CN107937268A - Portable constant temperature nucleic acid augmentative instrument - Google Patents
Portable constant temperature nucleic acid augmentative instrument Download PDFInfo
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- CN107937268A CN107937268A CN201711182767.0A CN201711182767A CN107937268A CN 107937268 A CN107937268 A CN 107937268A CN 201711182767 A CN201711182767 A CN 201711182767A CN 107937268 A CN107937268 A CN 107937268A
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- sample
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- amplified reaction
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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
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- Life Sciences & Earth Sciences (AREA)
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- General Health & Medical Sciences (AREA)
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- Chemical Kinetics & Catalysis (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
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Abstract
The present invention provides a kind of portable constant temperature nucleic acid augmentative instrument, including：One sample process module, to carry out sample process to genetic test sample, and obtains sample treatment solution；One amplified reaction module, to make sample treatment solution carry out amplified reaction in a reaction temperature section；One control module, built-in control program are described to control program to control sample process module and amplified reaction module to perform an operation for determining step.Its external form is light, can carry, it can be achieved that key start is without other parameter setting, and can obtain visual testing result.The whole detection process completed by amplification instrument provided by the invention can be completed by layman, be adapted to popularize among the family that even there is inspection demand in inspection body of basic unit.
Description
Technical field
The present invention relates to scientific research, clinical examination and Food Inspection quarantine field, more particularly to genetic test field, specifically relate to
And portable constant temperature nucleic acid augmentative instrument.
Background technology
Gene amplification refers to do specific gene using archaeal dna polymerase external or invisible spectro a large amount of synthesis, it is
The chain duplication of specificity is carried out to target nucleic acids fragment using archaeal dna polymerase.Currently used technology, can be by the target of denier
DNA specifically expands more than up to a million times, so as to greatly improve the analysis to DNA molecular and detectability, which is dividing
The multi-field extensive use such as sub- biology, microbiology, medicine and science of heredity and develop rapidly.
The realization of gene amplification depends on the applicable instrument gene-amplificative instrament of detection kit and kit, base
Gene-amplification instrument provides the condition of amplified reaction and the method for result interpretation for kit, is the basic instrument for realizing gene magnification.
According to the principle of gene magnification, such as PCR, quantitative fluorescent PCR, isothermal duplication, gene-amplificative instrament is also classified into PCR instrument, constant temperature core
Sour amplification instrument etc..
Either which kind of gene-amplificative instrament, substantially all by shell, heating/cooling system, heat-conducting system, heat dissipation in its structure
A few part compositions of system, electric-control system, man-machine interface.Because its require high accuracy, high evenness temperature controlling function, be suitable for
Several genes expand the program setting ability of demand, and general gene-amplificative instrament cost is all very high, and domestic gene-amplificative instrament is all
Differed in tens of thousands of members to ten tens of thousands of yuan.In addition, because internal structure is complicated, general amplification instrument is all the reality of heavier, inconvenient movement
Room equipment is tested, can only be limited to be operated in the laboratory of specialty.
In addition, regular-PCR instrument is required for by red tape parameter setting, there is certain technology will for operating personnel
Ask.These characteristics all limit gene-amplificative instrament and popularize in the following environments：1) amateur gene magnification laboratory
Conventional environment；2) only have simple training or the operating personnel without training；3) the limited basic unit experiment focal pointe of funds；
4) regular job is for determining that target carries out the repetitive operation of identical program；5) operation of carrying instrument is needed.
The content of the invention
In order to overcome dependence and costliness of the existing gene-amplificative instrament to laboratory, operating personnel, bulky deficiency, this hair
It is bright to provide a kind of portable constant temperature nucleic acid augmentative instrument specifically for definite detection target and amplification program.The amplification instrument mainly wraps
Sample process module and amplified reaction module are included, its external form is light, can carry, it can be achieved that key start is set without other specification
Put, and visual testing result can be obtained.The whole detection process completed by the amplification instrument can be complete by layman
Into being adapted to popularize among the family that even there is inspection demand in inspection body of basic unit.
The technical solution adopted in the present invention is：
A kind of portable constant temperature nucleic acid augmentative instrument, including：
One sample process module, to carry out sample process to genetic test sample, and obtains sample treatment solution；
One amplified reaction module, to make sample treatment solution carry out amplified reaction in a reaction temperature section；
One control module, built-in control program are described to control program to control sample process module and amplified reaction
Module performs an operation for determining step.
Further, the sample process module obtains sample by being heated in a treatment temperature section to genetic test sample
Present treatment liquid.
Further, the sample process module includes：
To place the sample pool for the sample container for holding genetic test sample；
To heat the one of sample pool processing heating module；
To the cooling module to cool down to sample pool；
A processing sensing module of sample pool whether is put into judgement sample container；
To the treatment temperature sensing module of monitor sample pond temperature.
Further, the amplified reaction module includes：
To place the reaction tank of the reaction vessel for the container for holding sample treatment solution；
To heat the one of reaction tank reaction heating module；
To judge whether reaction vessel starts a reaction sensing module of amplified reaction；
To monitor the reaction temperature of reaction tank temperature sensing module.
Further, reaction tank has multiple accommodating chambers, to accommodate multiple containers for holding sample treatment solution at the same time.
Further, further include result visualization module, to by the result of amplified reaction in a manner of it can intuitively read
Present；
Indicating module, to show that operating status and prompting operating personnel perform relevant operation；
The visualization model and indicating module synchronous signal link control module.
Further, the result visualization module includes：
To irradiate a light source of the sample treatment solution for completing amplified reaction；
To observe an observation window of light source irradiation result.
Further, the indicating module includes：
One sample process indicator light, to show that sample process module heated condition and prompting add genetic test sample；
One amplified reaction indicator light, to show that amplified reaction module heated condition and prompting add sample treatment solution；
One result prompts indicator light, to prompt observation testing result.
Further, type and detection target setting of the control program according to genetic test sample.
Further, the definite step includes：
1) sample process module is preheated to heating-up temperature stabilization and is maintained at treatment temperature section, and indicating module prompting is put into base
Because detecting sample；
2) judge whether genetic test sample is put into, if so, then keeping heating-up temperature to be examined in treatment temperature section to gene
This continuous heating of test sample；
3) during heating time a to processing time, sample process module is to sample treatment solution cooling while amplified reaction module
Preheating；
4) when sample process module cools the temperature to a design temperature, sample treatment solution is taken out in indicating module prompting；
5) amplified reaction module is preheated to heating-up temperature stabilization and is maintained at reaction temperature section, and indicating module prompting is put into sample
Present treatment liquid；
6) whether judgement sample treatment fluid is put into, if so, then keeping heating-up temperature in treatment temperature section to genetic test
Sample continuous heating；
7) heating time to the amplified reaction time stops heating, and the result of amplified reaction is presented in visualization model.
Further, the treatment temperature section, reaction temperature section, processing time, the amplified reaction time is all in accordance with base
Because detecting type and the detection target setting of sample.
By taking above-mentioned technical proposal, unprofessional user can be allowed in the case of only by simple training or without training
The operation of constant temperature nucleic acid gene-amplificative instrament is completed, the operating process of whole instrument is set without complicated program parameter, it is only necessary to one
Key start can be provided with amplified reaction by internal processes in itself by instrument, its reaction result is easy to naked eyes interpretation, immediately
Obtain testing result.The equipment causes conventional environment (such as clinical department, open air, home environment, the basic unit in amateur laboratory
Hospital etc.) under carry out genetic test be possibly realized.
Brief description of the drawings
Fig. 1 is the module composition schematic diagram of portable constant temperature nucleic acid augmentative instrument in one embodiment of the invention.
Fig. 2 is the module composition signal of sample process module in portable constant temperature nucleic acid augmentative instrument in one embodiment of the invention
Figure.
Fig. 3 is the module composition signal of amplified reaction module in portable constant temperature nucleic acid augmentative instrument in one embodiment of the invention
Figure.
Fig. 4 is that the module composition of result visualization module in portable constant temperature nucleic acid augmentative instrument in one embodiment of the invention shows
It is intended to.
Fig. 5 is the module composition schematic diagram of indicating module in portable constant temperature nucleic acid augmentative instrument in one embodiment of the invention.
Embodiment
Below in conjunction with the attached drawing in the embodiment of the present invention, the technical solution in the embodiment of the present invention is carried out clear, complete
Whole description.
As shown in Figure 1, in one embodiment, there is provided a kind of portable constant temperature nucleic acid augmentative instrument, including：Sample process mould
Block, to make genetic test sample carry out sample process in a treatment temperature section, and obtains sample treatment solution；Amplified reaction mould
Block, to make sample treatment solution carry out amplified reaction in a reaction temperature section；Control module, it is built-in；One control program, it is described
Program is controlled to control sample process module and amplified reaction module to perform the operation of definite step；Result visualization module,
The result of amplified reaction to be presented in a manner of it can intuitively read；Indicating module, to show operating status and prompting behaviour
Make personnel and perform relevant operation.Visualization model and indicating module synchronous signal link control module, are held by control module control
Row relevant operation.
Wherein, program is controlled according to the type of genetic test sample and detection target setting.
Determine that step includes：
1) sample process module is preheated to heating-up temperature stabilization and is maintained at treatment temperature section, and indicating module prompting is put into base
Because detecting sample.
After being first turned on the power switch of amplification instrument, control module first implements complete machine self-test (conventional program self-test), self-test
By rear, control sample process module is begun to warm up, while allows sample process indicator light to send out a warning (in representing that heating carries out),
When target temperature is heated to, the temperature sensor meeting Real-time Feedback observed temperature set up is to control module, control module
After judging its arrival control temperature upper limit, it can stop heating, temperature can slowly decline thereafter, and when sensor feedback, its temperature reaches
When controlling lowest temperature, control module is notified that module reheats thus by dynamic control, maintains sample process module
One stationary temperature (surrounding a baseline fluctuation up and down).After arrival target temperature stablizes a bit of time (1-2min),
Control module control sample process indicator light is changed into green flicker state, notifies operator sample container can be inserted into sample
Pond.Cdna sample and the sample process agent of collection are contained in sample container.
2) judge whether genetic test sample is put into, if so, then keeping heating-up temperature to be examined in treatment temperature section to gene
This continuous heating of test sample.
After sample container is inserted into sample pool, infrared switch can be triggered, the timing of control module meeting automated execution, temperature control, hold
It is continuous that genetic test sample is heated.
3) during heating time a to processing time (such as 10 minutes), sample process module cools down at the same time sample treatment solution
Amplified reaction module preheats.
After sample process, while starting cooling module to obtaining sample treatment solution cooling, amplified reaction module is opened
Begin to preheat, ibid, warm amplified reaction indicator light takes on a red color.
4) when sample process module cools the temperature to a design temperature, sample treatment solution is taken out in indicating module prompting.
When the temperature of sample pool drops to design temperature (safe temperature), stop cooling, sample process indicator light green light dodges
Bright, prompting can take out sample container.After sample container is taken out, sample process indicator light automatic distinguishing, stops prompting.
5) amplified reaction module is preheated to heating-up temperature stabilization and is maintained at reaction temperature section, and indicating module prompting is put into sample
Present treatment liquid.
At this time, the sample treatment solution that user is manually operated in sample container is transferred to reaction vessel.Reaction vessel is generally
It is multiple.At this time, reaction tank is preheating to up to after target temperature, and observed temperature is fed back to control mould by the temperature sensor of reaction tank
Block, makes it stop heating, dynamic start-stop is so that it maintains target temperature (temperature control process of the process with sample process).In target
After temperature stabilization 1-2Min, the meeting green light flicker of amplified reaction indicator light, prompts operator to add reaction vessel.
6) whether judgement sample treatment fluid is put into, if so, then keeping heating-up temperature in treatment temperature section to genetic test
Sample continuous heating.
After reaction vessel is added, capping pond, while can trigger Hall switch immediately, starts timing, temperature control (reaction
Program), while amplified reaction indicator light is changed into red, expression is being heated.
7) heating time to the amplified reaction time (such as 40 minutes) stops heating, and amplified reaction is presented in visualization model
Result.
After the completion of response procedures, control module is automatically stopped reaction module heating, while orders unlatching light source, such as purple
The sample treatment solution of completion amplified reaction in outer light irradiation reaction vessel, while result prompting indicator light flicker, prompting can be with
Observe result.After observation, turn off the power switch.
Temperature described in the above process not refers in particular to an accurate point value or industry personnel's common cognition
With the temperature range for allowing to float and there is certain error of receiving.Above-mentioned treatment temperature section, reaction temperature section, processing
Time, type and detection goal-setting of the amplified reaction time all in accordance with genetic test sample.For example, it is directed to cervical cell DNA
Sample, it is room temperature to 100 DEG C (are, for example, 90 DEG C) that can preset treatment temperature, and predeterminable amplified reaction temperature is 65 DEG C ± 5 DEG C
In the range of a certain temperature.
Temperature and time condition is preset all in accordance with applicable sample optimum reaction condition, for special, single sample
This detection, the setting of any program parameter need not be carried out when user is operated, it is only necessary to according to the prompting of operation instruction into
Row simple operations.Operation instruction is write generally according to above-mentioned definite step, is pass on to user as teachings book.
The concrete function framework of each group into module is described as follows below：
As shown in Fig. 2, sample process module includes：Place the sample pool for the sample container for holding genetic test sample；With
To heat the one of sample pool processing heating module；To the cooling module to cool down to sample pool；Whether judgement sample container is put into
The processing sensing module of sample pool；To the treatment temperature sensing module of monitor sample pond temperature, the example according to aforementioned process
Such as it is a temperature sensor.It is, for example, infrared sensor to handle sensing module.After sample container is put into sample pool, infrared biography is triggered
Sensor, infrared sensor send trigger signal and sample are heated to control module, control module control process heating module.It is infrared
Sensor model number is for example optional：Infrared transmitting tube model：IR204-A, reception pipe PT204-6B.
Need to comparative illustration be the amplification instrument that existing laboratory uses, such as PCR instrument is most of not have gene
Sample process function, although its correlation module can realize alternating temperature, but if to carry out sample process, it is necessary to temperature tune
Required temperature is made into, it is necessary to the operation of the knowledge and specialty of specialty, and the container of general sample process and amplified reaction
The size of container also differs, and the sample cell of PCR instrument is smaller, can not be adapted to the container for placing sample process.
As shown in figure 3, amplified reaction module includes：Place the reaction of the reaction vessel for the container for holding sample treatment solution
Pond；To heat the reaction heating module of reaction tank；The reaction temperature sensing module of reaction tank temperature is monitored, is, for example, a temperature
Sensor.Judge whether reaction vessel starts the reaction sensing module of amplified reaction, reaction sensing module be, for example, trigger-type suddenly
Your sensor, identification sample process liquid container are put into reaction tank, and reaction tank closing meet reaction heating condition send it is tactile
Signal to control module, control module control heating module heating.For example optional model of Hall sensor：UHE4913G-
AE3-R substitutes model：OH34N.
Specifically, reaction tank has multiple accommodating chambers, to accommodate multiple containers for holding sample treatment solution at the same time.
As shown in figure 4, result visualization module includes：To irradiate the light source for the sample treatment solution for completing amplified reaction；
To observe the observation window of light source irradiation result.
Amplified reaction inside modules are provided with the size fit light source that standing wave is grown really, such as ultraviolet source, optical source wavelength
Fluorescent dye in corresponding amplification reaction reagent.For example, corresponding a certain fluorescent dye chooses the light source of 365nm wavelength, see
Examining the position of opening of form can be such that the position of reaction solution in reaction vessel fully exposes, and the reaction system of positive amplification exists
Aobvious yellow-green fluorescence under the excitation of 365nm ultraviolet (bottom excitation), negative reaction also have weak yellow-green fluorescence.In order to make visually
The aberration of yin and yang attribute becomes apparent from, and considers to add one layer of colour filter before form is reacted, by other jammr band mistakes outside yellow green
Filter, become apparent from yellow green.So that the identification higher of reaction result yin and yang attribute color.
As shown in figure 5, indicating module includes：Sample process indicator light, to show sample process module heated condition and
Prompting adds genetic test sample；Amplified reaction indicator light, to show that amplified reaction module heated condition and prompting add sample
Present treatment liquid；As a result indicator light is prompted, to prompt observation testing result.
One key is started shooting when the portable constant temperature nucleic acid augmentative instrument of above-described embodiment description uses, and is inserted into and is corresponded to when sample container
Processing pond when, the addition of sample container can be detected through sensor, automatically begin to operation temperature control program, sample process knot
Sample-adding is carried out by operator to operate, sample DNA liquid is added into sample process liquid container one by one, such as target detection reagent is anti-after beam
Ying Guan, when sample pool of the operator by reaction tube intercalation reaction area, amplification instrument can detect reaction tube through default sensor
Addition, automatic start gene magnification temperature control program.After amplification, the light source of reaction tank bottom automatically turns on, reaction tube
Testing result is clear and legible under light source activation.User can directly be observed by the observation window that amplification instrument relevant position opens up
Reaction result, judges yin and yang attribute.If user judges to have any problem to color, can also be by being compared with the color card enclosed
Compared with confirmation yin and yang attribute.Or can be taken pictures by mobile client, such as mobile phone to reaction result, upload pictures are to one remote
Journey server end, preset color sample or color data in remote server, pass through and carry out analysis meter to the optical data of photo
Calculate, result is judged according to preset decision content, will determine that result feeds back to mobile phone immediately, testing result is presented.Entirely
Operation requirement of the process to instrument only has the start of one key of a step.
Obviously, described embodiment is only part of the embodiment of the present invention, instead of all the embodiments.Based on this hair
Embodiment in bright, the every other implementation that those of ordinary skill in the art are obtained without making creative work
Example, belongs to the scope of protection of the invention.
Claims (10)
- A kind of 1. portable constant temperature nucleic acid augmentative instrument, it is characterised in that including：One sample process module, to carry out sample process to genetic test sample, and obtains sample treatment solution；One amplified reaction module, to make sample treatment solution carry out amplified reaction in a reaction temperature section；One control module, built-in control program are described to control program to control sample process module and amplified reaction module Perform an operation for determining step.
- 2. portable constant temperature nucleic acid augmentative instrument as claimed in claim 1, it is characterised in that the sample process module by Genetic test sample is heated and obtains sample treatment solution in one treatment temperature section.
- 3. portable constant temperature nucleic acid augmentative instrument as claimed in claim 2, it is characterised in that the sample process module includes：To place the sample pool for the sample container for holding genetic test sample；To heat the one of sample pool processing heating module；To the cooling module to cool down to sample pool；A processing sensing module of sample pool whether is put into judgement sample container；To the treatment temperature sensing module of monitor sample pond temperature.
- 4. portable constant temperature nucleic acid augmentative instrument as claimed in claim 1, it is characterised in that the amplified reaction module includes：To place the reaction tank of the reaction vessel for the container for holding sample treatment solution；To heat the one of reaction tank reaction heating module；To judge whether reaction vessel starts a reaction sensing module of amplified reaction；To monitor the reaction temperature of reaction tank temperature sensing module.
- 5. such as Claims 1-4 any one of them portable constant temperature nucleic acid augmentative instrument, it is characterised in that further include：Result visualization module, the result of amplified reaction to be presented in a manner of it can intuitively read；Indicating module, to show that operating status and prompting operating personnel perform relevant operation；The visualization model and indicating module synchronous signal link control module.
- 6. portable constant temperature nucleic acid augmentative instrument as claimed in claim 5, it is characterised in that the result visualization module bag Include：To irradiate a light source of the sample treatment solution for completing amplified reaction；To observe an observation window of light source irradiation result.
- 7. portable constant temperature nucleic acid augmentative instrument as claimed in claim 5, it is characterised in that the indicating module includes：One sample process indicator light, to show that sample process module heated condition and prompting add genetic test sample；One amplified reaction indicator light, to show that amplified reaction module heated condition and prompting add sample treatment solution；One result prompts indicator light, to prompt observation testing result.
- 8. portable constant temperature nucleic acid augmentative instrument as claimed in claim 1, it is characterised in that the control program is examined according to gene The type of test sample sheet and detection target setting.
- 9. portable constant temperature nucleic acid augmentative instrument as claimed in claim 8, it is characterised in that the definite step includes：1) sample process module is preheated to heating-up temperature stabilization and is maintained at treatment temperature section, and indicating module prompting is put into gene inspection Test sample sheet；2) judge whether genetic test sample is put into, if so, then keeping heating-up temperature in treatment temperature section to genetic test sample This continuous heating；3) during heating time a to processing time, amplified reaction module is pre- at the same time to sample treatment solution cooling for sample process module Heat；4) when sample process module cools the temperature to a design temperature, sample treatment solution is taken out in indicating module prompting；5) amplified reaction module is preheated to heating-up temperature stabilization and is maintained at reaction temperature section, and indicating module prompting is put at sample Manage liquid；6) whether judgement sample treatment fluid is put into, if so, then keeping heating-up temperature in treatment temperature section to genetic test sample Continuous heating；7) heating time to the amplified reaction time stops heating, and the result of amplified reaction is presented in visualization model.
- 10. portable constant temperature nucleic acid augmentative instrument as claimed in claim 9, it is characterised in that the treatment temperature section, reaction Temperature range, processing time, type and detection target setting of the amplified reaction time all in accordance with genetic test sample.
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| CN201711182767.0A CN107937268A (en) | 2017-11-23 | 2017-11-23 | Portable constant temperature nucleic acid augmentative instrument |
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| CN201711182767.0A CN107937268A (en) | 2017-11-23 | 2017-11-23 | Portable constant temperature nucleic acid augmentative instrument |
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Cited By (1)
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