for Prevent glue from flowing to the channel on the drop spray orifice component.

Optionally, in the flexible circuit board be equipped at least two positional punches, the base-plates surface be equipped with it is described fixed The corresponding positioning protrusion of position punch position.

Optionally, the digital pcr system further includes a controller, and the controller includes controller housing and is located at institute The controller circuit board in controller housing is stated, the controller housing has the supporting part for placing the pedestal, described Supporting part surface is equipped with several circuit connection conductive pins connecting with the controller circuitry connecting plate, and the circuit connection is conductive Needle is corresponding with the second connection position of weld pad.

Optionally, one end of the pedestal is provided at least one limiting slot, and the controller housing is provided at least one A locating part corresponding with the limiting slot.

Optionally, the pedestal is provided with a limiting through hole, and the limiting through hole runs through the front and the back side of the pedestal, The controller housing is provided with locating part corresponding with the limiting through hole.

As described above, the digital pcr system of the utility model, has the advantages that

(1) the utility model carries out high-speed figure PCR droplet formation using thermal technology, and the quick formation of drop depends on Vaporization member in drop spray orifice vaporizes the transient heating of nanometer grade thickness liquid level, thus by the number in drop spray orifice PCR solution is quickly pushed into droplet formation oil to form digital pcr drop, compared to the shape of 100 drops each second on the market At speed, the droplet formation technology in the utility model may be implemented to be greater than 1000 droplet formation speed per second.

(2) compared to it is oily mutually and the water phase associated movement method that generates drop, oil in the technical solution of the utility model It is mutually static, therefore the consumption of oily phase is greatly reduced, and reduces 50% or so oily phase dosage.

Detailed description of the invention

Fig. 1 a is shown as the schematic perspective view of the digital pcr system of the utility model.

Fig. 1 b is shown as the top view of the digital pcr system of the utility model.

Fig. 1 c is shown as the bottom view of the digital pcr system of the utility model.

Fig. 1 d- Fig. 1 g is shown as the side view of the digital pcr system of the utility model.

Fig. 2 is shown as the decomposition texture schematic diagram of the digital pcr system of the utility model.

Fig. 3 is shown as the schematic perspective view of drop spray orifice component in the digital pcr system of the utility model.

Fig. 4 is shown as the partial cutaway view of drop spray orifice component in the digital pcr system of the utility model.

Fig. 5 a is shown as the schematic diagram of front three-dimensional structure of droplet formation component in the digital pcr system of the utility model.

Fig. 5 b is shown as the back side dimensional structure diagram of droplet formation component in the digital pcr system of the utility model.

Fig. 5 c is shown as the top view of droplet formation component in the digital pcr system of the utility model.

Fig. 5 d is shown as the bottom view of droplet formation component in the digital pcr system of the utility model.

Fig. 5 e- Fig. 5 h is shown as the side view of droplet formation component in the digital pcr system of the utility model.

Fig. 6 a is shown as the schematic perspective view of pedestal in the digital pcr system of the utility model.

Fig. 6 b is shown as the top view of pedestal in the digital pcr system of the utility model.

Fig. 6 c is shown as the bottom view of pedestal in the digital pcr system of the utility model.

Fig. 6 d- Fig. 6 g is shown as the side view of pedestal in the digital pcr system of the utility model.

Fig. 7 is shown as the partial top view of pedestal in the digital pcr system of the utility model.

Fig. 8 is shown as the schematic diagram of front three-dimensional structure of the second base assembly in the digital pcr system of the utility model.

Fig. 9 is shown as the back side dimensional structure diagram of the second base assembly in the digital pcr system of the utility model.

Figure 10 a is shown as drop spray orifice component in the digital pcr system of the utility model and is being connected to flexible circuit board just Face schematic perspective view.

Figure 10 b is shown as the back that drop spray orifice component in the digital pcr system of the utility model is connected to flexible circuit board Face structural schematic diagram.

Figure 10 c is shown as drop spray orifice component in the digital pcr system of the utility model and is connected to bowing for flexible circuit board View.

Figure 10 d is shown as drop spray orifice component in the digital pcr system of the utility model and is connected to facing upward for flexible circuit board View.

Figure 10 e- Figure 10 h is shown as drop spray orifice component in the digital pcr system of the utility model and is connected to soft route The side view of plate.

Figure 11 is shown as the schematic perspective view of controller in the digital pcr system of the utility model.

Figure 12 is shown as the optical microscope of the digital pcr drop formed using the digital pcr system of the utility model.

Component label instructions

1 droplet formation component

2 drop collecting tanks

3 drop spray orifice components

4 drop spray orifices

5 vaporization members

6 through slots

7 limiting through hole

8 air flues

9 sprues

10 branch flow passages

11 pedestals

12 first base assemblies

13 second base assemblies

14 PCR reagent upper chamber

15 PCR reagent lower chambers

16 digital pcr solution injection holes

17 gas vents

18 flexible circuit board

19 through-holes

20 second connection weld pads

21 channels

22 positional punches

23 positioning protrusion

24 controllers

25 controller housings

26 supporting parts

27 circuit connection conductive pins

28 limiting slots

29,30 locating part

31 boss

Specific embodiment

Illustrate the embodiments of the present invention below by way of specific specific example, those skilled in the art can be by this theory Content disclosed by bright book understands other advantages and effect of the utility model easily.The utility model can also be by addition Different specific embodiments are embodied or practiced, and the various details in this specification can also be based on different viewpoints and answer With carrying out various modifications or alterations under the spirit without departing from the utility model.

Please refer to Fig. 1 a to Figure 12.It should be noted that diagram provided in the present embodiment only illustrates in a schematic way The basic conception of the utility model, then in schema only display with related component in the utility model rather than when according to actual implementation Component count, shape and size draw, when actual implementation kenel, quantity and the ratio of each component can arbitrarily change for one kind Become, and its assembly layout kenel may also be increasingly complex.

Embodiment one

The utility model provides a kind of digital pcr system, please refers to Fig. 1 a to Fig. 1 g, and wherein Fig. 1 a is shown as the number The schematic perspective view of PCR system, Fig. 1 b are shown as the top view of the digital pcr system, and Fig. 1 c is shown as the number The bottom view of PCR system, Fig. 1 d, Fig. 1 e, Fig. 1 f and Fig. 1 g are respectively indicated as side of the digital pcr system on four direction View.Fig. 2 is shown as the decomposition texture schematic diagram of the digital pcr system, and in the present embodiment, the digital pcr system includes At least one droplet formation component 1 and drop spray orifice component 3, the drop spray orifice component 3 are connected to the droplet formation component 1 Lower section.

Specifically, the droplet formation component 1 includes at least one drop collecting tank 2.Show to be the liquid in Fig. 1 a The quantity that drop forms component 1 is one, and the droplet formation component 1 includes the situation of 4 drop collecting tanks 2, wherein each Drop collecting tank 2 is in line, and two neighboring drop collecting tank 2 shares a face side wall.Certainly, in other embodiments, more A drop collecting tank 2 can also use other arrangement modes, and discrete can be arranged, and the quantity of the droplet formation component 1 Or it is multiple, the protection scope of the utility model should not be excessively limited herein.

Referring to Fig. 3, being shown as the schematic perspective view of the drop spray orifice component 3, the drop spray orifice component 3 is wrapped Include several drop spray orifices 4.In the present embodiment, the drop spray orifice 4 is arranged in two rows, and every row drop spray orifice is uniformly distributed.? In other embodiments, the drop spray orifice 4 can also use other arrangement modes, should not excessively limit the utility model herein Protection scope.

Referring to Fig. 4, it is shown as the partial cutaway view of the drop spray orifice component 3, and in the present embodiment, the drop spray orifice 4 are open from the upper surface of the drop spray orifice component 3, and extend toward the lower surface direction of the drop spray orifice component 3, but do not pass through Wear the lower surface of the drop spray orifice component 3.The opening shape of the drop spray orifice 4 includes but is not limited to round, oval, more Any one in the shape of side.The volume of the drop spray orifice 4 determines the volume of digital pcr drop to be formed.

As an example, the drop spray orifice component 3 may include thermal printing chip.Thermal printing technique is printer field A major technique, the basic principle is that ink droplet sprayed by heating.In the utility model, the drop spray orifice component 3 Existing thermal printing chip can be used.

Please refer to Fig. 5 a to Fig. 5 h, wherein Fig. 5 a is shown as the positive stereochemical structure signal of the droplet formation component 1 Figure, Fig. 5 b are shown as the back side dimensional structure diagram of the droplet formation component 1, and Fig. 5 c is shown as the droplet formation component 1 top view, Fig. 5 d are shown as the bottom view of the droplet formation component 1, and Fig. 5 e, Fig. 5 f, Fig. 5 g and Fig. 5 h are respectively indicated as Side view of the droplet formation component 1 on four direction.In the present embodiment, 2 bottom of drop collecting tank is equipped with through slot 6, to expose multiple drop spray orifices 4, so that the drop spray orifice 4 is connected to the drop collecting tank 2.

Specifically, as shown in figure 4, vaporization member 5 is equipped in the drop spray orifice 4, for making in the drop spray orifice 4 In the vaporization of digital pcr aqueous liquids layer and the droplet formation oil being quickly pushed into the drop collecting tank 2, to form digital pcr Drop.

As an example, the vaporization member 5 is set to the bottom surface of the drop spray orifice 4, the vaporization member 5 can be used Heating element makes its vaporization by heating the digital pcr aqueous liquids layer.In the present embodiment, the heating element includes adding Backing, the heating sheet can be single metal layer, be also possible to composite multi-layer metal layer.The shape packet of the vaporization member 5 Circle or rectangular is included but is not limited to, area can be 0.5~2 times of 4 floor space of drop spray orifice.In other embodiments, The vaporization member 5 also can be set in the side wall of the drop spray orifice 4, should not excessively limit the protection of the utility model herein Range.

Specifically, the PCR system further includes at least one PCR reagent chamber for being used to store digital pcr solution, such as scheme Shown in 3 and Fig. 4, runner is equipped in the drop spray orifice component 3, the drop spray orifice 4 passes through the runner and the PCR reagent Chamber.

As an example, the runner includes at least one sprue 9 and a plurality of branch flow passage that connect with the sprue 9 10, each drop spray orifice 4 is connect with a branch flow passage 10 respectively.Show to be the drop spray orifice component 3 in Fig. 3 Situation including a sprue 9, in other embodiments, the quantity of the sprue 9 can also be with the drop collecting tank 2 Quantity match, such as shown in Figure 10 b be the drop spray orifice component 3 including four sprues 9 situation.

As an example, the material for constructing the runner and the drop spray orifice includes but is not limited to silicon, polymer, photoresist Deng.

Specifically, as shown in Figure 1a, the digital pcr system further includes pedestal 11, and the PCR reagent chamber is set to institute It states in pedestal 11, the drop spray orifice component 3 is connected to 11 top of pedestal.

As an example, the material of the pedestal 11 include but is not limited to transparent or opaque plastics, it is any in glass It is a kind of.

As an example, as shown in Fig. 2, the pedestal 11 include the first base assembly 12 and the second base assembly 13, it is described PCR reagent chamber includes PCR reagent upper chamber 14 and PCR reagent lower chambers 15, and the PCR reagent upper chamber 14 is from described first The upper surface of base assembly 12 is open, and runs through the lower surface of first base assembly 12, and the PCR reagent lower chambers 15 are certainly The upper surface of second base assembly 13 is open, and extends to the lower surface direction of second base assembly 13, but do not pass through The lower surface of second base assembly 13 is worn, the PCR reagent upper chamber 14 is connected to PCR reagent lower chambers 15 and part is handed over Repeatedly.

Please refer to Fig. 6 a to Fig. 6 g, wherein Fig. 6 a is shown as the schematic perspective view of the pedestal 11, and Fig. 6 b is shown as The top view of the pedestal 11, Fig. 6 c are shown as the bottom view of the pedestal 11, and Fig. 6 d, Fig. 6 e, Fig. 6 f and Fig. 6 g are shown respectively For side view of the pedestal 11 on four direction.

As an example, first base assembly 12 is fixed on described the by adhesive means such as double-sided adhesive or glue etc. Two base assemblies, 13 top.In the present embodiment, 13 surface of the second base assembly has one to be used to accommodate second pedestal The sink deck of component 13, and the quadrangle of the sink deck has arc-shaped wider space, the sink deck The protrusion of surrounding plays positioning action when first base assembly 12 pastes the sink deck surface.

Referring to Fig. 7, be shown as the partial top view of the pedestal, which show the PCR reagent upper chamber 14 with Relative positional relationship between the PCR reagent lower chambers 15, in the present embodiment, the PCR reagent be divided into two parts be in order to It is filled with enough digital pcr solution.

Fig. 8 and Fig. 9 is please referred to, schematic diagram of front three-dimensional structure and the back side of second base assembly 13 are respectively indicated as Schematic perspective view.As shown in Fig. 7 and Fig. 9, in the present embodiment, the lower surface of second base assembly 13 is equipped at least one A digital pcr solution injection hole 16, the digital pcr solution injection hole 16 is connected to the PCR reagent lower chambers 15, for leading to It crosses the digital pcr solution injection hole 16 and injects digital pcr solution in the PCR reagent chamber.

Specifically, as shown in Figures 7 and 8, the PCR reagent lower chambers 15 include first end and second end, the number PCR solution injection hole 16 is connected at the first end with the PCR reagent lower chambers 15, and the PCR reagent lower chambers 15 exist It is connected at the second end with the PCR reagent upper chamber 14.In the present embodiment, the size of the PCR reagent lower chambers 15 is certainly The first end to second extreme direction is gradually increased first, is then gradually reduced.It can prevent from adding using this design Bubble is formed when liquid.

Specifically, the lower surface of second base assembly 13 is equipped at least one gas vent 17 as shown in Fig. 7 and Fig. 9, The gas vent 17 is connected to by air flue 8 with the PCR reagent upper chamber 14.As shown in figure 8, the air flue 8 is from described second The upper surface of base assembly 13 is open, and extends to the lower surface direction of second base assembly 13, but through described the The lower surface of two base assemblies 13.

Specifically, the opening area of the digital pcr solution injection hole 16 is greater than the gas vent 17 in the present embodiment Opening area, the opening area of the digital pcr solution injection hole 16 do it is slightly bigger be rifle in order to support liquid-transfering gun Head.Due to capillary phenomenon, inject the indoor liquid of PCR reagent chamber will not from the digital pcr solution injection hole 16 or The gas vent 17 flows out.

Specifically, the digital pcr system further includes flexible circuit board 18 as shown in Fig. 1 a and Fig. 2, the soft route Plate 18 is connected to 11 top of pedestal, and the drop spray orifice component 3 is connected in the flexible circuit board 18.

Please refer to Figure 10 a to Figure 10 h, wherein Figure 10 a is shown as the drop spray orifice component 3 and is connected to the flexible wire The positive structure schematic of road plate 18, Figure 10 b are shown as the back that the drop spray orifice component 3 is connected to the flexible circuit board 18 Face structural schematic diagram, Figure 10 c are shown as the top view that the drop spray orifice component 3 is connected to the flexible circuit board 18, Figure 10 d It is shown as the bottom view that the drop spray orifice component 3 is connected to the flexible circuit board 18, Figure 10 e, Figure 10 f, Figure 10 g and figure 10h is respectively indicated as the drop spray orifice component 3 and is connected to side view of the flexible circuit board 18 on four direction.

Specifically, being equipped with the through-hole 19 for accommodating the drop spray orifice component 3 in the flexible circuit board 18, and described 19 surface of flexible circuit board is equipped with several first connections weld pad (not shown) and several second connection weld pads 20, the drop spray orifice Component 3 connect weld pad connection with described first by conducting wire, with by the flexible circuit board 18 by the vaporization member 5 and outside The connection of portion's controller.The drop spray orifice component 3 can be connect by routing (Wire Bond) technique of standard with described first Weld pad connection.

As an example, the flexible circuit board 18 is connect by adhesive means with the pedestal 11, it is as shown in Figure 6 a, described 11 surface of pedestal is equipped with for preventing glue from flowing to the channel 21 on the drop spray orifice component 3.The quantity of the channel 21 is It is multiple, it is distributed in the surrounding of the drop spray orifice component 3.The arrangement mode of the channel 21, which can according to need, to be adjusted, It is not limited to the mode that Fig. 6 a is presented.

Specifically, the droplet formation component 1 can also be fixed in the flexible circuit board 18 by adhesive means.Such as Shown in Fig. 5 b, 1 back side of droplet formation component is equipped with a boss 31, and this design can guarantee the droplet formation component 1 The strong bond between the flexible circuit board 18.

Specifically, being equipped at least two positional punches 22 in the flexible circuit board 18, such as shown in Figure 10 a and Figure 10 b Shown in Fig. 6 a, 11 surface of pedestal is equipped with positioning protrusion 23 corresponding with 22 position of positional punch, and the positioning is worn Hole 22 and the positioning protrusion 23 cooperate, and facilitate accurate positioning of the flexible circuit board 18 on the pedestal 11.

Specifically, the digital pcr system further includes a controller, Figure 11 is please referred to, is shown as the controller 24 Schematic perspective view, the controller 24 include controller housing 25 and the controller electricity in the controller housing 25 Road plate, the controller housing 25 have the supporting part 26 for placing the pedestal 11, if 26 surface of the supporting part is equipped with The dry circuit connection conductive pin 27 (also referred to as Pin) being connect with the controller circuitry connecting plate, the circuit connection conductive pin 27 is corresponding with the second connection position of weld pad 30 in the flexible circuit board 18.

Specifically, the controller 24 is connected with the drop spray orifice component 3 by flexible circuit board 18 through control institute The fever time, fever number and fever interval time of heating element are stated to control the formation speed of the digital pcr drop.Its In, the control circuit of the controller 24 can use existing circuit structure.

Specifically, as shown in figure 9, one end of the pedestal is provided at least one limiting slot 13, it is as shown in figure 11, described Controller housing 25 is provided at least one locating part 29 corresponding with the limiting slot 13.

Further, as can be seen from figures 8 and 9, the pedestal is additionally provided with a limiting through hole 7, and the limiting through hole 7 runs through The front and the back side of the pedestal, as shown in figure 11, the controller housing 25 are provided with corresponding with the limiting through hole 7 Locating part 30.

The digital pcr system of the utility model can be used for the formation of digital pcr drop, and the quick formation of drop depends on liquid Instant vaporization of the vaporization member in spray orifice to nanometer grade thickness liquid level is dripped, thus by the digital pcr solution in drop spray orifice Quickly to form digital pcr drop in push-in droplet formation oil, compared to the formation speed of 100 drops each second on the market, Droplet formation technology in the utility model may be implemented to be greater than 1000 droplet formation speed per second.Compared to oily Xiang Yushui The method that phase associated movement generates drop, the oil in the technical solution of the utility model is mutually static, therefore the consumption of oil phase Amount is greatly reduced, and reduces 50% or so oily phase dosage, has efficient digital pcr oil utilization rate.

Embodiment two

A kind of side being formed digital pcr drop using digital pcr system described in embodiment one is provided in the present embodiment Method, comprising the following steps: made in digital pcr solution boils and quickly push-in droplet formation oil using vaporization member, to form number Word PCR drop.

As an example, carrying out high-speed figure PCR droplet formation using thermal technology, the vaporization member includes heating part Part makes its vaporization by heating the digital pcr aqueous liquids layer.

Specifically, by the fever time of the control heating element, fever number and fever interval time to control State the formation speed of digital pcr drop.It can achieve using the digital pcr droplet formation method of the utility model greater than 1000 The digital pcr droplet formation speed of a/second.

As an example, the digital pcr droplet formation method the following steps are included:

S1: injecting digital pcr solution into PCR reagent chamber, enters digital pcr solution and the PCR reagent chamber The drop spray orifice of connection forms digital pcr aqueous liquids layer；

S2: droplet formation oil is added into drop collecting tank；

S3: it vaporizes the liquid level using vaporization member and is quickly pushed into the drop shape in the drop collecting tank At in oil, to form the digital pcr drop.

Specifically, the liquid level with a thickness of nanoscale, and be greater than 0.2nm, in the present embodiment, the thickness of the liquid level Degree ranges preferably from 0.2nm~30000nm.

Figure 12 is please referred to, the optics for being shown as the digital pcr drop formed using the digital pcr system of the utility model is aobvious Micro mirror figure, it is seen then that the digital pcr droplet morphology of formation is symmetrical, uniformly.

The digital pcr system of the utility model can satisfy the use of all digital pcr biochemical reagents.Due to many lifes The concentration of the significant molecule of object in blood is very low (such as Circulating tumor DNA only has 3 DNA moleculars in every 2ml blood), and The digital pcr system of the utility model has the characteristics that droplet formation quantity not by using oil mass to be limited and the characteristics of high speed, So that this kind of detection being applied in order to possible in digital pcr.

In conclusion the digital pcr system of the utility model carries out high-speed figure PCR droplet formation using thermal technology, Quick formed of drop vaporizes the transient heating of nanometer grade thickness liquid level dependent on the vaporization member in drop spray orifice, thus Digital pcr solution in drop spray orifice is quickly pushed into droplet formation oil to form digital pcr drop, it is every compared on the market The formation speed of second 100 drops, the droplet formation technology in the utility model may be implemented to be greater than 1000 liquid per second Drop forms speed.Compared to it is oily mutually and the water phase associated movement method that generates drop, oil in the technical solution of the utility model It is mutually static, therefore the consumption of oily phase is greatly reduced, and reduces 50% or so oily phase dosage, there is efficient number PCR oil utilization rate.So the utility model effectively overcomes various shortcoming in the prior art and has high industrial exploitation value Value.

The above embodiments are only illustrative of the principle and efficacy of the utility model, and not for limitation, this is practical new Type.Any person skilled in the art can all carry out above-described embodiment under the spirit and scope without prejudice to the utility model Modifications and changes.Therefore, such as those of ordinary skill in the art without departing from the revealed essence of the utility model All equivalent modifications or change completed under mind and technical idea, should be covered by the claim of the utility model.

Claims (21)

1. a kind of digital pcr system characterized by comprising

At least one droplet formation component, including at least one drop collecting tank；

Drop spray orifice component is connected to below the droplet formation component, including several drop spray orifices, and the drop spray orifice is from institute The upper surface opening of drop spray orifice component is stated, and is extended toward the lower surface direction of the drop spray orifice component, but not through described The lower surface of drop spray orifice component, the drop spray orifice are connected to the drop collecting tank, and vapour is equipped in the drop spray orifice Change component, for vaporizing the digital pcr aqueous liquids layer in the drop spray orifice and being quickly pushed into the drop collecting tank Droplet formation oil in, to form digital pcr drop.

2. digital pcr system according to claim 1, it is characterised in that: the drop spray orifice component includes thermal printing Chip.

3. digital pcr system according to claim 1, it is characterised in that: the vaporization member is set to the drop spray The bottom surface in hole or side wall.

4. digital pcr system according to claim 1, it is characterised in that: the opening shape of the drop spray orifice includes circle Shape, ellipse, any one in polygon.

5. digital pcr system according to claim 1, it is characterised in that: the vaporization member includes heating element, is passed through Heating the digital pcr aqueous liquids layer makes its vaporization.

6. digital pcr system according to claim 5, it is characterised in that: the heating element includes at least one layer of metal Layer.

7. digital pcr system according to claim 1, it is characterised in that: the drop collects trench bottom and is equipped with through slot, institute It states through slot and exposes multiple drop spray orifices.

8. digital pcr system according to claim 1, it is characterised in that: the PCR system further includes that at least one is used for The PCR reagent chamber of digital pcr solution is stored, runner is equipped in the drop spray orifice component, the drop spray orifice passes through described Runner and the PCR reagent chamber.

9. digital pcr system according to claim 8, it is characterised in that: the runner include at least one sprue and The a plurality of branch flow passage connecting with the sprue, each drop spray orifice are connect with a branch flow passage respectively.

10. digital pcr system according to claim 8, it is characterised in that: the digital pcr system further includes pedestal, institute It states PCR reagent chamber to be set in the pedestal, the drop spray orifice component is connected to above the pedestal.

11. digital pcr system according to claim 10, it is characterised in that: the pedestal include the first base assembly with Second base assembly, the PCR reagent chamber include PCR reagent upper chamber and PCR reagent lower chambers, the PCR reagent epicoele Room is open from the upper surface of first base assembly, and runs through the lower surface of first base assembly, under the PCR reagent Chamber is open from the upper surface of second base assembly, and extends to the lower surface direction of second base assembly, but not Through the lower surface of second base assembly, the PCR reagent upper chamber is connected to PCR reagent lower chambers and part is folded.

12. digital pcr system according to claim 11, it is characterised in that: the lower surface of second base assembly is set There is at least one digital pcr solution injection hole, the digital pcr solution injection hole is connected to the PCR reagent lower chambers.

13. digital pcr system according to claim 11, it is characterised in that: the PCR reagent lower chambers include first end And second end, the digital pcr solution injection hole are connected at the first end with the PCR reagent lower chambers, the PCR examination Agent lower chambers are connected at the second end with the PCR reagent upper chamber, and the sizes of the PCR reagent lower chambers is from described One end to second extreme direction is gradually increased first, is then gradually reduced.

14. digital pcr system according to claim 11, it is characterised in that: the lower surface of second base assembly is set There is at least one gas vent, the gas vent is connected to by air flue with the PCR reagent upper chamber, and the air flue is from described second The upper surface of base assembly is open, and extends to the lower surface direction of second base assembly, but does not run through second base The lower surface of holder assembly.

15. digital pcr system according to claim 11, it is characterised in that: first base assembly passes through gluing side Formula is fixed on above second base assembly.

16. digital pcr system according to claim 10, it is characterised in that: the digital pcr system further includes flexible wire Road plate, the flexible circuit board are connected to above the pedestal, are equipped in the flexible circuit board for accommodating the drop spray The through-hole of aperture member, and the flexible circuit board surface is equipped with several first connection weld pads and several second connection weld pads, it is described Drop spray orifice component connect weld pad connection with described first by conducting wire.

17. digital pcr system according to claim 16, it is characterised in that: the flexible circuit board passes through adhesive means It is connect with the pedestal, the base-plates surface is equipped with for preventing glue from flowing to the channel on the drop spray orifice component.

18. digital pcr system according to claim 16, it is characterised in that: be equipped at least two in the flexible circuit board A positional punch, the base-plates surface are equipped with positioning protrusion corresponding with the positional punch position.

19. digital pcr system according to claim 16, it is characterised in that: the digital pcr system further includes a control Device, the controller include controller housing and the controller circuit board in the controller housing, outside the controller Shell has the supporting part for placing the pedestal, and the supporting part surface is equipped with several and controller circuitry connecting plate and connects The circuit connection conductive pin connect, the circuit connection conductive pin are corresponding with the second connection position of weld pad.

20. digital pcr system according to claim 19, it is characterised in that: one end of the pedestal is provided at least one A limiting slot, the controller housing are provided at least one locating part corresponding with the limiting slot.

21. digital pcr system according to claim 19, it is characterised in that: the pedestal is provided with a limiting through hole, institute Front and the back side that limiting through hole runs through the pedestal are stated, the controller housing is provided with corresponding with the limiting through hole Locating part.