CN104120078A - Real-time fluorescence PCR (Polymerase Chain Reaction) amplification instrument - Google Patents
Real-time fluorescence PCR (Polymerase Chain Reaction) amplification instrument Download PDFInfo
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- CN104120078A CN104120078A CN201410377161.2A CN201410377161A CN104120078A CN 104120078 A CN104120078 A CN 104120078A CN 201410377161 A CN201410377161 A CN 201410377161A CN 104120078 A CN104120078 A CN 104120078A
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- 230000003321 amplification Effects 0.000 title claims abstract description 13
- 238000003199 nucleic acid amplification method Methods 0.000 title claims abstract description 13
- 238000003752 polymerase chain reaction Methods 0.000 title abstract 4
- 238000012408 PCR amplification Methods 0.000 claims abstract description 28
- 238000001514 detection method Methods 0.000 claims abstract description 25
- 238000010438 heat treatment Methods 0.000 claims abstract description 25
- 238000005057 refrigeration Methods 0.000 claims abstract description 21
- 238000006243 chemical reaction Methods 0.000 claims abstract description 11
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims description 30
- 230000003287 optical effect Effects 0.000 claims description 22
- 229910052697 platinum Inorganic materials 0.000 claims description 15
- 230000005284 excitation Effects 0.000 claims description 11
- 238000001914 filtration Methods 0.000 claims description 11
- 239000004020 conductor Substances 0.000 claims description 6
- 230000005540 biological transmission Effects 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 4
- 238000004153 renaturation Methods 0.000 claims description 3
- 230000035945 sensitivity Effects 0.000 claims description 3
- 239000003570 air Substances 0.000 claims description 2
- 239000012080 ambient air Substances 0.000 claims description 2
- 238000004164 analytical calibration Methods 0.000 claims description 2
- 238000012937 correction Methods 0.000 claims description 2
- 230000004807 localization Effects 0.000 claims description 2
- 238000003756 stirring Methods 0.000 claims description 2
- 238000012795 verification Methods 0.000 claims description 2
- 238000000338 in vitro Methods 0.000 abstract description 3
- 238000004445 quantitative analysis Methods 0.000 abstract description 2
- 102000053602 DNA Human genes 0.000 abstract 2
- 108020004414 DNA Proteins 0.000 abstract 2
- 238000000034 method Methods 0.000 description 3
- 125000004122 cyclic group Chemical group 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000003750 conditioning effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
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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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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
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- Health & Medical Sciences (AREA)
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- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
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- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biomedical Technology (AREA)
- Engineering & Computer Science (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention relates to a real-time fluorescence PCR (Polymerase Chain Reaction) amplification instrument which consists of a heating/refrigeration module, a sample tube carrying disc component, a detection module, a sensor and a controller. The amplification instrument is used for amplifying specific DNA (Deoxyribose Nucleic Acid) segments in vitro and used for detecting the change of each circulation amplification product in PCR amplification reaction in real time according to change of fluorescence signals, and thus quantitative analysis can be performed.
Description
Technical field:
The present invention relates to a kind of real-time fluorescence PCR amplification instrument, it forms by heating refrigeration module, sample hose load plate assembly, detection module, sensor and controller.For the specific DNA fragmentation that increases in vitro, utilize the variation of fluorescent signal simultaneously, detect in real time the variation of the amount of each cyclic amplification product in pcr amplification reaction, in order to carry out quantitative analysis.
Prior art:
The semiconservative replication of DNA is organic evolution and the important channel of going down to posterity.People find in testing in vitro, and DNA sex change can occur when high temperature and unwinds, and after temperature reduces, can renaturation become two strands again.Therefore, coordinate biological means by temperature variation, to control the denature and renature of DNA, and the circulation that repeatedly repeats " sex change unwind-anneal-synthesize extension " just can be with the geometricprogression specific genes that increase in a large number.In the evolution of round pcr, pcr amplification program has been simplified in the discovery of heat-stable DNA polymerase and the automatization of temperature cycle greatly.Fluorescent quantitative PCR technology is to add fluorophor in PCR reaction system, and the accumulation along with its fluorescence signal intensity of recycle of PCR reaction, can realize the whole PCR process of Real-Time Monitoring.
At present, pcr amplification instrument generally has Four types, respectively regular-PCR amplification instrument, fluorescent PCR amplification instrument, grads PCR amplification instrument and situ PCR instrument, their principles are similar, but due to accurate temperature, controlling is the key that determines PCR reaction success, therefore in different application of having nothing in common with each other in aspect such as apparatus structure, heating refrigeration mode and fluoroscopic examination modes.The application of pcr amplification instrument has: the molecular diagnosis of infectivity, heredopathia and malignant tumour and research, paternity test, clone gene etc.
Summary of the invention:
After most pcr amplification instrument amplifications finish, do not detect, it is to be completed by miscellaneous equipment that the terminal product of its amplified reaction detects.Fluorescent PCR amplification instrument comprises real-time fluorescence PCR amplification instrument and common fluorescent PCR amplification instrument, and the latter is detected by this amplification instrument after amplification finishes.Real-time fluorescence PCR amplification instrument is to detect at amplification procedure, and it need solve detection mode and detection speed problem.Fluorescent PCR amplification instrument is to adopt very low temperature CCD imaging at present, this system is once to multiple spot imaging, but detection sensitivity low accuracy is poor, the employing optical system also having is scanning samples detection one by one, but its scanning system complexity involves great expense and detection speed is slow, therefore the accurate amplification detection in real time for most of a small amount of samples is very uneconomic.The present invention introduces a kind of real-time fluorescence PCR amplification instrument, and it forms by heating refrigeration module, sample hose load plate assembly, detection module, platinum resistance thermometer sensor, and controller.It is characterized in that heating refrigeration module and be comprised of Electrothermal ring, refrigerator, scatterer and electric fan, have a heating refrigerator compartment between Electrothermal ring and refrigerator, Electrothermal ring and refrigerator power variable, for to its heated with ambient air refrigeration.Refrigerator is equipped with conductor refrigeration sheet, each semi-conductor refrigerating sheet can be connected into series-parallel system to be applicable to its power and service voltage demand, the hot side of conductor refrigeration sheet contacts with scatterer for outwards heat radiation, huyashi-chuuka (cold chinese-style noodles) contacts with heat-conducting plate, and realizes change cooling power by changing conductor refrigeration sheet supply current.The interstitial hole of the hot and cold air of Electrothermal ring and refrigerator by heating refrigerator compartment, Electrothermal ring stirs evenly under backward and is pushed to the sample in sample hose load plate through electric fan, and this structure has been isolated the inhomogeneous hot gas flow making progress makes sample temperature more even.Sample hose load plate assembly is comprised of sample hose load plate, stepper-motor and transmission shaft.The outer circle of sample hose load plate is the hole for placing sample hose along circumference equal distribution, in the outer circled in the hole of sample hose, bore a pilot hole for laser-correlation locating device is installed, No. 1 hole with identification sample hose, when No. 1 hole of sample hose enters measuring position, the optical signal of laser diode transmitting is just received by light Receiver by pilot hole, thereby starting detects, and drive sample hose load plate to rotate by stepper-motor, sequentially the sample in each sample hose of detection and localization.Adopt its positioning precision of laser-correlation locating device high, light Receiver in the laser diode of laser-correlation locating device in launching device and location receiving trap forms, laser diode rated output is got and is not more than 0.4mW, and adopting the laser diode pulse power that dutycycle is little is therefore foolproof.Transmission shaft two ends are fixed with axle and the sample hose load plate of stepper-motor respectively, for transmitting stepper-motor corner.In the outside of the sample hose near sample hose load plate, be uniformly distributed the sensor of 3 platinum resistance thermometer sensor,s, the compensating resistance of platinum resistance thermometer sensor, adopts accurate wire-wound resistor, 3 to 4 accurate wire-wound resistors are housed simultaneously in controller, its resistance value sets near temperature spot temperature according to DNA sex change, renaturation and three links of extension and looks into platinum resistance thermometer sensor, calibration table and obtain, due to the different therefore accurate wire-wound resistors how of its temperature of reaction of different design of primers, the temperature value record that accurate wire-wound resistor is corresponding is kept in the non-volatile memory of controller, for instrument calibration temperature.Described correction is to carry out error check once after each start, and it realizes verification by the accurate wire-wound resistor access of single multichannel analog electronic switch timesharing gating input signal amplifying circuit II.Pcr amplification instrument is 3 platinum resistance thermometer sensor, measured temperatures in working order, should be within the scope of temperature error between the sample hose allowing, timing gets each platinum resistance thermometer sensor, thermometric error by timing, and the shown temperature of accurate wire-wound resistor with described in be kept in non-volatile memory should displays temperature poor, by controller, platinum resistance thermometer sensor, measured temperature is compensated.Detection module is comprised of excitation light source and detection receiving trap.Excitation light source is by LED photodiode, reflection cup, optical filtering I and excite isolating cylinder to form, monochromatic LED photodiode price is low, less energy consumption, life-span are long, but need different LED photodiodes to join again optical filtering, could realize better different excitation wavelengths, adopt reflection cup to make light more concentrated.Detecting receiving trap forms by receiving isolating cylinder, optical filtering II and photomultiplier, excitation light source and detection receiving trap are placed in outside heating chamber, to reduce heat source fluctuations to transmitting and receiving the impact of sensitivity, and all the isolating cylinder by light transmits and receives optical signal to sample in heating chamber, isolating cylinder and optical filtering are all for getting rid of the impact of surround lighting and useless optical signal, in order to improve light accuracy of detection.Controller is comprised of single multichannel analog electronic switch, input signal amplifying circuit I, input signal amplifying circuit II, input modulate circuit I, input modulate circuit II, input multiselect one switch, A/D change-over circuit, micro-chip, RS232 circuit, heating power supply, stepper motor driving circuit and refrigerator power source circuit.Wherein in input signal amplifying circuit I, comprise by the precision operational-amplifiers such as high-precision LT1014 or OPA121 it and input modulate circuit I and forms dedicated optical signal amplification circuit, the temperature millivolt signal amplifying circuit that input signal amplifying circuit II is comprised of OP07 and input modulate circuit II and form.RS232 circuit is for communicating by letter with upper computer, and it,, by sending Computer Analysis research after fluorescent signal collection and processing, draws the real-time results of quantification.
Under the controller control therein of described a kind of real-time fluorescence PCR amplification instrument, the temperature setting according to PCR reaction link is first with high-power intensification or cooling, after reaching the temperature of setting, again with low power heating or refrigeration, temperature controlled return difference is remained within the scope of the temperature control precision of pcr amplification permission, and make pcr amplification instrument or be cooled to every step to react required precise temp by each sample hose heating, the variation of recycling fluorescent signal detects the variation of the amount of each cyclic amplification product in pcr amplification reaction in real time, but its detection speed must guarantee that each circulation has detected all samples on sample hose load plate.Described heating refrigeration module is installed in the lid of pcr amplification instrument, opens this lid and can load and unload sample hose.
Accompanying drawing explanation:
Fig. 1 is a kind of structural representation of real-time fluorescence PCR amplification instrument.
Fig. 2 is a kind of formation block diagram of real-time fluorescence PCR amplification instrument.
Embodiment:
A kind of structural representation of real-time fluorescence PCR amplification instrument as shown in Figure 1.The heating refrigeration module that it is comprised of Electrothermal ring 3, refrigerator 2, scatterer 1 and electric fan 4, the sample hose load plate assembly being formed by sample hose load plate 14, stepper-motor 11 and transmission shaft 13, the detection module being formed by excitation light source 8 and detection receiving trap 10, and formed by platinum resistance thermometer sensor, 16, controller 12, shell 18 and lid 19.Electrothermal ring 3 adopts electrothermal tube heating, and refrigerator 2 adopts conductor refrigeration sheet.Wherein the outer circle of sample hose load plate 14 is used for placing sample hose 6 along 24 holes of circumference equal distribution, and heat lid 5 is installed on sample hose 6.In the outer circled in the hole of sample hose 6, bore a pilot hole for laser-correlation locating device is installed, this laser-correlation locating device consists of the laser diode of launching device 17 and the light Receiver in the receiving trap 15 of location, to identify No. 1 hole of sample hose load plate 14.By LED photodiode, reflection cup, optical filtering I with excite isolating cylinder 7 to form excitation light source 8, detecting receiving trap 10 forms by receiving isolating cylinder 9, optical filtering II and photomultiplier, the reflection cup that reflection cup can adopt LED LED torch to use, excitation light source 8 utilizes the wall that excites isolating cylinder and reception isolating cylinder to pass reaction chamber to transmit and receive optical signal to sample hose 6 with detection receiving trap 10, and it and optical filtering are all for getting rid of the impact of surround lighting and useless optical signal.In controller 12 by single multichannel analog electronic switch, and the temperature millivolt signal amplifying circuit being formed by input signal amplifying circuit II, input conditioning II, and nurse one's health the light input signal amplifying circuit that I forms by input signal amplifying circuit I, input.They send A/D change-over circuit, micro-chip to process after input multiselect one switching gate.The temperature millivolt signal amplifying circuit that wherein special-purpose light input signal amplifying circuit is formed and is comprised of OP07 by high precision LT1014 amplifying circuit.RS232 circuit, heating power supply, refrigeration power supply and stepper-motor 11 driving circuits are also set in controller 12.Wherein RS232 circuit is for communicating by letter with upper computer.
Claims (2)
1. a real-time fluorescence PCR amplification instrument, it is by heating refrigeration module, sample hose load plate assembly, detection module, platinum resistance thermometer sensor, and controller form, it is characterized in that heating refrigeration module by Electrothermal ring, refrigerator, scatterer and electric fan form, between Electrothermal ring and refrigerator, there is a heating refrigerator compartment, Electrothermal ring and refrigerator power variable, be used for its heated with ambient air refrigeration, refrigerator is equipped with conductor refrigeration sheet, change conductor refrigeration sheet supply current and realize change cooling power, the hot and cold air of Electrothermal ring and refrigerator is by heating refrigerator compartment, the interstitial hole of Electrothermal ring stirs evenly under backward and is pushed to the sample in sample hose load plate through electric fan, this structure has been isolated the inhomogeneous hot gas flow making progress makes sample temperature more even, sample hose load plate assembly is by sample hose load plate, stepper-motor and transmission shaft form, the hole of sample hose load plate equal distribution for placing sample hose, and laser-correlation locating device is installed, No. 1 hole with identification sample hose, when No. 1 hole of sample hose enters measuring position, the optical signal of laser diode transmitting is just received by light Receiver by pilot hole, thereby starting detects, and drive sample hose load plate to rotate by stepper-motor, the sample in each sample hose of detection and localization sequentially, adopt its positioning precision of laser-correlation locating device high, light Receiver in the laser diode of laser-correlation locating device in launching device and location receiving trap forms, laser diode rated output is got and is not more than 0.4mW, adopt the little laser diode pulse power of dutycycle, transmission shaft two ends are fixed with axle and the sample hose load plate of stepper-motor respectively, be used for transmitting stepper-motor corner, in the outside of the sample hose near sample hose load plate, be uniformly distributed the sensor of 3 platinum resistance thermometer sensor,s, the compensating resistance of platinum resistance thermometer sensor, adopts accurate wire-wound resistor, 3 to 4 accurate wire-wound resistors are housed simultaneously in controller, its resistance value is according to DNA sex change, near the temperature spot that renaturation and three links of extending set temperature is looked into the acquisition of platinum resistance thermometer sensor, calibration table, the temperature value record that accurate wire-wound resistor is corresponding is kept in the non-volatile memory of controller, for instrument calibration temperature, described correction is to carry out error check once after each start, it realizes verification by the accurate wire-wound resistor access of single multichannel analog electronic switch timesharing gating input signal amplifying circuit II, timing gets each platinum resistance thermometer sensor, thermometric error by timing, and the shown temperature of accurate wire-wound resistor with described in be kept in non-volatile memory should displays temperature poor, by controller, platinum resistance thermometer sensor, measured temperature is compensated, detection module is comprised of excitation light source and detection receiving trap, excitation light source is by LED photodiode, reflection cup, optical filtering I and excite isolating cylinder to form, need different LED photodiodes to join again optical filtering and could realize better different excitation wavelengths, adopt reflection cup to make light more concentrated, detect receiving trap by receiving isolating cylinder, optical filtering II and photomultiplier form, excitation light source and detection receiving trap are placed in outside heating chamber, to reduce heat source fluctuations to transmitting and receiving the impact of sensitivity, and all the isolating cylinder by light transmits and receives optical signal to sample in heating chamber, isolating cylinder and optical filtering are all for getting rid of the impact of surround lighting and useless optical signal, in order to improve light accuracy of detection, it is under controller is controlled that temperature is controlled, the temperature setting according to PCR reaction link is first with high-power intensification or cooling, after reaching the temperature of setting, again with low power heating or refrigeration, temperature controlled return difference is remained within the scope of the temperature control precision of pcr amplification permission.
2. a kind of real-time fluorescence PCR amplification instrument according to claim 1, characterized by further comprising:
It is by heating refrigeration module, sample hose load plate assembly, detection module, platinum resistance thermometer sensor, and controller form, its middle controller is by single multichannel analog electronic switch, input signal amplifying circuit I, input signal amplifying circuit II, input modulate circuit I, input modulate circuit II, input multiselect one switch, A/D change-over circuit, micro-chip, RS232 circuit, heating power supply, stepper motor driving circuit and refrigerator power source circuit form, wherein in input signal amplifying circuit I, comprise by the precision operational-amplifiers such as high-precision LT1014 or OPA121 it and input modulate circuit I and form dedicated optical signal amplification circuit, the temperature millivolt signal amplifying circuit that input signal amplifying circuit II is comprised of OP07 and input modulate circuit II thereof form.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105331531A (en) * | 2015-10-20 | 2016-02-17 | 上海交通大学 | Portable virus detection device and virus detection method by polymerase chain reaction |
| CN106047685A (en) * | 2016-05-18 | 2016-10-26 | 崔景香 | Fluorescent quantitative PCR (polymerase chain reaction) instrument |
| CN106929388A (en) * | 2015-12-31 | 2017-07-07 | 苏州百源基因技术有限公司 | A kind of real-time fluorescence quantitative PCR instrument |
| CN107058090A (en) * | 2017-04-27 | 2017-08-18 | 滨江华康（北京）生物科技有限公司 | A kind of real-time fluorescence quantitative PCR gene magnification detector |
| CN107164564A (en) * | 2017-07-05 | 2017-09-15 | 新疆畜牧科学院兽医研究所（新疆畜牧科学院动物临床医学研究中心） | One kind is used to detect canine parainfluenza virus kit |
| CN108139440A (en) * | 2015-03-30 | 2018-06-08 | 精工爱普生株式会社 | Electronic component handling apparatus and electronic component inspection device |
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| EP1541237A2 (en) * | 2003-12-10 | 2005-06-15 | Samsung Electronics Co., Ltd. | Polymerase chain reaction (pcr) module and multiple pcr system using the same |
| CN202881280U (en) * | 2012-11-15 | 2013-04-17 | 王波 | Real-time fluorescent quantitative PCR (polymerase chain reaction) instrument |
| KR101302353B1 (en) * | 2012-02-29 | 2013-09-06 | 케이맥(주) | Automatic analysis device of molecular diagnosis for performing both real-time pcr detecting quantitatively gene and dna microarray for gene analysis |
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2014
- 2014-08-02 CN CN201410377161.2A patent/CN104120078B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1541237A2 (en) * | 2003-12-10 | 2005-06-15 | Samsung Electronics Co., Ltd. | Polymerase chain reaction (pcr) module and multiple pcr system using the same |
| KR101302353B1 (en) * | 2012-02-29 | 2013-09-06 | 케이맥(주) | Automatic analysis device of molecular diagnosis for performing both real-time pcr detecting quantitatively gene and dna microarray for gene analysis |
| CN202881280U (en) * | 2012-11-15 | 2013-04-17 | 王波 | Real-time fluorescent quantitative PCR (polymerase chain reaction) instrument |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108139440A (en) * | 2015-03-30 | 2018-06-08 | 精工爱普生株式会社 | Electronic component handling apparatus and electronic component inspection device |
| CN105331531A (en) * | 2015-10-20 | 2016-02-17 | 上海交通大学 | Portable virus detection device and virus detection method by polymerase chain reaction |
| CN106929388A (en) * | 2015-12-31 | 2017-07-07 | 苏州百源基因技术有限公司 | A kind of real-time fluorescence quantitative PCR instrument |
| CN106047685A (en) * | 2016-05-18 | 2016-10-26 | 崔景香 | Fluorescent quantitative PCR (polymerase chain reaction) instrument |
| CN106047685B (en) * | 2016-05-18 | 2018-06-01 | 崔景香 | A kind of fluorescence quantitative PCR instrument |
| CN107058090A (en) * | 2017-04-27 | 2017-08-18 | 滨江华康（北京）生物科技有限公司 | A kind of real-time fluorescence quantitative PCR gene magnification detector |
| CN107164564A (en) * | 2017-07-05 | 2017-09-15 | 新疆畜牧科学院兽医研究所（新疆畜牧科学院动物临床医学研究中心） | One kind is used to detect canine parainfluenza virus kit |
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| CN104120078B (en) | 2015-12-02 |
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Address after: 230000 A6, road, Fuyang Road, Luyang Industrial Zone, Hefei, Anhui Patentee after: Anhui Anlong Gene Technology Co Ltd Address before: 230000 A6, road, Fuyang Road, Luyang Industrial Zone, Hefei, Anhui Patentee before: Anhui Anlong gene limited medical examination |