CN105331531A - Portable virus detection device and virus detection method by polymerase chain reaction - Google Patents
Portable virus detection device and virus detection method by polymerase chain reaction Download PDFInfo
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- CN105331531A CN105331531A CN201510683374.2A CN201510683374A CN105331531A CN 105331531 A CN105331531 A CN 105331531A CN 201510683374 A CN201510683374 A CN 201510683374A CN 105331531 A CN105331531 A CN 105331531A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
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Abstract
The invention discloses a portable virus detection device by polymerase chain reaction. The portable virus detection device by polymerase chain reaction comprises a single chip microcomputer control module, a heating and cooling module and a laser detection module. According to the technical scheme, the single chip microcomputer control module controls temperatures of the heating and cooling module by controlling input and output pins of a single chip microcomputer through a PID (proportion-integration-differentiation) control algorithm, and performing data processing according to temperature data and target data transmitted by a collecting temperature sensor to control heating and cooling output power. Data obtained through the PID algorithm are outputted to pin holes in a PWM (pulse-width modulation) mode so as to control power drive of the heating and cooling module. A CPU (central processing unit) adopts 89S52 chip of an 8051 system, and a thin-film heating element and a semiconductor cooling element are adopted by a heating and cooling drive circuit. The portable virus detection device by polymerase chain reaction has the advantages that an original desktop instrument is changed into a portable pocketed instrument, and in terms of technical characteristics, the portable virus detection device can be as precise as the desktop instrument in temperature control.
Description
Technical field
The present invention relates to polymerase chain reaction, particularly relate to a kind of portable polymerase chain reaction Viral diagnosis device and detection method.
Background technology
Polymerase chain reaction (PCR, PolymeraseChainReaction) be a kind of molecular biological technology, it is used to amplify or copy section of DNA sequence to produce thousands of even up to ten thousand sections of specific dna sequences, and the process of polymerase chain reaction as shown in Figure 1.This method depends on thermal cycling, contains several sections of circulations between specified temp to provide copying of the decomposition of DNA and enzyme.Required DNA fragmentation is contained in original DNA chain.The enzyme added will be selected specific DNA fragmentation and copy.Therefore, be no matter the remains of the extinct plants and animal in fossil, historical personage, hair, skin or blood that still assailant leaves in murder case decades ago, as long as can isolate the DNA of a wee bit, just can be amplified with PCR, be compared.This is also the place of the power of " micro-evidence ".
Polymerase chain reaction be utilize DNA in vitro 95 ° of high temperature time variations Celsius can become strand, time low temperature (being often about 60 DEG C), primer is combined with the principle of strand by base pair complementarity, temperature regulating is to archaeal dna polymerase optimal reactive temperature (about 72 DEG C) again, the direction composition complementary strand of archaeal dna polymerase along phosphoric acid to five-carbon sugar (5'-3').PCR instrument based on polysaccharase manufacture is actual is exactly a temperature control device, can in denaturation temperature, and renaturation temperature, controls between elongating temperature well.Existing PCR temperature control unit mostly is table-top instrument, and not only volume is large, complex structure, and expensive, and common domestic PCR instrument is all more than ten thousand yuan.Therefore, those skilled in the art is devoted to a kind of portable polymerase chain reaction of exploitation Viral diagnosis device, and it has miniaturization, and can replace the feature of desk device.
Summary of the invention
Because the above-mentioned defect of prior art, technical problem to be solved by this invention is how by the miniaturization of existing PCR instrument device, increases portability.
For achieving the above object, the invention provides a kind of portable polymerase chain reaction Viral diagnosis device, comprise single chip control module, heating refrigerating module, laser detection module and be subject to temp controlled vessel, described single chip control module is configured to control described heating refrigerating module, and process is from the data of described laser detection module.
Further, described heating refrigerating module comprises temperature sensor, heating module, refrigerating module, the first power control module and the second power control module; Described first power control module is configured to the instruction accepting described single chip control module, controls described heating module; Described second power control module is configured to the instruction accepting described single chip control module, controls described refrigerating module.
Further, described laser detection module is configured to detect the described DNA concentration be subject in temp controlled vessel, and the data of described DNA concentration are supplied to described single chip control module.
Further, described single chip control module adopts pid algorithm to the control algolithm of described heating refrigerating module.
Further, described single chip control module adopts PWM mode to described first power control module and described second power control module.
Further, described heating module adopts film heating sheet; Described refrigerating module adopts semi-conductor cooling fin.
Further, described single chip control module adopts 8051 series monolithics.
Further, described 8051 series monolithics adopt 98S52.
Further, the P1 mouth of described 8051 series monolithics is provided with manual key, and described first power control module and described second power control module are arranged on P2 mouth, and described temperature sensor and described laser detection module installation are at P2 mouth.
Present invention also offers a kind of portable polymerase chain reaction method for detecting virus, comprising:
S1, provide as any portable polymerase chain reaction Viral diagnosis device of claim 1-9;
S2, provide target template DNA and primer;
S3, heating are subject to temp controlled vessel to 95 degree and maintain 1 minute;
S4, repeat following three step 20-35 circulations:
A) the described temp controlled vessel that is subject to of heating is to 94-98 degree, maintains 20-30 second;
B) the described temp controlled vessel that is subject to of cooling is to 50-60 degree, maintains 20-40 second;
C) the described temp controlled vessel that is subject to of heating is to 70-75 degree, maintains 15-40 second;
S5, the described temp controlled vessel that is subject to of maintenance, at 70-74 degree, maintain 5-15 minute;
24 hours are no more than by temp controlled vessel at 4-15 degree described in S6, maintenance.
Technical scheme of the present invention PCR reaction unit is implanted an integrated system, to be built into a small-sized equipment to detect specific virus.This equipment can be divided into three modules: be first single chip control module, is used for controlling heating refrigerating module and laser detection module, is also used for the data of process from laser detection module to determine the density of DNA.Next is heating refrigerating module and laser detection module.
The step required for whole PCR process of technical scheme of the present invention and the time as follows:
1. initial procedure: be heated to 95 degree and maintain 1 minute;
2.20-35 individual three step circulations:
A) sex change: be heated to 94-98 degree, maintains 20-30 second;
B) anneal: be cooled to 50-60 degree, maintain 20-40 second;
C) extend: be heated to 70-75 degree, maintain 15-40 second;
3. finally extend: maintain 70-74 degree, maintain 5-15 minute;
4. finally maintain: being maintained at 4-15, to spend the time some, but are no more than 24 hours.
The single chip control module of technical scheme of the present invention is for heating refrigerating module, control by have employed pid control algorithm to micro-chip input and output stitch, the temperature data passed back by collecting temperature sensor and target temperature carry out data processing, control to heat the output rating with cooling, thus complete temperature control.Through the data that pid algorithm draws, we output to stitch mouth to control to heat the power drive of refrigerating module by PWM mode.Technical scheme of the present invention heating cooling driving circuit adopt be film heating sheet and semi-conductor cooling fin.
The technique effect that technical scheme of the present invention can reach is that script table-top instrument is become the small portable instrument that can load pocket, but in technical characteristic, also can reach the accurate control of table-top instrument to temperature.
Be described further below with reference to the technique effect of accompanying drawing to design of the present invention, concrete structure and generation, to understand object of the present invention, characteristic sum effect fully.
Accompanying drawing explanation
Fig. 1 is the principle schematic of polymerase chain reaction;
Fig. 2 is the hardware circuit diagram of a preferred embodiment of the present invention;
Fig. 3 is the system construction drawing of a preferred embodiment of the present invention;
Fig. 4 is the pid control algorithm schematic diagram of a preferred embodiment of the present invention.
Embodiment
As shown in Figure 3, technical scheme of the present invention PCR reaction unit is implanted an integrated system, to be built into a small-sized equipment to detect specific virus.This equipment can be divided into three modules: be first single chip control module, is used for controlling heating refrigerating module and laser detection module, is also used for the data of process from laser detection module to determine the density of DNA; Next is heating refrigerating module and laser detection module.
The step required for whole PCR process of technical scheme of the present invention and the time as follows:
1. initial procedure: be heated to 95 degree and maintain 1 minute;
2.20-35 individual three step circulations:
A) sex change: be heated to 94-98 degree, maintains 20-30 second;
B) anneal: be cooled to 50-60 degree, maintain 20-40 second;
C) extend: be heated to 70-75 degree, maintain 15-40 second;
3. finally extend: maintain 70-74 degree, maintain 5-15 minute;
4. finally maintain: being maintained at 4-15, to spend the time some, but are no more than 24 hours.
So, small-sized PCR proofing unit of the present invention, if perform 20 circulations, the total time of so PCR reaction is 1+5+20 × (20+20+15)/60, is approximately 25 minutes.
The single chip control module of technical scheme of the present invention is for heating refrigerating module, control by have employed pid control algorithm to micro-chip input and output stitch, as shown in Figure 4, the temperature data passed back by collecting temperature sensor and target temperature carry out data processing, control to heat the output rating with cooling, thus complete temperature control.Through the data that pid algorithm draws, we output to stitch mouth to control to heat the power drive of refrigerating module by PWM mode.
As shown in Figure 2, the total system schematic circuit of technical scheme of the present invention, the 89S52 chip of 8051 systems that CPU adopts, wherein the P1 mouth of micro-chip is provided with manual key, heating and cooling module installation is at P2 mouth, and temperature sensor and laser detection module installation are at P2 mouth.
Technical scheme of the present invention heating cooling driving circuit adopt be film heating sheet and semi-conductor cooling fin.
The pid control algorithm of the single chip control module of technical scheme of the present invention is as follows:
/*************pid.c******************/
ucharset；
uintrout；//PIDResponse(Output)
ucharhigh_time,count＝0；
ucharnum1＝0；
ucharnum＝0；
ucharmyx＝0；
ucharsw＝1；
ucharten＝0；
/*************PID**********************************/
structPID{
uintSetPoint；//DesiredValue
uintProportion；//ProportionalConst
uintIntegral；//IntegralConst
uintDerivative；//DerivativeConst
signedintLastError；//Error[-1]
signedintPrevError；//Error[-2]
signedintSumError；//SumsofErrors
}；
structPIDspid；//PIDControlStructure
/****************pidinit*********************/
voidinit_pid()
{
high_time＝50；
spid.Proportion＝23；//SetPIDCoefficients
spid.Integral＝2；
spid.Derivative＝6；
spid.SetPoint＝set；//SetPIDSetpoint
}
/***************************PIDalgorithm**************************/
unsignedintPIDCalc(structPID*pp,unsignedintNextPoint)
{
signedintdError,Error；
Error＝pp->SetPoint-NextPoint；
pp->SumError+＝Error；
dError＝pp->LastError-pp->PrevError；
pp->PrevError＝pp->LastError；
pp->LastError＝Error；
return(pp->Proportion*Error+pp->Integral*pp->SumError+pp->Derivative*
dError)；
}
Wherein, the PWM algorithm of PID control is as follows:
/********************PIDcontrolPWM*************************/
voidduty_cycle(uintt)
{
uchars；
t＝t/100；
s＝set；
if(s>t)
{
if(s-t>0.5)
high_time＝100；
else
{
rout＝PIDCalc(&spid,t)；//PerformPIDInteration
if(high_time<＝100)
high_time＝(uchar)(rout/5)；
else
high_time＝100；
}
}
else
{
high_time＝0；
}
}
More than describe preferred embodiment of the present invention in detail.Should be appreciated that the ordinary skill of this area just design according to the present invention can make many modifications and variations without the need to creative work.Therefore, all technician in the art, all should by the determined protection domain of claims under this invention's idea on the basis of existing technology by the available technical scheme of logical analysis, reasoning, or a limited experiment.
Claims (10)
1. a portable polymerase chain reaction Viral diagnosis device, it is characterized in that, comprise single chip control module, heating refrigerating module, laser detection module and be subject to temp controlled vessel, described single chip control module is configured to control described heating refrigerating module, and process is from the data of described laser detection module.
2. portable polymerase chain reaction as claimed in claim 1 Viral diagnosis device, it is characterized in that, described heating refrigerating module comprises temperature sensor, heating module, refrigerating module, the first power control module and the second power control module; Described first power control module is configured to the instruction accepting described single chip control module, controls described heating module; Described second power control module is configured to the instruction accepting described single chip control module, controls described refrigerating module.
3. portable polymerase chain reaction as claimed in claim 1 Viral diagnosis device, it is characterized in that, described laser detection module is configured to detect the described DNA concentration be subject in temp controlled vessel, and the data of described DNA concentration are supplied to described single chip control module.
4. portable polymerase chain reaction as claimed in claim 1 Viral diagnosis device, is characterized in that, described single chip control module adopts pid algorithm to the control algolithm of described heating refrigerating module.
5. portable polymerase chain reaction as claimed in claim 2 Viral diagnosis device, is characterized in that, described single chip control module adopts PWM mode to described first power control module and described second power control module.
6. portable polymerase chain reaction as claimed in claim 2 Viral diagnosis device, is characterized in that, described heating module adopts film heating sheet; Described refrigerating module adopts semi-conductor cooling fin.
7. portable polymerase chain reaction as claimed in claim 2 Viral diagnosis device, it is characterized in that, described single chip control module adopts 8051 series monolithics.
8. portable polymerase chain reaction as claimed in claim 7 Viral diagnosis device, is characterized in that, described 8051 series monolithics adopt 98S52.
9. portable polymerase chain reaction as claimed in claim 7 Viral diagnosis device, it is characterized in that, the P1 mouth of described 8051 series monolithics is provided with manual key, described first power control module and described second power control module are arranged on P2 mouth, and described temperature sensor and described laser detection module installation are at P2 mouth.
10. a portable polymerase chain reaction method for detecting virus, is characterized in that, comprising:
S1, provide as any portable polymerase chain reaction Viral diagnosis device of claim 1-9;
S2, provide target template DNA and primer;
S3, heating are subject to temp controlled vessel to 95 degree and maintain 1 minute;
S4, repeat following three step 20-35 circulations:
A) the described temp controlled vessel that is subject to of heating is to 94-98 degree, maintains 20-30 second;
B) the described temp controlled vessel that is subject to of cooling is to 50-60 degree, maintains 20-40 second;
C) the described temp controlled vessel that is subject to of heating is to 70-75 degree, maintains 15-40 second;
S5, the described temp controlled vessel that is subject to of maintenance, at 70-74 degree, maintain 5-15 minute;
24 hours are no more than by temp controlled vessel at 4-15 degree described in S6, maintenance.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510683374.2A CN105331531A (en) | 2015-10-20 | 2015-10-20 | Portable virus detection device and virus detection method by polymerase chain reaction |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510683374.2A CN105331531A (en) | 2015-10-20 | 2015-10-20 | Portable virus detection device and virus detection method by polymerase chain reaction |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102176125A (en) * | 2011-02-22 | 2011-09-07 | 武汉馨世生物科技有限公司 | General intelligent control device for polymerase chain reaction |
| CN102559482A (en) * | 2011-12-31 | 2012-07-11 | 中山大学达安基因股份有限公司 | Gene detection device |
| CN104120077A (en) * | 2014-08-03 | 2014-10-29 | 张金木 | Real-time fluorescent DNA amplification instrument |
| CN104120078A (en) * | 2014-08-02 | 2014-10-29 | 张金木 | Real-time fluorescence PCR (Polymerase Chain Reaction) amplification instrument |
| CN104164363A (en) * | 2013-05-16 | 2014-11-26 | 北京出入境检验检疫局检验检疫技术中心 | Portable nucleic acid isothermal amplification instrument |
| CN104635801A (en) * | 2013-11-10 | 2015-05-20 | 西安扩力机电科技有限公司 | PCR (polymerase chain reaction) instrument temperature controlling system |
-
2015
- 2015-10-20 CN CN201510683374.2A patent/CN105331531A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102176125A (en) * | 2011-02-22 | 2011-09-07 | 武汉馨世生物科技有限公司 | General intelligent control device for polymerase chain reaction |
| CN102559482A (en) * | 2011-12-31 | 2012-07-11 | 中山大学达安基因股份有限公司 | Gene detection device |
| CN104164363A (en) * | 2013-05-16 | 2014-11-26 | 北京出入境检验检疫局检验检疫技术中心 | Portable nucleic acid isothermal amplification instrument |
| CN104635801A (en) * | 2013-11-10 | 2015-05-20 | 西安扩力机电科技有限公司 | PCR (polymerase chain reaction) instrument temperature controlling system |
| CN104120078A (en) * | 2014-08-02 | 2014-10-29 | 张金木 | Real-time fluorescence PCR (Polymerase Chain Reaction) amplification instrument |
| CN104120077A (en) * | 2014-08-03 | 2014-10-29 | 张金木 | Real-time fluorescent DNA amplification instrument |
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