CN106635785B - A kind of PCR fluorescence detector - Google Patents
A kind of PCR fluorescence detector Download PDFInfo
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- CN106635785B CN106635785B CN201611245178.8A CN201611245178A CN106635785B CN 106635785 B CN106635785 B CN 106635785B CN 201611245178 A CN201611245178 A CN 201611245178A CN 106635785 B CN106635785 B CN 106635785B
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
- B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
- B01L2300/1827—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks using resistive heater
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0442—Moving fluids with specific forces or mechanical means specific forces thermal energy, e.g. vaporisation, bubble jet
- B01L2400/0451—Thermophoresis; Thermodiffusion; Soret-effect
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- Dispersion Chemistry (AREA)
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- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The present invention provides a kind of PCR fluorescence detector, in the detector, PCR reaction tube is placed in first through hole, the second through-hole and third through-hole, it is located at PCR reaction tube bottom end at third through-hole, the liquid level of PCR reaction solution is located at first through hole in PCR reaction tube, realize that 95 DEG C of heating plates heat PCR reaction tube bottom end, 60 DEG C of heating plates are heated at PCR reaction solution liquid level in PCR reaction tube, and then the upper and lower side of PCR reaction tube generates the temperature difference.The temperature difference makes the PCR reaction solution in PCR reaction tube flow to top low temperature region by bottom high-temperature area, forms convection current.Due to the temperature difference and convection current, DNA can constantly be expanded in PCR fluorescence detector, do not need as temperature needed for reaching DNA cloning different phase and continuous heating, cooling, therefore, PCR fluorescence detector provided by the invention can save proliferation time, and then shorten the detection time of PCR detector.
Description
Technical field
The present invention relates to detection device technology field more particularly to a kind of PCR fluorescence detectors.
Background technique
PCR (Polymerase Chain Reaction, polymerase chain reaction) technology is a kind of in vitro amplification
Expand the Protocols in Molecular Biology of specific DNA (deoxyribonucleic acid, DNA) sequence.Round pcr tool
Have the characteristics that high specificity, high sensitivity, purity requirement are low and easy, quick, thus is widely used in molecular biology
Detection and analysis.
In general, the process of round pcr DNA amplification are as follows: in suitable buffer solution and the mixture of hot resistant DNA polymerase
In system, DNA forms 2 complementations in about 95 DEG C of at a temperature of generation high temperature solution chain reaction, i.e. the hydrogen bond fracture of DNA double interchain
Single stranded DNA；In the case where the oligonucleotide chain with directionality and specificity is mediated as primer annealing (renaturation) occurs for single stranded DNA
The temperature of single stranded DNA, i.e., be rapidly decreased in the range of the design temperature value (typically about 50-65 DEG C) of primer by reaction, single
In conjunction with chain DNA with primer follows base pair complementarity principle；Temperature is increased to rapidly 72 DEG C or so again, single stranded DNA and primer
In conjunction with rear generation extension, archaeal dna polymerase combines deoxynucleoside triphosphate holding since primer 3 ', according to corresponding in template
Base is sequentially extended with complementary pairing principle, to form a new DNA fragmentation complementary with template.Expand by round pcr
After increasing, initial DNA molecular quantity is doubled, and is recycled for one；DNA molecular after multiplication then becomes next circulation
Template is so gone down, and after 30-40 circulation, DNA molecular number will be amplified to nearly the 10 of initial value9Times.
Currently, existing PCR detector can simultaneously expand the PCR reaction solution in multiple PCR reaction tubes.It is examined in PCR
When surveying instrument DNA amplification, the PCR reaction tube for having contained PCR reaction solution is placed in sample well, tungsten halogen lamp is as radiation source.Halogen tungsten
The light that lamp issues after five color light source optical filters by being irradiated on each PCR reaction tube, the fluorescence in each PCR reaction solution point
Son can be generated fluorescence by the excitation of tungsten halogen lamp light, and fluorescence reaches CCD (Charge- after passing through multicolored fluorescent optical filter
Coupled Device, charge coupled cell) camera, fluorescence signal is converted into electric signal by CCD camera, and then judge PCR
Product total amount after DNA amplification.PCR since the distance of each PCR reaction tube to tungsten halogen lamp is different, closer to tungsten halogen lamp center
Reaction tube, internal fluorescent molecule is easier to be excited, and fluorescence is stronger, and the PCR far from tungsten halogen lamp center reacts
The fluorescence of pipe, internal fluorescent molecule is weaker, thus has differences with the fluorescence detection after a pcr amplified DNA.Separately
Outside, it during PCR detector DNA amplification, needs constantly to be brought rapidly up, cool down, and the time of heating, cooling is longer, this is big
Extend the time of an amplification cycles greatly, and then reduces amplification efficiency.
Summary of the invention
The present invention provides a kind of PCR fluorescence detector, to solve the problems, such as that the existing PCR detector test time is longer.
The present invention provides a kind of PCR fluorescence detector, and the detector includes circuit device and is arranged in the circuit
Light path device on device upper surface；
The inside of the light path device has hollow cavity, and the top of the light path device has and the hollow cavity phase
The first through hole of connection；
Be provided with 60 DEG C of heating plates at the top of the light path device, 60 DEG C of heating plates to be provided centrally with second logical
Hole；
95 DEG C of heating plates are provided in the hollow cavity, 95 DEG C of heating plates are provided centrally with third through-hole；
The center of the first through hole, second through-hole and the third through-hole is on the same line.
Preferably, the circuit device includes detection circuit board, optical transmitting set and optical receiver, the optical transmitting set and institute
It states optical receiver to be arranged in the detection circuit board, and the line between the optical transmitting set and the detection circuit board center
Perpendicular to the line between the optical receiver and the detection circuit board center.
Preferably, be arranged four groups in the detection circuit board described in optical transmitting set and the optical receiver, light described in four groups
The rounded distribution in the detection circuit board of transmitter and the optical receiver；Optical transmitting set described in every two groups is adjacent, and
It is oppositely arranged with optical transmitting set described in remaining two groups.
Preferably, the optical transmitting set is light emitting diode, and the optical receiver is photodiode.
Preferably, the light path device includes hollow first shell and second shell, wherein
The first shell includes being parallel to the first upper shell of the detection circuit board and perpendicular to the detection circuit
The first lower house of plate, the first upper shell and the first lower house are connected by inclined first reflective mirror；
The second shell includes being parallel to the second upper shell of the detection circuit board and perpendicular to the detection circuit
The second lower house of plate, the second upper shell and the second lower house are connected by inclined second reflective mirror；
The first upper shell and the second upper shell are mutually perpendicular to, and the first upper shell and second upper casing
The end of body is not attached to.
Preferably, the first filter is arranged in the inside of the first lower house and the first lens, first lens are located at institute
It states between the first filter and the optical transmitting set；
The second filter and the second lens are arranged in the inside of the second lower house, and second lens are located at second filter
Between mirror and the optical receiver.
Preferably, the light path device includes first shell and the second shell described in four groups, first shell described in four groups
Body and the second shell are circumferentially uniformly distributed around the first through hole.
Preferably, 60 DEG C of heating plates include the first lower plate from bottom to top set gradually, in first resistor and first
Plate；95 DEG C of heating plates include the second lower plate, second resistance and the second upper plate from bottom to top set gradually.
Preferably, detector further includes light path device pedestal, the internal structure and the optical path of the light path device pedestal
The external structure of device matches.
Preferably, the detector further includes heat sink and radiator fan, and the heat sink setting is heated at described 60 DEG C
The outer surface of the light path device pedestal, and the radiator fan and described 95 DEG C are arranged in the top of plate, the radiator fan
Heating plate is located in same level.
The technical solution that the embodiment of the present invention provides can include the following benefits:
In PCR fluorescence detector provided by the invention, PCR reaction tube is placed in first through hole, the second through-hole and third
In through-hole, and it is located at the bottom end of PCR reaction tube at third through-hole, it is logical to be located at first for the liquid level of PCR reaction solution in PCR reaction tube
At hole, to realize that 95 DEG C of heating plates heat the bottom end of PCR reaction tube, 60 DEG C of heating plates are to PCR reaction solution in PCR reaction tube
It is heated at liquid level, so that the upper and lower side of PCR reaction tube generates the temperature difference.Since the presence of the temperature difference makes in PCR reaction tube
PCR reaction solution forms convection current by the low-temperature region at the top of the high-temperature area flow direction of bottom.Due to existing in PCR reaction tube
The convection current of the temperature difference and PCR reaction solution, thus DNA can constantly be expanded in PCR fluorescence detector, not needed to reach DNA
Temperature and the continuous heating, cooling in PCR detector needed for expanding different phase, therefore, PCR fluorescence inspection provided by the invention
Proliferation time can be saved by surveying instrument, and then shorten the detection time of PCR detector.
It should be understood that above general description and following detailed description be only it is exemplary and explanatory, not
It can the limitation present invention.
Detailed description of the invention
The drawings herein are incorporated into the specification and forms part of this specification, and shows and meets implementation of the invention
Example, and be used to explain the principle of the present invention together with specification.
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, for those of ordinary skill in the art
Speech, without any creative labor, is also possible to obtain other drawings based on these drawings.
Fig. 1 is the one package structual schematic diagram of PCR fluorescence detector provided in an embodiment of the present invention；
Fig. 2 is the assembly structure diagram of PCR fluorescence detector provided in an embodiment of the present invention；
Fig. 3 is the location map of PCR reaction tube provided in an embodiment of the present invention；
Fig. 4 is the structural schematic diagram of 60 DEG C of heating plates provided in an embodiment of the present invention；
Fig. 5 is the structural schematic diagram of 95 DEG C of heating plates provided in an embodiment of the present invention；
Fig. 6 is four groups of path channels schematic diagrames provided in an embodiment of the present invention；
Symbol indicates:
1- circuit device, 2- light path device, 3- hollow cavity, 4- first through hole, 5-60 DEG C of heating plate, the second through-hole of 6-,
7-95 DEG C of heating plate, 8- third through-hole, 9- detection circuit board, 10- optical transmitting set, 11- optical receiver, 12- first shell, 13-
Second shell, 14- first upper shell, 15- first lower house, the first reflective mirror of 16-, 17- second upper shell, the second lower casing of 18-
Body, the second reflective mirror of 19-, the first filter of 20-, the first lens of 21-, the second filter of 22-, the second lens of 23-, the first lower plate of 24-,
25- first resistor, the first upper plate of 26-, the second lower plate of 27-, 28- second resistance, the second upper plate of 29-, 30- light path device pedestal,
31- heat sink, 32- radiator fan, 33-PCR reaction tube.
Specific embodiment
Example embodiments are described in detail here, and the example is illustrated in the accompanying drawings.Following description is related to
When attached drawing, unless otherwise indicated, the same numbers in different drawings indicate the same or similar elements.Following exemplary embodiment
Described in embodiment do not represent all embodiments consistented with the present invention.On the contrary, they be only with it is such as appended
The example of device and method being described in detail in claims, some aspects of the invention are consistent.
Attached drawing 1 and attached drawing 2 are please referred to, attached drawing 1 shows the packing knot of PCR fluorescence detector provided in an embodiment of the present invention
Structure schematic diagram, attached drawing 2 show the assembly structure diagram of PCR fluorescence detector provided in an embodiment of the present invention.
PCR fluorescence detector provided in an embodiment of the present invention includes circuit device 1 and light path device 2, and light path device 2 is arranged
On 1 upper surface of circuit device, wherein circuit device 1 is used to issue the light of PCR fluorescence detection and receives fluorescence, Jin Erfen
Product total amount after analysing pcr amplified DNA；Light path device 2 provides optical path for providing the track route of light and fluorescence.
Specifically, circuit device 1 includes detection circuit board 9, optical transmitting set 10 and optical receiver 11, and detection circuit board 9 is
Substrate, optical transmitting set 10 and optical receiver 11 are arranged on the upper surface of detection circuit board 9.Optical transmitting set 10 shines for issuing
The light being mapped on PCR reaction tube 33, the fluorescent material in PCR reaction tube 33 are generated by fluorescence is generated after the excitation of light
Fluorescence received by optical receiver 11, received fluorescence conversion is electric signal by optical receiver 11, and then after analyzing DNA cloning
Product total amount.
Further, the line between 9 center of optical transmitting set 10 and detection circuit board is perpendicular to optical receiver 11 and detection electricity
Line between 9 center of road plate, so that the fluorescence that the light excitation fluorescent molecule that optical transmitting set 10 issues generates can be connect by light
It receives device 11 to receive, and the difference of fluorescence signal power is not present.
In embodiments of the present invention, light path device 2 includes first shell 12 and second shell 13, first shell 12 and second
Shell 13 be it is hollow, in order to inside be arranged eyeglass.Specifically, first shell 12 includes being parallel to detection circuit board 9
First upper shell 14 and first lower house 15 perpendicular to detection circuit board 9, first upper shell 14 and first lower house 15 are logical
The first reflective mirror 16 is crossed to be connected；Second shell 13 include be parallel to detection circuit board 9 second upper shell 17 and perpendicular to
The second lower house 18 of detection circuit board 9, second upper shell 17 and second lower house 18 are connected by the second reflective mirror 19.The
The light that optical transmitting set 10 issues can be reflected on PCR reaction tube 33 by one reflective mirror 16 and the second reflective mirror 19, and then be made
It obtains the fluorescent molecule in PCR reaction tube 33 and receives the strong and weak identical exciting light of illumination, generate the identical fluorescence of fluorescence intensity, avoid
PCR detection is influenced because fluorescence is strong and weak different.
The setting of the position in detection circuit board 9, first shell 12 and second according to optical transmitting set 10 and optical receiver 11
Shell 13 is also mutually perpendicular to, i.e. first upper shell 14 and second upper shell 17 is mutually perpendicular to, to cooperate optical transmitting set 10 and detection
Circuit board 9 forms complete sending optical path and receiving light path.Since the top of light path device 2 has first through hole 4, thus, the
The end of one upper housing 14 and second upper shell 17 is mutually perpendicular to, and is not attached to.
Further, the first filter 20 and the first lens 21 are arranged in the inside of first lower house 15, and the first lens 21 are located at
Between first filter 20 and optical transmitting set 10.Inside the second filter 22 of setting of second lower house 18 and the second lens 23, and the
Two lens 23 are between the second filter 22 and optical receiver 11.First filter 20 and the second filter 22 are respectively used to filter out light hair
The illumination that emitter 10 and fluorescent molecule are issued, and then obtain using required lighting color, such as feux rouges, green light or yellow light
Deng.First lens 21 are used to that the first reflective mirror 16 will to be irradiated to after illumination that optical transmitting set 10 issues cohesion, convenient for generate compared with
Strong illumination.Second lens 23 are used to agglomerate the light beam dispersed by the second reflective mirror 19 for a branch of stronger illumination, in turn
Fluorescence signal is converted into electric signal convenient for optical receiver 11.
In PCR fluorescence detector provided in an embodiment of the present invention, the inside of light path device 2 has hollow cavity 3, hollow
Inner cavity 3 is for placing 95 DEG C of heating plates 7, so that bottom-heated of 95 DEG C of heating plates 7 to PCR reaction tube 33, in PCR reaction tube 33
The PCR reaction solution of bottom has 95 DEG C of temperature.The top of light path device 2 is provided with 60 DEG C of heating plates 5, so that 60 DEG C of heating plates
It is heated at the nozzle of 5 pairs of PCR reaction tubes 33, the temperature in PCR reaction tube 33 at PCR reaction solution liquid level with 60 DEG C.
The top of light path device 2 has the center setting of 4, the 60 DEG C of heating plates 5 of first through hole being connected with hollow cavity 3
There is the second through-hole 6, likewise, 95 DEG C of heating plates 7 are provided centrally with third through-hole 8.First through hole 4, the second through-hole 6 and
Three through-holes 8 are used to place PCR reaction tube 33, and 95 DEG C of heating plates 7 are located at the bottom of PCR reaction tube 33, and 60 DEG C heating plate 5
In the nozzle position of PCR reaction tube 33, attached drawing 3 is specifically please referred to.For the placement convenient for PCR reaction tube 33, first through hole 4,
The center of two through-holes 6 and third through-hole 8 is on the same line.
Since the temperature at 33 nozzle of PCR reaction tube is 60 DEG C, the temperature of bottom is 95 DEG C, thus PCR reaction solution is close
Temperature gradient is presented on gravity direction in the PCR reaction tube 33 closed.Since the presence of temperature gradient makes upper layer and lower layer PCR anti-
It answers the density of liquid different, then forms density gradient.Density gradient can create antagonism the floating of gravity, internal chamber wall and internal friction
Power, so that the high temperature PCR reaction solution of 33 bottom of PCR reaction tube is flowed to the low temperature PCR reaction solution at top, the low temperature at top
PCR reaction solution is flowed to the high temperature PCR reaction solution of bottom, forms a circulation.Due to the heating of the 95 DEG C of heating plates 7 in bottom, make
It obtains above-mentioned circulation constantly to carry out, PCR reaction solution also constantly undergoes high temperature, low temperature, so that it is special to complete the DNA in PCR reaction solution
Determine the amplification at temperature, the amplification under specific temperature is reached without frequently changing heating temperature, it is anti-that this can accelerate PCR
Speed is answered, PCR detection time is shortened.
Attached drawing 4 and attached drawing 5 are please referred to, attached drawing 4 and attached drawing 5 respectively illustrate the knot of 60 DEG C of heating plate 5 and 95 DEG C heating plates 7
Structure schematic diagram.
60 DEG C of heating plates 5 include the first lower plate 24, first resistor 25 and the first upper plate 26 from bottom to top set gradually,
In, the first lower plate 24 and the first upper plate 26 are fixed plate, for fixing first resistor 25.The use of first resistor 25 is buried resistance mode and is set
It sets between the first lower plate 24 and the first upper plate 26, to play the role of heating.First resistor 25 is uniform around the second through-hole 6
Setting, and then the PCR reaction solution in PCR reaction tube 33 is heated by the second through-hole 6.The quantity of first resistor 25 can be according to electricity
The size of resistance is specifically arranged.
95 DEG C of heating plates 7 include the second lower plate 27, second resistance 28 and the second upper plate 29 from bottom to top set gradually,
In, the second lower plate 27 and the second upper plate 29 are fixed plate, for fixing second resistance 28.The use of second resistance 28 is buried resistance mode and is set
It sets between the second lower plate 27 and the second upper plate 29, to play the role of heating.Second resistance 28 is uniform around third through-hole 8
Setting, and then the PCR reaction solution in PCR reaction tube 33 is heated by third through-hole 8.Likewise, the quantity of second resistance 28 can
To be specifically arranged according to the size of resistance.
PCR fluorescence detector provided in an embodiment of the present invention further includes light path device pedestal 30, and light path device pedestal 30 is set
It sets in the outer surface of light path device 2, and then plays the role of Backup lightpath device 2.For can be compared with convenient for light path device pedestal 30
The internal structure of Backup lightpath device 2 well, light path device pedestal 30 matches with the external structure of light path device 2.
Since the top of light path device 2 is provided with 60 DEG C of heating plates 5,19 distance 60 of the first reflective mirror 16 and the second reflective mirror
DEG C heating plate 5 is relatively close；And light path device 2 is internally provided with 95 DEG C of heating plates 7, the first filter 20, the filter of the first lens 21, second
Mirror 22 and 23 95 DEG C of heating plates 7 of equal distance of the second lens are relatively close, thus after long-time use, the first reflective mirror 16, the
The eyeglasses such as two reflective mirrors 19 and the first filter 20 easily deform under the action of high temperature, and the serious shadow of deformed eyeglass
Ring PCR fluorescence detection.To prevent the eyeglasses such as the first reflective mirror 16, the second reflective mirror 19 and the first filter 20 from deforming, this
The PCR fluorescence detector that inventive embodiments provide further includes heat sink 31 and radiator fan 32.The setting of heat sink 31 adds at 60 DEG C
The top of hot plate 5 to take away the heat near the first reflective mirror 16 and the second reflective mirror 19, while being taken away 95 DEG C of heating plates 7 and being produced
Raw partial heat.The outer surface of light path device pedestal 30, and radiator fan 32 and 95 DEG C heating plate 7 is arranged in radiator fan 32
In same level, to take away the heat of 95 DEG C of heating plates 7 generation, and then the eyeglasses such as the first reflective mirror 16 is avoided to become
Shape.
The detection process of PCR fluorescence detector provided in an embodiment of the present invention are as follows: the PCR reaction solution mixed will be loaded with
PCR reaction tube 33 be placed in first through hole 4, the second through-hole 6 and third through-hole 8, start 60 DEG C of heating plates 5 and 95 DEG C plus
Hot plate 7 heats, and to form fluid convection, DNA is constantly expanded.Meanwhile during DNA is constantly expanded, optical transmitting set 10 is sent out
Light beam is radiated on the first lens 21 out, and the first lens 21 pass through the first filter 20 after agglomerating light beam, to obtain required light
According to color.Determine that the light beam of lighting color is radiated on the first reflective mirror 16, the first reflective mirror 16 shines light beam after 90 ° of reflections
It penetrates on PCR reaction tube 33, the fluorescent molecule in PCR reaction tube 33 receives the strong and weak identical exciting light of illumination, and then generates glimmering
The identical fluorescence of luminous intensity.The fluorescence that 33 different direction of PCR reaction tube issues is radiated on the second reflective mirror 19, the second reflective mirror
19 will be radiated on the second filter 22 after 90 ° of fluorescence reflections, to filter out remaining photochromic light beam.The light to filter through the second filter 22
Beam is radiated on the second lens 23, is received after 23 optically focused of the second lens by optical receiver 11, and then fluorescence signal is converted to electricity
Signal completes PCR fluorescence detection.
In above-mentioned detection process, fluorescent molecule can receive the strong and weak identical exciting light of illumination, and then it is strong to generate fluorescence
Identical fluorescence is spent, avoids influencing PCR detection because fluorescence is strong and weak different.
In PCR fluorescence detector provided in an embodiment of the present invention, four groups of 10 Hes of optical transmitting set are set in detection circuit board 9
Optical receiver 11 is issued with forming four groups of light, receives combination.Correspondingly, light path device 2 includes four groups of first shells 12 and second
It is logical that shell 13, four groups of optical transmitting sets 10 and optical receiver 11 and four groups of first shells 12 and second shell 13 form four groups of optical paths
Road specifically please refers to attached drawing 6, wherein 1 to 4 is optical transmitting set 10, and 1-1 to 4-4 is optical receiver 11.
The rounded distribution in detection circuit board 9 of four groups of optical transmitting sets 10 and optical receiver 11, every two groups of optical transmitting sets 10
It is disposed adjacent, and is oppositely arranged with remaining two groups of optical transmitting set 10, if 1 in attached drawing 6 is disposed adjacent with 2,3 are disposed adjacent with 4,
But it 1,2 is oppositely arranged with 3,4.Since the line between 9 center of optical transmitting set 10 and detection circuit board is perpendicular to optical receiver 11
Line between 9 center of detection circuit board, thus every two groups of optical receivers 11 are disposed adjacent, and with remaining two groups of optical receiver
11 are oppositely arranged, and if the 1-1 in attached drawing 6 is disposed adjacent with 2-2,3-3 is disposed adjacent with 4-4, but 1-1,2-2 and 3-3,4-4 phase
To setting.Certainly, optical transmitting set 10 and optical receiver 11 are not limited to four groups, first shell 12 and second shell 13 also not office
It is limited to four groups, the setting of other groups of numbers is also in the range of the application protects.
During PCR fluorescence detector provided in an embodiment of the present invention amplification, detection DNA, put in PCR reaction tube 33
Four groups of different DNA moleculars are set, mark different fluorescent molecules respectively for four groups of different DNA moleculars；Four groups of path channels
The light beam of different optical path and excitation fluorescent molecule fluorescence can be provided respectively, and then PCR provided in an embodiment of the present invention is glimmering
Optical detector can detect four kinds of different DNA moleculars simultaneously.When detecting four groups of different DNA moleculars, four groups of light hairs are controlled
Emitter 10 and optical receiver 11 unlatch and close the time, make unlatching and the pass of two adjacent groups optical transmitting set 10 and optical receiver 11
It closes between the time that there are the time differences, and then receives corresponding fluorescence convenient for different optical receivers 11, PCR fluorescence detector is avoided to go out
Existing detection error.
In the various embodiments described above provided by the invention, optical transmitting set 10 is preferably light emitting diode, and optical receiver 11 is excellent
It is selected as photodiode.
Those skilled in the art will readily occur to of the invention its after considering specification and the disclosure invented here of practice
Its embodiment.This application is intended to cover any variations, uses, or adaptations of the invention, these modifications, purposes or
Person's adaptive change follows general principle of the invention and including the undocumented common knowledge in the art of the present invention
Or conventional techniques.The description and examples are only to be considered as illustrative, and true scope and spirit of the invention are by following
Claim is pointed out.
It should be understood that the relational terms of such as " first " and " second " or the like be used merely to an entity or
Operation is distinguished with another entity or operation, and without necessarily requiring or implying between these entities or operation, there are any
This actual relationship or sequence.The present invention is not limited to the precise structure already described above and shown in the accompanying drawings,
And various modifications and changes may be made without departing from the scope thereof.The scope of the present invention is only limited by the attached claims
System.
Claims (7)
1. a kind of PCR fluorescence detector, which is characterized in that the detector includes circuit device (1) and is arranged in the electricity
Light path device (2) on road device (1) upper surface；
The circuit device (1) includes the four groups of light emittings of detection circuit board (9) and setting on the detection circuit board (9)
Device (10) and optical receiver (11), and the line between the optical transmitting set (10) and the detection circuit board (9) center is vertical
Line between the optical receiver (11) and the detection circuit board (9) center；
The inside of the light path device (2) has hollow cavity (3), the top of the light path device (2) have with it is described hollow
The first through hole (4) that inner cavity (3) is connected；
60 DEG C of heating plates (5) are provided at the top of the light path device (2), 60 DEG C of heating plates (5) are provided centrally with
Two through-holes (6)；
95 DEG C of heating plates (7) are provided in the hollow cavity (3), the third that is provided centrally with of 95 DEG C of heating plates (7) is led to
Hole (8)；
The center of the first through hole (4), second through-hole (6) and the third through-hole (8) is on the same line；
The light path device (2) includes four groups of hollow first shells (12) and second shell (13), wherein
The first shell (12) includes being parallel to the first upper shell (14) of the detection circuit board (9) and perpendicular to the inspection
The first lower house (15) of slowdown monitoring circuit plate (9), the first upper shell (14) and the first lower house (15) pass through inclined
First reflective mirror (16) is connected；
The second shell (13) includes being parallel to the second upper shell (17) of the detection circuit board (9) and perpendicular to the inspection
The second lower house (18) of slowdown monitoring circuit plate (9), the second upper shell (17) and the second lower house (18) pass through inclined
Second reflective mirror (19) is connected；
The first upper shell (14) and the second upper shell (17) are mutually perpendicular to, and the first upper shell (14) and described
The end of second upper shell (17) is not attached to；
The inside setting the first filter (20) of the first lower house (15) and the first lens (21), the first lens (21) position
Between first filter (20) and the optical transmitting set (10)；
The inside setting the second filter (22) of the second lower house (18) and the second lens (23), the second lens (23) position
Between second filter (22) and the optical receiver (11).
2. PCR fluorescence detector according to claim 1, which is characterized in that be arranged four groups on the detection circuit board (9)
The optical transmitting set (10) and the optical receiver (11), optical transmitting set described in four groups (10) and the optical receiver (11) are in institute
State rounded distribution on detection circuit board (9)；Optical transmitting set (10) described in every two groups is adjacent, and sends out with light described in remaining two groups
Emitter (10) is oppositely arranged.
3. PCR fluorescence detector according to claim 1, which is characterized in that the optical transmitting set (10) is light-emitting diodes
Pipe, the optical receiver (11) are photodiode.
4. PCR fluorescence detector according to claim 1, which is characterized in that the light path device (2) includes described in four groups
First shell (12) and the second shell (13), first shell described in four groups (12) and the second shell (13) are around described
First through hole (4) is circumferentially uniformly distributed.
5. PCR fluorescence detector according to claim 1, which is characterized in that 60 DEG C of heating plates (5) include by lower and
On the first lower plate (24), first resistor (25) and the first upper plate (26) that set gradually；95 DEG C of heating plates (7) include under
The second lower plate (27), second resistance (28) and the second upper plate (29) set gradually on and.
6. according to claim 1, PCR fluorescence detector described in any one of 2,4,5, which is characterized in that detector further includes
Light path device pedestal (30), the internal structure and the external structure phase of the light path device (2) of the light path device pedestal (30)
It coincide.
7. PCR fluorescence detector according to claim 6, which is characterized in that the detector further includes heat sink (31)
With radiator fan (32), the heat sink (31) is arranged at the top of 60 DEG C of heating plates (5), and the radiator fan (32) sets
It sets in the outer surface of the light path device pedestal (30), and the radiator fan (32) and 95 DEG C of heating plates (7) are located at together
On one horizontal plane.
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| WO2018119848A1 (en) * | 2016-12-29 | 2018-07-05 | 湖南圣湘生物科技有限公司 | Pcr fluorescence detector |
| CN109554295B (en) * | 2019-01-21 | 2022-03-29 | 武汉理工大学 | PCR amplification and disease detection device for ocean-going crew |
| CN110272822B (en) * | 2019-06-06 | 2021-10-26 | 上海交通大学 | Gene amplification real-time fluorescence quantitative detection device and detection method |
| CN111304051B (en) * | 2020-02-20 | 2023-08-11 | 珠海黑马生物科技有限公司 | PCR instrument and use method thereof |
| CN111647504B (en) * | 2020-06-10 | 2023-07-04 | 赵毅 | Quick PCR reaction tube and instrument thereof |
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Address after: 410205 Changsha province high and New Technology Industrial Development Zone, Lu Pine Road, No. 680, Hunan Patentee after: Shengxiang Biotechnology Co., Ltd Address before: 410205 Changsha province high and New Technology Industrial Development Zone, Lu Pine Road, No. 680, Hunan Patentee before: Sansure Biotech Inc. |