CN209193976U - A kind of full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions - Google Patents

A kind of full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions Download PDF

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CN209193976U
CN209193976U CN201821502049.7U CN201821502049U CN209193976U CN 209193976 U CN209193976 U CN 209193976U CN 201821502049 U CN201821502049 U CN 201821502049U CN 209193976 U CN209193976 U CN 209193976U
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sample
nucleic acid
track
reagent
detection
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刘子杰
段勇
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First Affiliated Hospital of Kunming Medical University
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First Affiliated Hospital of Kunming Medical University
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Abstract

The utility model discloses a kind of full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions, is related to technical field of medical examination.The full-automatic nucleic acids instrument, including nucleic acid-extracting apparatus, detection device and controller, nucleic acid-extracting apparatus are connected with detection device and are equipped with sealing cover, and detection device side installation controller simultaneously connects to power supply.The full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions of the utility model, can realize the continuous detection of sample of nucleic acid, the influence of not examined batch, and detection process whole-course automation can reduce technical threshold, be conducive to this technology and promote to base.

Description

A kind of full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions
Technical field
The utility model relates to technical field of medical examination, are specifically related to a kind of based on the full-automatic of isothermal amplification reactions Nucleic acid amplification assays instrument.
Background technique
Nucleic acid (Ribonucleic RNA, DNA DNA) is the basic unit for representing life entity hereditary feature, nucleic acid Detection is to carry out advanced biological detection on a molecular scale, compared to traditional morphologic detection, cytology detection, immunology inspection The remarkable advantages such as survey etc. more sensitivity, specificity are widely used in hepatitis, AIDS in the detection of clinical position amplifying nucleic acid in recent years Etc. serious infectious diseases;The emerging infectious diseases such as SARS, bird flu, hand-foot-and-mouth disease;The gene diagnosis and polygenes phase of genetic disease The chronic disease detection and diagnosis of pass.Detection of nucleic acids includes qualitative PCR (Polymerase Chain Reaction, polymerase chain Formula reaction), real-time fluorescence quantitative PCR (Real time PCR, QPCR), molecule hybridization, the skills such as genetic chip and gene sequencing Art is, it can be achieved that gene qualitative and quantitative analysis.The detections such as molecule hybridization, genetic chip and gene sequencing, and the premise of these technologies It is the amplification for realizing target gene, only just can be carried out corresponding detection after the amplification of realization target gene.
In addition, detection of nucleic acids need to have efficient and high-purity method for extracting nucleic acid.Existing nucleic acid extraction is substantially adopted With paramagnetic particle method, improvement is carried out with after surface modification with surface of the nanotechnology to superparamagnetic nano particle, is prepared into super suitable Magnetic silicon oxide nanometer magnetic bead.This magnetic bead can specifically be identified with nucleic acid molecules on micro interface and efficiently be combined.Benefit With the superparamagnetism of silica nanosphere, in the work of Chaotropic salt (guanidine hydrochloride, guanidinium isothiocyanate etc.) and externally-applied magnetic field Under, can from blood, animal tissue, food, pathogenic microorganism equal samples DNA and RNA separate, can be applicable to clinic Medical diagnosis on disease, transfusion safety, Forensic Identification, nucleic acid samples build library, environmental microorganism detection, food safety detection, molecule life The multiple fields such as object research.
The traditional PCR technique that three temperature constantly recycle is depended in clinical and research work at present, this technology is only Can carry out in batches, be difficult to add other detection samples when amplified reaction operation, at the same the method for nucleic acid extraction also because The influence of round pcr is all into batch and extracts, therefore art methods have following defects that
1. traditional PCR method based on three temperature changes when detection can not continuous sample introduction, to detection sample Batch is restricted, can not achieve the continuous detection of automation, does not meet the demand of the detection of clinical samples.
2. traditional PCR method is higher to temperature control requirement, and need to realize the accurate elevating control of temperature, therefore to wanting Equipment is asked to have degree of precision, therefore equipment is relatively expensive, is unfavorable for Technique Popularizing.
3. temperature change has an impact to the performance of enzyme in normal PCR detection method, with the extension of reaction time the work of enzyme Property constantly decline, influence reaction sensibility and specificity.
4. carrying out the biological sample of detection of nucleic acids, there are diversity, at the same because testing goal it is different required for target not yet Together, some reaction templates need DNA and some then need RNA, and different samples and the method for extracting DNA or RNA are all different, And the method that nucleic acid extraction product currently on the market is all made of batch extracting, i.e., multiple same type samples are extracted every time, or only DNA or RNA can be extracted respectively, therefore restricted to the speed of entire detection reaction.
5. it is more that traditional PCR technique is related to link, inspection result quality just can guarantee after needing professional trained, because This technical threshold is higher, is unfavorable for the popularization of technology.
Utility model content
In view of the above-mentioned problems existing in the prior art, the utility model provide it is a kind of based on isothermal amplification reactions it is complete from Dynamic nucleic acid amplification assays instrument can realize the continuous detection of sample of nucleic acid, and the influence of not examined batch, detection process whole process is certainly Dynamicization can reduce technical threshold, be conducive to this technology and promote to base.
To realize above-mentioned technical purpose and the technique effect, the utility model is achieved through the following technical solutions:
A kind of full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions, including nucleic acid-extracting apparatus, detection device And controller, nucleic acid-extracting apparatus are connected with detection device and are respectively equipped with gas-tight silo, detection device and extraction element respectively with Controller simultaneously connects to power supply;
The nucleic acid-extracting apparatus includes that sample sample introduction track, reagent track, nucleic acid extraction unit and sample go out sample track, The injection port of the sample sample introduction track connects specimen holder propeller, and rear side installs sample blending device, and connects reagent track, Reagent track two sides separately design the reagent suction needle and sample suction needle controlled by mechanical arm, and are correspondingly arranged extraction reagent Storehouse, TIP storehouses and waste holder, reagent track end connect each nucleic acid extraction unit by branching agent track, and the nucleic acid mentions Taking is reagent track among unit, and track two sides are nucleic acid extraction position, the magnetic of the upper layer installation specification arrangement of nucleic acid extraction unit Stick is simultaneously controlled by bar magnet moving arm, and nucleic acid extraction unit rear end connection sample goes out sample track, and sample track end is after extracting out Mark product storage area;
The detection device includes reaction tray and ccd image sensor (CCD) device, in the reaction tray Equipped with multiple reaction cups, and design sample outlet and detection reagent sample introduction rail, sample exhaust port and detection reagent sample introduction track Mechanical gripper is designed with reaction tray connecting pin, sample exhaust port is connected sample waste storage storehouse, pacified on detection reagent sample introduction track Sample bottle capper is filled, liquid injection port and temperature detection mouth are designed in reaction tray, CCD device is used for fluorescence detection.
Further, the nucleic acid-extracting apparatus upper end connects TIP loading channels and reagent storage storehouse, the nucleic acid mention Take unit and detection unit that ultraviolet lamp is installed.
Further, TIP storehouses in the detection device, agent bin and the sample suction needle controlled by mechanical arm and Propeller;
Further, exhaust equipment is equipped on rear side of nucleic acid-extracting apparatus and nucleic acid detection apparatus;
The agent bin is refrigeration warehouse.
A kind of full-automatic nucleic acid amplification assays instrument control system based on isothermal amplification reactions, including controller, the control Device processed includes detection unit, control unit, execution unit and display unit, and detection unit includes the bar code reading of sample introduction track Device, reaction tray temperature measuring probe, control unit and display unit are the main control chip and display for being mounted on controller, are executed Unit includes the control mechanical arm of reagent needle and specimen needle, sample sample introduction track, reagent track, sample goes out sample track, bar magnet moves Swing arm and the handgrip of detection device, mechanical arm;Barcode reader detection signal feeds back to control unit, and control unit driving executes Sample is sent to nucleic acid extraction unit by the sample sample introduction track of element, and drive control mechanical arm, reagent track and bar magnet move Swing arm carries out nucleic acid extraction, and driving sample goes out sample track and send mark product to storage area after the completion of extracting, and drives detection device later Handgrip, mechanical arm detected.
Controller control specimen needle, reagent needle and bar magnet do not conflict, that is, can also be other extractions in nucleic acid extraction Position reagent adding and sample again.
A method of the nucleic acid extraction and detection of the full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions, including Following steps:
Specimen holder to be checked send to injection port to wait and be loaded through sample sample introduction track, and through barcode reader identification sample letter Breath, the sample if necessary to mixing are then loaded after rotation mixes in mark product evenly mixing device again, go out sample through sample after sample-adding completion Sample storage area after track is sent to detection;
It extracts reagent and pushes to reagent track from agent bin, and add other additional now with reagent;
Sample needle is drawn sample and is added in extraction reagent;
Reagent track will extract reagent and be pushed to sample extraction position and fixation;
The corresponding bar magnet for extracting position completes nucleic acid extraction work in corresponding position, and reagent is pushed to detection after having extracted Storehouse, and close the door of extraction element;
It extracts reagent and pushes to detection device sample-adding position, detection reagent storehouse will test reagent and push to reagent track, sample Needle draw extract nucleic acid be added detection reagent in, extract reagent by track push to inspection after sample storage warehouse;
Reagent transparent plastic after sample-adding is covered and is sent into detection device reaction tray and detects;Reaction tray be it is centrifugal, Reaction system can be centrifuged and be mixed, can constantly resetted during the reaction, to carry out fluorescence detection;
It keeps temperature constant in detection process, and fluorescence detection is being carried out by CCD within a certain period of time;
Sample exhaust port is pushed out reaction tray after inspection after having detected.
The utility model has the beneficial effects that
1. the detection of nucleic acid quantification, qualitative, Genotyping and gene pleiomorphism can be carried out.
2. the continuous detection of sample of nucleic acid can be realized, the influence of not examined batch;
3. independent extract sample, it can be achieved that a variety of different type samples extract simultaneously, DNA or RNA can also be extracted simultaneously;
4. integrated nucleic acid extraction and detection of nucleic acids reduce the requirement of lab space in one;
5. pollution of nucleic acid risk can utmostly be reduced by being equipped with ultraviolet lamp and exhaust equipment;
Detection process whole-course automation can reduce technical threshold, be conducive to this technology and promote to base.
Certainly, any product for implementing the utility model does not necessarily require achieving all the advantages described above at the same time.
Detailed description of the invention
In order to illustrate more clearly of the technical solution of the utility model embodiment, make required for being described below to embodiment Attached drawing is briefly described, it should be apparent that, the drawings in the following description are merely some embodiments of the present invention, For those of ordinary skill in the art, without creative efforts, it can also be obtained according to these attached drawings Other attached drawings.
Fig. 1 is the structural schematic diagram described in the utility model embodiment for nucleic acid extraction and the integration apparatus of detection;
Fig. 2 is the structural schematic diagram one of nucleic acid-extracting apparatus described in the utility model embodiment;
Fig. 3 is the structural schematic diagram two of nucleic acid-extracting apparatus described in the utility model embodiment;
Fig. 4 is the floor map of nucleic acid extraction unit described in the utility model embodiment;
Fig. 5 is the positive structure schematic of nucleic acid-extracting apparatus described in the utility model embodiment;
Fig. 6 is the structural schematic diagram on nucleic acid extraction unit upper layer described in the utility model embodiment;
Fig. 7 is the structural schematic diagram of detection device described in the utility model embodiment;
The floor map of detection device described in the utility model embodiment of the position Fig. 8;
The structural schematic diagram of reaction tray described in the utility model embodiment of the position Fig. 9;
In attached drawing, parts list represented by the reference numerals are as follows:
1- nucleic acid-extracting apparatus, 2- detection device, 3- controller, 4- injection port, 5- sample introduction track, 6- sample blending dress It sets, 7- extraction agent bin, 8-TIP storehouses, 9- nucleic acid extraction unit, 10- bar magnet moving arm, 11- goes out sample track, and 12- mark product are deposited Put area, 13- barcode reader, 14- sample needle, 15- extracts reagent suction needle, 16- reagent track, 17- branching agent track, 18- waste holder, 19- bar magnet, 20- ultraviolet lamp, 21- extraction position, 22-TIP loading channels, 23- reagent storage storehouse, 24- are discarded TIP box, 25- reaction tray, 26- sample exhaust port, 27- detection detection reagent sample introduction rail, 28- propeller, 29- sample storage warehouse, 30- bottle capper, 31- reaction cup, 32- liquid injection port, 33- temperature detection mouth, 34- exhaust equipment, 35- reaction reagent storehouse, 36-CCD device.
Specific embodiment
The following will be combined with the drawings in the embodiments of the present invention, carries out the technical scheme in the embodiment of the utility model Clearly and completely describe, it is clear that the described embodiments are only a part of the embodiments of the utility model, rather than whole Embodiment.Based on the embodiments of the present invention, those of ordinary skill in the art are without creative efforts All other embodiment obtained, fall within the protection scope of the utility model.
Embodiment 1
As shown in figures 1-8
A kind of full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions, including nucleic acid-extracting apparatus 1, detection device 2 and controller 3, nucleic acid-extracting apparatus 1 is connected with detection device 2 and is equipped with gas-tight silo, and controller 3 is installed in 2 side of detection device And it connects to power supply;
The nucleic acid-extracting apparatus 1 includes sample introduction track 5, reagent rail 16, nucleic acid extraction unit 9 and sample track out 11, the injection port of the sample introduction track 5 connects specimen holder propeller, and rear side installs sample blending device 6, and connects reagent track Road 16,16 two sides of reagent rail separately design the reagent needle controlled by mechanical arm and specimen needle 14, and are correspondingly arranged extraction The storehouse agent bin 7, TIP 8 and waste holder, 16 end of reagent rail connect nucleic acid extraction unit by branching agent rail 17 9, it is reagent rail 16 among the nucleic acid extraction unit 9, two sides are to extract position 21, the upper layer installation of nucleic acid extraction unit 9 The bar magnet 19 of specification arrangement is simultaneously controlled by bar magnet moving arm, and 9 rear end of nucleic acid extraction unit connects out sample track 11, out sample track 11 ends are the sample holding region 12 after extracting;
The detection device 2 includes the CCD device 36 of reaction tray 25 and ccd image sensor, described anti- It should coil and multiple reaction cups 31 are housed on 25, and design sample outlet 26 and detection reagent sample introduction rail 27,26 He of sample exhaust port Handgrip is designed in detection reagent sample introduction rail 27 and 25 connecting pin of reaction tray, and sample exhaust port 26 connects sample waste storage storehouse, detection Sample bottle capper 30 is installed on reagent sample introduction track 27, designs liquid injection port 32 and temperature detection mouth 33, CCD in reaction tray 25 Device 36 is used for fluorescence detection.
The bottom of the nucleic acid-extracting apparatus 1 is discarded TIP box 24, and upper end connects TIP channels 22 and reagent storage storehouse 23, ultraviolet lamp 20 is installed on 9 position of nucleic acid extraction unit and nucleic acid detection apparatus right side.
It TIP storehouses 8 in the detection device 2, reaction reagent storehouse 35 and the specimen needle 14 controlled by mechanical arm and pushes away Into device;
35, reaction reagent storehouse refrigeration warehouse.
A kind of full-automatic nucleic acid amplification assays instrument control system based on isothermal amplification reactions, including controller, the control Device processed includes detection unit, control unit, execution unit and display unit, and detection unit includes that the bar code of sample sample introduction track is read Device, reaction tray temperature measuring probe are read, control unit and display unit are the main control chip (AT89C51 of installation on the controller Singlechip chip) and display, execution unit include the control mechanical arm of reagent needle and specimen needle, sample introduction track, reagent track, Sample goes out sample track, the handgrip of bar magnet moving arm and detection device, mechanical arm;It is single that barcode reader detection signal feeds back to control Mark product are sent to nucleic acid extraction unit by the sample sample introduction track of member, control unit driving executive component, and drive control is mechanical Arm, reagent track and bar magnet moving arm carry out nucleic acid extraction, and driving sample goes out sample track and send sample to storage after the completion of extracting Area drives handgrip, the mechanical arm of detection device to be detected later.
Controller control specimen needle 14, reagent needle and bar magnet do not conflict, that is, can also mention in nucleic acid extraction to be other Take plate reagent adding and sample again.
Embodiment 2
As shown in figures 1-8
A method of the nucleic acid extraction and detection of the full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions, including Following steps:
Specimen holder to be measured send to injection port 4 to wait and be loaded through sample introduction track, and identifies sample letter through barcode reader 13 Breath, the sample if necessary to mix then are loaded again after rotation mixes in mark product evenly mixing device 6, send after being loaded through going out sample track Sample storage area 12 after to detection;
It extracts reagent and pushes to reagent track 16 from agent bin, and add other additional now with reagent;
Sample needle 14 is drawn sample and is added in extraction reagent;
Reagent track 16 will extract reagent and be pushed to nucleic acid extraction position 21 and fix;
The bar magnet 19 that corresponding core extracts position 21 completes nucleic acid extraction work in corresponding position, pushes to reagent after having extracted Storehouse is detected, and closes extraction door;
Extract reagent push to detection device sample-adding position, detection reagent storehouse will test reagent (including reaction enzymes, buffer, DNTP, primer etc.) push to reagent track 16, specimen needle 14 is drawn the nucleic acid extracted and is added in detection reagent, extract reagent by Track pushes to sample storage warehouse 29 after inspection;
Reagent transparent plastic after sample-adding is covered and is sent into detection device reaction tray 25 and detects;
Sample to be examined reacts under constant temperature conditions (65 DEG C), and carries out fluorescence detection by CCD, and reaction tray is centrifugation Formula, it can be ensured that reaction system mixes.Reaction tray is resetted per minute and carries out first order fluorescence detection;
Sample is pushed out reaction tray 25 by sample exhaust port 26 after inspection after having detected.
In the description of this specification, the description of reference term " one embodiment ", " example ", " specific example " etc. means Particular features, structures, materials, or characteristics described in conjunction with this embodiment or example are contained at least one of the utility model In embodiment or example.In the present specification, schematic expression of the above terms be not necessarily referring to identical embodiment or Example.Moreover, particular features, structures, materials, or characteristics described can be in any one or more embodiment or examples In can be combined in any suitable manner.
The preferred embodiment in the utility model disclosed above is only intended to help to illustrate the utility model.Preferred embodiment is simultaneously There is no the details that detailed descriptionthe is all, also not limiting the utility model is only the specific embodiment.Obviously, according to this theory The content of bright book can make many modifications and variations.These embodiments are chosen and specifically described to this specification, is in order to preferably The principles of the present invention and practical application are explained, so that skilled artisan be enable to better understand and utilize this Utility model.The utility model is limited only by the claims and their full scope and equivalents.
In the description of this specification, the description of reference term " one embodiment ", " example ", " specific example " etc. means Particular features, structures, materials, or characteristics described in conjunction with this embodiment or example are contained at least one of the utility model In embodiment or example.In the present specification, schematic expression of the above terms be not necessarily referring to identical embodiment or Example.Moreover, particular features, structures, materials, or characteristics described can be in any one or more embodiment or examples In can be combined in any suitable manner.
The preferred embodiment in the utility model disclosed above is only intended to help to illustrate the utility model.Preferred embodiment is simultaneously There is no the details that detailed descriptionthe is all, also not limiting the utility model is only the specific embodiment.Obviously, according to this theory The content of bright book can make many modifications and variations.These embodiments are chosen and specifically described to this specification, is in order to preferably The principles of the present invention and practical application are explained, so that skilled artisan be enable to better understand and utilize this Utility model.The utility model is limited only by the claims and their full scope and equivalents.

Claims (5)

1. a kind of full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions, it is characterised in that: including nucleic acid-extracting apparatus, Detection device and controller, nucleic acid-extracting apparatus are connected with detection device and are respectively equipped with gas-tight silo, detection device and extraction dress Set respectively with controller and connect to power supply;
The nucleic acid-extracting apparatus includes that sample sample introduction track, reagent track, nucleic acid extraction unit and sample go out sample track, described The injection port of sample sample introduction track connects specimen holder propeller, and rear side installs sample blending device, and connects reagent track, reagent Track two sides separately design by mechanical arm control reagent suction needle and sample suction needle, and be correspondingly arranged extract agent bin, TIP storehouses and waste holder, reagent track end connect each nucleic acid extraction unit, the nucleic acid extraction list by branching agent track It is reagent track among first, track two sides are nucleic acid extraction position, and the bar magnet of the upper layer installation specification arrangement of nucleic acid extraction unit is simultaneously It is controlled by bar magnet moving arm, nucleic acid extraction unit rear end connection sample goes out sample track, and sample track end is the mark after extracting out Product storage area;
The detection device includes the image sensor apparatus of reaction tray and charge-coupled device, is equipped in the reaction tray multiple Reaction cup, and design sample outlet and detection reagent sample introduction rail, sample exhaust port and detection reagent sample introduction track and reaction tray Mechanical gripper is designed in connecting pin, and sample exhaust port connects sample waste storage storehouse, installation sample envelope on detection reagent sample introduction track Lid device designs liquid injection port and temperature detection mouth in reaction tray.
2. a kind of full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions as described in claim 1, it is characterised in that: The nucleic acid-extracting apparatus upper end connects TIP loading channels and reagent storage storehouse, the nucleic acid extraction unit and detection unit Ultraviolet lamp is installed.
3. a kind of full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions as described in claim 1, it is characterised in that: TIP storehouses, agent bin and sample suction needle and propeller by mechanical arm control in the detection device;
Exhaust equipment is equipped on rear side of nucleic acid-extracting apparatus and nucleic acid detection apparatus;
The agent bin is refrigeration warehouse.
4. a kind of full-automatic nucleic acid amplification assays instrument as claimed in any one of claims 1-3 based on isothermal amplification reactions, It is characterized by: the full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions further includes control system, the control System includes controller, and the controller includes detection unit, control unit, execution unit and display unit, detection unit packet Barcode reader, the reaction tray temperature measuring probe of sample introduction track are included, control unit and display unit are to be mounted on controller Main control chip and display, execution unit include the control mechanical arm of reagent needle and specimen needle, sample sample introduction track, reagent rail Road, sample go out sample track, the handgrip of bar magnet moving arm and detection device, mechanical arm;Barcode reader detection signal feeds back to control Sample is sent to nucleic acid extraction unit, and drive control by the sample sample introduction track of unit processed, control unit driving executive component Mechanical arm, reagent track and bar magnet moving arm carry out nucleic acid extraction, after the completion of extracting driving sample go out sample track by mark product send to Storage area drives handgrip, the mechanical arm of detection device to be detected later.
5. the full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions as claimed in claim 4, it is characterised in that: described Controller control specimen needle, reagent needle and bar magnet do not conflict, that is, can also add an examination of again for other extraction positions in nucleic acid extraction Agent and sample.
CN201821502049.7U 2018-09-14 2018-09-14 A kind of full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions Expired - Fee Related CN209193976U (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108949548A (en) * 2018-09-14 2018-12-07 昆明医科大学第附属医院 A kind of full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108949548A (en) * 2018-09-14 2018-12-07 昆明医科大学第附属医院 A kind of full-automatic nucleic acid amplification assays instrument based on isothermal amplification reactions

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