CN112501006A - Automatic virus processing method - Google Patents
Automatic virus processing method Download PDFInfo
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- CN112501006A CN112501006A CN202011605144.1A CN202011605144A CN112501006A CN 112501006 A CN112501006 A CN 112501006A CN 202011605144 A CN202011605144 A CN 202011605144A CN 112501006 A CN112501006 A CN 112501006A
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- virus
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- nucleic acid
- sample
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- 241000700605 Viruses Species 0.000 title claims abstract description 48
- 238000003672 processing method Methods 0.000 title claims description 16
- 238000010276 construction Methods 0.000 claims abstract description 18
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 18
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 18
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 18
- 238000000034 method Methods 0.000 claims abstract description 16
- 230000002779 inactivation Effects 0.000 claims abstract description 10
- 238000005138 cryopreservation Methods 0.000 claims abstract description 9
- 230000003321 amplification Effects 0.000 claims abstract description 5
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 5
- 238000002360 preparation method Methods 0.000 claims abstract description 5
- 239000000523 sample Substances 0.000 claims description 35
- 238000007710 freezing Methods 0.000 claims description 19
- 230000008014 freezing Effects 0.000 claims description 19
- 230000008569 process Effects 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 11
- 239000002184 metal Substances 0.000 claims description 9
- 238000012545 processing Methods 0.000 claims description 9
- 239000011148 porous material Substances 0.000 claims description 7
- 239000011259 mixed solution Substances 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 5
- 238000000746 purification Methods 0.000 claims description 5
- 230000005540 biological transmission Effects 0.000 claims description 4
- 230000009471 action Effects 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 3
- 239000012488 sample solution Substances 0.000 claims description 3
- 238000001514 detection method Methods 0.000 abstract description 15
- 238000001821 nucleic acid purification Methods 0.000 abstract description 5
- 238000005070 sampling Methods 0.000 abstract description 3
- 238000003752 polymerase chain reaction Methods 0.000 abstract 2
- 239000000047 product Substances 0.000 description 4
- 238000012546 transfer Methods 0.000 description 4
- 230000010355 oscillation Effects 0.000 description 3
- 238000005034 decoration Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000012521 purified sample Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000000105 evaporative light scattering detection Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012946 outsourcing Methods 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/48—Automatic or computerized control
Abstract
The invention relates to the technical field of automation equipment, and discloses a virus automatic treatment method which is realized by applying a virus automatic treatment system, wherein the virus automatic treatment system comprises a biological safety cabinet, a virus pretreatment unit, a nucleic acid purifier, a system construction unit, a PCR (polymerase chain reaction) instrument and a control unit, and the steps of preparation work, pretreatment cryopreservation, inactivation, nucleic acid purification, system construction, amplification detection and the like are realized through the control unit. The invention is matched with virus automatic detection to complete the automation of a system operation from virus sampling to detection results.
Description
Technical Field
The invention relates to the technical field of laboratory automation, in particular to a virus automatic processing method.
Background
The nucleic acid detection process, from sampling, liquid separation, purification, amplification to the final detection result, all needs manual work, and is complex and long in time consumption. There is also certain safety risk in the detection process, and enough personnel and related auxiliary equipment need to be equipped to meet the national demand for realizing rapid and mass detection in a specific area, so that the cost and the detection time are greatly increased, and the accuracy of the detection result is influenced to a certain extent.
Disclosure of Invention
The present invention is directed to solve the above problems, and an object of the present invention is to provide an automatic virus processing method, which is used in conjunction with automatic virus detection to complete automation of system operations from virus sampling to detection results.
The technical scheme adopted by the invention is as follows:
a virus automatic processing method is realized by a virus automatic processing system, the virus automatic processing system comprises a biological safety cabinet, a virus pretreatment unit, a nucleic acid purifier, a system construction unit, a PCR instrument and a control unit, and is characterized in that the following steps are realized by the control unit:
(1) arranging samples and consumables in a virus pretreatment unit in the biosafety cabinet; arranging consumables in a nucleic acid purifier and a PCR instrument; ensuring the safety of each device and preparing for work;
(2) oscillating a sample bottle in the virus pretreatment unit;
(3) carrying out uncovering operation on the sample bottle to be processed and the three freezing tubes;
(4) quantitatively transferring the sample solution in the sample bottle to three freezing tubes;
(5) closing the three freezing pipes, returning two freezing pipes, and sending one freezing pipe to a metal bath;
(6) repeating the steps (3) to (5), and completely completing pipetting for the preset cryopreservation tubes;
(7) after the cryopreservation tube finishes metal bath inactivation, opening the cover, and subpackaging the inactivated sample to a pore plate;
(8) outputting the product through a pore plate, and purifying the product by a nucleic acid purifier;
(9) after purification is finished, sending the mixture to a PCR system for construction, and completing the preparation of mixed liquor after the PCR system is constructed;
(10) constructing a PCR system, and configuring the mixed solution and the inactivated sample to generate a PCR reaction plate;
(11) centrifuging the PCR reaction plate by a centrifuge;
(12) and (4) conveying the PCR reaction plate to a PCR instrument for amplification.
Further, the samples in the step (1) are arranged on a vibration device by the sample bottle array, and the vibration of the sample bottles is realized by the vibration device in the step (2).
Further, the steps (3) to (5) are completed through a turntable unit, and the turntable unit simultaneously processes the actions of uncovering, pipetting and covering the three cryopreservation tubes.
Further, the pipetting processes in the steps (4) and (7) change the pipetting head when one sample is finished.
Further, the metal bath inactivation temperature in the step (7) is 56 ℃ for 30 minutes.
Further, in the step (9), the mixed solution in the PCR system construction is completed synchronously at the time of the operation of the step (8).
Further, in the step (11), two PCR reaction plates are subjected to a set of centrifugation.
Further, the container transmission among the biological safety cabinet, the virus pretreatment unit, the nucleic acid purifier, the system construction unit and the PCR instrument is completed through robot operation.
Further, the control unit comprises an upper computer and a lower computer, the virus pretreatment unit and the system construction unit are controlled by the lower computer, and the upper computer controls the biological safety cabinet, the nucleic acid purifier, the PCR instrument and the lower computer.
Further, the control of the biological safety cabinet, the nucleic acid purifier and the PCR instrument is completed by operating working buttons of the robot.
The invention has the beneficial effects that:
(1) the whole process is automatically realized from sample collection to detection results;
(2) each sample forms a detection chain, and data can be tracked;
(3) the processing flow is reasonable in configuration, and the detection result is accurate and reliable.
Drawings
FIG. 1 is a flow chart of the present invention.
Detailed Description
The following describes in detail a specific embodiment of the automated virus processing method according to the present invention with reference to the drawings.
Referring to fig. 1, the virus automatic processing method is realized by using a virus automatic processing system, the virus automatic processing system comprises a biological safety cabinet, a virus pretreatment unit, a nucleic acid purifier, a system construction unit, a PCR instrument and a control unit, and each unit and equipment respectively complete the specific functions of each stage. The biological safety cabinet is used for the safety guarantee function of virus pretreatment, the virus pretreatment unit finishes sample liquid separation cryopreservation and inactivation, the nucleic acid purifier performs nucleic acid purification, the systemic system construction unit performs mixed liquid treatment on the purified sample, and the purified sample is amplified by the PCR instrument to finish detection. The intelligent operation of the robot is provided, all units and equipment are connected in series to form an organic whole, and the control unit completes the implementation of the whole process.
The method comprises the following specific steps:
(1) carry out sample and consumptive material to the virus pretreatment unit in the biosafety cabinet and arrange, arrange on vibrating device by sample bottle array after the sample is gathered in advance. The arrangement of consumables is carried out in nucleic acid purification appearance and PCR appearance, and the biosafety cabinet adopts the 1379 model of the seemer feishel brand, and the nucleic acid purification appearance is the MagNA Pure96 model of luo shi brand, and the PCR appearance is the LightCycler480 II model of luo shi brand. Corresponding consumable items can be configured according to specific products, and the consumable items are strictly executed according to product specifications. The safety of each device is ensured and other corresponding work preparations are made.
(2) And oscillating the sample bottle in the virus pretreatment unit, wherein the oscillation process is finished on an oscillation device, and the oscillation frequency and time are set according to the source and the type of the collected sample so as to fully mix and dissolve the effective substances in the sample in the solution of the sample bottle.
(3) And (3) carrying out uncovering operation on the sample bottle to be processed and the three freezing tubes, wherein the freezing tubes are pre-arranged in the virus pretreatment unit, and the uncovering action of the three freezing tubes at each time is completed through the mechanical arm and the arranged turntable unit.
(4) Quantitatively transferring the sample solution in the sample bottle to the three freezing tubes, uncovering the turntable unit at one station, and transferring liquid at one station. And the pipetting head is replaced according to the flow requirement in the pipetting process.
(5) After the pipetting is finished, the three freezing tubes are sequentially covered, the two freezing tubes are returned through the mechanical arm, and one freezing tube is conveyed to the metal bath.
(6) And (5) repeating the steps (3) to (5) and completely completing the pipetting on the preset freezing storage tubes. Before the sample bottle is uncovered during pipetting at every time, the sample bottle is scanned, and data are transmitted to a background database. And the sample bottle after liquid transfer is put back to the original position after the cover is closed.
(7) After the cryopreservation tube finishes metal bath inactivation, opening the cover, and subpackaging the inactivated sample to a pore plate; inactivation procedure the metal bath inactivation temperature was 56 ℃ for 30 minutes. And meanwhile, the pipetting head is replaced according to the flow requirement in the pipetting process.
(8) After the inactivation is completed, the inactivated cryopreservation tube is uncapped on the turntable unit, and the inactivated sample is transferred to the pore plate for output and then sent to the nucleic acid purifier for purification. Meanwhile, the PCR system construction is carried out in advance to prepare the mixed solution.
(9) And after the purification is finished, sending the mixture to a PCR system for construction, and finishing the mixture preparation at the moment of the construction of the PCR system.
(10) And (3) constructing a PCR system, configuring the mixed solution and the inactivated sample to generate a PCR reaction plate, and packaging and outputting the reaction plate. The PCR reaction plate is filled with an inactivated sample and mixed liquid, liquid transfer is realized through a liquid injection and ejection gun, and a liquid transfer head is prepared to be replaced in the transfer process according to the principle that the samples are not crossed.
(11) And centrifuging the PCR reaction plate by a centrifuge to enable the liquid drops in the PCR reaction plate to process the lowest position of the pore plate, and realizing centrifugation of a double-plate group during centrifugal operation.
(12) And (5) conveying the PCR reaction plate to a PCR instrument for amplification to finish detection. After completion, each consumable and the abandoned part are recovered, and the data chain data supplement is complete.
The container transmission among the biological safety cabinet, the virus pretreatment unit, the nucleic acid purifier, the system construction unit and the PCR instrument is completed through robot operation. The lower computer of each device is connected to the upper computer of the console for control. Because biosafety cabinet, nucleic acid purification appearance, PCR appearance are current outsourcing equipment, can't realize system access through the next computer, use the robot to realize the instruction transmission to equipment through hard interface, through simulating manual operation, operate the control that realizes equipment to the button of equipment promptly.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A virus automatic processing method is realized by a virus automatic processing system, the virus automatic processing system comprises a biological safety cabinet, a virus pretreatment unit, a nucleic acid purifier, a system construction unit, a PCR instrument and a control unit, and is characterized in that: the following steps are realized by the control unit:
(1) arranging samples and consumables in a virus pretreatment unit in the biosafety cabinet; arranging consumables in a nucleic acid purifier and a PCR instrument; ensuring the safety of each device and preparing for work;
(2) oscillating a sample bottle in the virus pretreatment unit;
(3) carrying out uncovering operation on the sample bottle to be processed and the three freezing tubes;
(4) quantitatively transferring the sample solution in the sample bottle to three freezing tubes;
(5) closing the three freezing pipes, returning two freezing pipes, and sending one freezing pipe to a metal bath;
(6) repeating the steps (3) to (5), and completely completing pipetting for the preset cryopreservation tubes;
(7) after the cryopreservation tube finishes metal bath inactivation, opening the cover, and subpackaging the inactivated sample to a pore plate;
(8) outputting the product through a pore plate, and purifying the product by a nucleic acid purifier;
(9) after purification is finished, sending the mixture to a PCR system for construction, and completing the preparation of mixed liquor after the PCR system is constructed;
(10) constructing a PCR system, and configuring the mixed solution and the inactivated sample to generate a PCR reaction plate;
(11) centrifuging the PCR reaction plate by a centrifuge;
(12) and (4) conveying the PCR reaction plate to a PCR instrument for amplification.
2. The automated virus processing method according to claim 1, wherein: the samples in the step (1) are arranged on the oscillating device through the sample bottle array, and the oscillating of the sample bottles is realized through the oscillating device in the step (2).
3. The automated virus processing method according to claim 1, wherein: and (5) completing the steps (3) to (5) through a turntable unit, and simultaneously processing the actions of opening the cover of the three freezing tubes, transferring liquid and closing the cover.
4. The automated virus processing method according to claim 1, wherein: and (4) in the pipetting processes in the steps (4) and (7), the pipetting head is replaced when one sample is finished.
5. The automated virus processing method according to claim 1, wherein: the metal bath inactivation temperature in the step (7) is 56 ℃ for 30 minutes.
6. The automated virus processing method according to claim 1, wherein: in the step (9), the mixed solution in the PCR system construction is synchronously completed during the operation of the step (8).
7. The automated virus processing method according to claim 1, wherein: in the step (11), two PCR reaction plates complete a set of centrifugation operation.
8. The method for automated virus processing according to any one of claims 1 to 7, wherein: the container transmission among the biological safety cabinet, the virus pretreatment unit, the nucleic acid purifier, the system construction unit and the PCR instrument is completed through robot operation.
9. The automated virus processing method according to claim 8, wherein: the control unit comprises an upper computer and a lower computer, the virus pretreatment unit and the system construction unit are controlled by the lower computer, and the upper computer controls the biological safety cabinet, the nucleic acid purifier, the PCR instrument and the lower computer.
10. The automated virus processing method according to claim 8, wherein: the control of the biological safety cabinet, the nucleic acid purifier and the PCR instrument is completed by operating working buttons of the robot.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011605144.1A CN112501006A (en) | 2020-12-30 | 2020-12-30 | Automatic virus processing method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011605144.1A CN112501006A (en) | 2020-12-30 | 2020-12-30 | Automatic virus processing method |
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| CN112501006A true CN112501006A (en) | 2021-03-16 |
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN210481395U (en) * | 2020-04-04 | 2020-05-08 | 博奥生物集团有限公司 | High-throughput full-automatic nucleic acid detection system |
| CN111217309A (en) * | 2020-04-04 | 2020-06-02 | 倍仪昇智能科技(苏州)有限公司 | Full-automatic liquid-separating and capping system and liquid-separating and capping method |
| CN111269808A (en) * | 2020-02-17 | 2020-06-12 | 英诺维尔智能科技（苏州）有限公司 | Automatic safety detection method for high-pollution high-infection-risk biological samples |
| CN111286444A (en) * | 2020-02-27 | 2020-06-16 | 中国科学院上海技术物理研究所 | High-sensitivity virus rapid detector |
| CN111778157A (en) * | 2020-08-13 | 2020-10-16 | 上海理工大学 | High-protection-level automatic flexible pathogenic nucleic acid detection system and method |
| US20200363299A1 (en) * | 2016-11-23 | 2020-11-19 | Hangzhou Gene-Meta Medical Device Co., Ltd. | Apparatus for automating pretreatment of nucleic acid detection |
| CN112080421A (en) * | 2020-08-28 | 2020-12-15 | 中国科学院苏州生物医学工程技术研究所 | Ultrahigh-flux full-automatic pathogen nucleic acid detection system and method |
| CN112094737A (en) * | 2020-09-22 | 2020-12-18 | 山东省科学院能源研究所 | Full-automatic virus sample pretreatment device and control method thereof |
-
2020
- 2020-12-30 CN CN202011605144.1A patent/CN112501006A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20200363299A1 (en) * | 2016-11-23 | 2020-11-19 | Hangzhou Gene-Meta Medical Device Co., Ltd. | Apparatus for automating pretreatment of nucleic acid detection |
| CN111269808A (en) * | 2020-02-17 | 2020-06-12 | 英诺维尔智能科技（苏州）有限公司 | Automatic safety detection method for high-pollution high-infection-risk biological samples |
| CN111286444A (en) * | 2020-02-27 | 2020-06-16 | 中国科学院上海技术物理研究所 | High-sensitivity virus rapid detector |
| CN210481395U (en) * | 2020-04-04 | 2020-05-08 | 博奥生物集团有限公司 | High-throughput full-automatic nucleic acid detection system |
| CN111217309A (en) * | 2020-04-04 | 2020-06-02 | 倍仪昇智能科技(苏州)有限公司 | Full-automatic liquid-separating and capping system and liquid-separating and capping method |
| CN111778157A (en) * | 2020-08-13 | 2020-10-16 | 上海理工大学 | High-protection-level automatic flexible pathogenic nucleic acid detection system and method |
| CN112080421A (en) * | 2020-08-28 | 2020-12-15 | 中国科学院苏州生物医学工程技术研究所 | Ultrahigh-flux full-automatic pathogen nucleic acid detection system and method |
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Application publication date: 20210316 |
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