CN116757233A - Traceable automatic nucleic acid detection system and method - Google Patents
Traceable automatic nucleic acid detection system and method Download PDFInfo
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- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 191
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- 239000012807 PCR reagent Substances 0.000 claims abstract description 115
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- 238000004458 analytical method Methods 0.000 claims description 17
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Abstract
The invention discloses a traceable automatic nucleic acid detection system and a method, wherein the automatic nucleic acid detection system comprises the following components: the sample information acquisition module is used for realizing sample information input and uploading of a sample to be detected; the nucleic acid extraction module is used for realizing nucleic acid extraction of a sample to be detected; the PCR system preparation module is used for adding the extracted product nucleic acid into a PCR reagent system to complete the construction of a PCR reaction system; the nucleic acid detection module is used for amplifying the nucleic acid to be detected and completing detection; and the information processing device is used for completing information recording, processing and transmitting at each stage in the sample processing process. The system and/or the method of the invention improve the automation degree of the nucleic acid detection flow, the rapidness and the reliability of sample tracing.
Description
Technical Field
The invention belongs to the technical field of nucleic acid detection, and particularly relates to a traceable automatic nucleic acid detection system and method.
Background
In the process of extracting and detecting nucleic acid in the prior art, the following defects exist: (1) in a conventional nucleic acid detection laboratory, except for nucleic acid extraction and nucleic acid detection automation equipment, the steps of sample pretreatment, PCR reagent system, PCR reaction system construction and the like are related, and are usually completed manually by an experimenter, so that the automation degree in the sample treatment process is low, a large amount of manpower is consumed, the efficiency is low, and errors are easy to occur; (2) before the detection result is not received, the sample tube is usually concentrated in an idle place by an experimenter, sample management is not standard, and when the sample with abnormal result is required to be rechecked, the experimenter is required to search and check one by one, so that the time and the labor are consumed, and errors are easy to occur.
Disclosure of Invention
The invention aims at: in order to overcome the problems in the prior art, a traceable automatic nucleic acid detection system and a traceable automatic nucleic acid detection method are disclosed, and the automation degree of a nucleic acid detection process, and the rapidness and the reliability of sample traceability are improved through the system and/or the method.
On the one hand, the aim of the invention is achieved by the following technical scheme:
a traceable automated nucleic acid detection system, the automated nucleic acid detection system comprising:
The sample information acquisition module is used for realizing sample information input and uploading of a sample to be detected;
the nucleic acid extraction module is used for realizing nucleic acid extraction of a sample to be detected;
the PCR system preparation module is used for adding the extracted product nucleic acid into a PCR reagent system to complete the construction of a PCR reaction system;
the nucleic acid detection module is used for amplifying the nucleic acid to be detected and completing detection;
and the information processing device is used for completing information recording, processing and transmitting at each stage in the sample processing process.
According to a preferred embodiment, the sample information acquisition module comprises a sample tube, a sample rack and a first scanning device; the sample tube is placed on the sample frame, and identification codes are respectively arranged on the sample tube and the sample frame; the sample rack is also provided with a position sensor, and position information acquisition and uploading of each sample tube on the sample rack are completed through the position sensor; the first scanning device is used for collecting and uploading identification codes of the sample tube and the sample frame; and the information processing device is configured to realize that the sample tube identification code information corresponds to the sample frame identification code information and complete the correspondence of the sample tube identification code information and the position information of the sample tube.
According to a preferred embodiment, the nucleic acid extraction module comprises a sample pre-processor for performing an automatic transfer of the sample to be tested and a nucleic acid extraction instrument; the nucleic acid extractor is used for extracting the nucleic acid of the sample to be detected in the sample pretreatment apparatus.
According to a preferred embodiment, the sample pretreatment apparatus comprises a plurality of deep well plates with identification codes and a second scanning device, and the nucleic acid extraction apparatus further comprises a third scanning device; the deep hole plate is used for sub-packaging the transferred sample to be tested, and the second scanning device is used for completing collection and uploading of identification codes of the sample tube, the sample rack and the deep hole plate; the identification code of the deep hole plate records sample information of each sample to be detected which is grasped by the information processing device and an experimental program of the nucleic acid extractor; the nucleic acid extraction instrument completes nucleic acid extraction based on an experimental procedure obtained by scanning the identification code of the deep well plate by the third scanning device.
According to a preferred embodiment, the PCR system preparation module comprises a PCR reagent split charging instrument and a PCR reaction system constructing instrument, wherein the PCR reagent split charging instrument obtains a PCR reagent system preparation program through an information processing device to complete the preparation of a PCR reagent system; the PCR reaction system construction instrument is used for adding the extracted product nucleic acid into a PCR reagent system to complete the construction of the PCR reaction system.
According to a preferred embodiment, the PCR reagent dispensing apparatus comprises a fourth scanning device and a plurality of PCR reagent plates; the PCR reagent plate is used for realizing premixing and split charging of the PCR reagents, and the identification code of the PCR reagent plate records the experimental program of the PCR reaction system constructor; the fourth scanning device is used for scanning the identification code of the PCR reagent plate; the PCR reaction system construction instrument further comprises a fifth scanning device, wherein the fifth scanning device is used for scanning an identification code of a deep pore plate filled with extracted nucleic acid and an identification code of a PCR reagent plate, and the PCR reagent plate (301) filled with a sample for completing the construction of a PCR reaction system is used as a reaction system PCR plate (302); the information processing device transmits sample information recorded in the identification code of the deep hole plate filled with the nucleic acid to the identification code of the reaction system PCR plate. The nucleic acid extracted from the sample is added to the PCR reagent plate to form a reaction system PCR plate, that is, the reaction system PCR plate is identical to the PCR reagent plate, but the contained solution is different.
According to a preferred embodiment, the PCR reagent dispensing device further comprises: a reagent and premixing zone, a Tip zone, a waste zone and a PCR reagent loading zone; the reagent and the premixing area are used for placing reagents required by the PCR reagent system and premixing the reagents; the Tip region is used for loading samplers with different specifications; the waste area is used for accommodating an automatically unloaded sampler; the PCR reagent loading area is used for placing a PCR reagent plate containing a PCR reagent system.
According to a preferred embodiment, the PCR reaction system builder further comprises: the system establishment area, the Tip area, the product extraction area, the waste area and the product sample reserving area; the system establishment area is used for placing a PCR reagent plate filled with a PCR reagent, and the Tip area is used for loading samplers with different specifications; the extraction product region is used for loading a deep hole plate filled with extraction product nucleic acid; the waste area is used for accommodating an automatically unloaded sampler; the product retention zone is used to retain the remaining extracted product.
According to a preferred embodiment, the nucleic acid detecting module comprises a nucleic acid detector, a computer end and a sixth scanning device, wherein the computer end is used for analyzing and sorting detection data of the nucleic acid detector and uploading analysis results to the information processing device; the sixth scanning device is used for scanning the identification codes of the PCR plates of the reaction system, and the information processing device is used for corresponding the sample information to the analysis result of the detection data.
According to a preferred embodiment, the automated nucleic acid detection system further comprises a sample receiving module for receiving, managing sample tubes for which sample transfer has been completed.
According to a preferred embodiment, the sample storage module comprises a plurality of sample cabinets, and each sample cabinet is internally provided with a frame position with a position code for placing a sample frame; each sample cabinet further comprises a seventh scanning device and a sample cabinet management unit, the sample detection state on the sample rack in the sample cabinet and the visual display of the position of the sample rack are completed through the sample cabinet management unit, and the seventh scanning device is used for scanning the identification code of the sample rack, the serial number of the sample cabinet and the position code of the rack position; the identification code of the sample rack, the serial number of the sample cabinet and the position code of the rack position are respectively corresponding through the information processing device; and feeding back the analysis result of the nucleic acid detection data to the sample cabinet management unit through the information processing device.
In another aspect, the invention also discloses:
a traceable automated nucleic acid detection method, the automated nucleic acid detection method comprising:
step 1: sample information input, namely inputting sample information, wherein the sample information comprises identification code information of a sample tube, a sample frame and position information of the sample tube on the sample frame; step 2: a pre-treatment step of the sample to be measured, completing automatic transfer of the sample to be measured; step 3: extracting nucleic acid of a sample to be detected; step 4: preparing a PCR system; adding the extracted product nucleic acid into a PCR reagent system to complete the construction of a PCR reaction system; step 5: a nucleic acid detection step of amplifying and detecting the nucleic acid to be detected; step 6: and a sample tube storage management step of storing and managing the sample tubes with the samples transferred, and displaying the analysis results of the nucleic acid detection data of each sample tube.
According to a preferred embodiment, the sample information entering step of step 1 comprises: the first scanning device scans the identification code information on the sample tube and the sample frame and uploads the identification code information to the information processing device, and meanwhile, the position sensor on the sample frame uploads the recorded position information of the sample tube to the information processing device, and the information processing device corresponds the identification code information on the sample tube, the identification code information on the sample frame and the position information of the sample tube on the sample frame; the information processing device respectively sends a sample automatic transfer experiment program, a PCR reagent system preparation program and a nucleic acid detection experiment program to a sample pretreatment instrument, a PCR reagent split charging instrument and a nucleic acid detector according to the number of the recorded identification codes of the sample tubes.
According to a preferred embodiment, the pre-processing step of the sample to be tested of step 2 comprises: the sample pretreatment device is used for automatically transferring samples to be tested in the sample tube into a deep hole plate with identification codes based on a sample automatic transfer experiment program, the second scanning device scans the identification code information of the sample tube, the identification code information of the sample frame and the identification codes of the deep hole plate, and the information processing device transmits the sample information into the identification codes of the deep hole plate to realize the correspondence between the identification codes of the deep hole plate and the sample information; meanwhile, the nucleic acid extraction experimental program is transferred to the identification code of the deep well plate through the information processing device.
According to a preferred embodiment, the nucleic acid extraction step of the sample to be tested of step 3 comprises: the third scanning device scans the identification code of the deep hole plate, the nucleic acid extractor acquires and runs a nucleic acid extraction experiment program through the identification code of the deep hole plate, extracts nucleic acid from the sample to be detected transferred into the deep hole plate, and simultaneously grabs sample information recorded in the identification code of the deep hole plate.
According to a preferred embodiment, the PCR system formulation step of step 4 comprises: the PCR reagent split charging instrument obtains a PCR reagent system preparation program through the information processing device to complete the preparation of the PCR reagent system; meanwhile, the fourth scanning device scans the identification code of the PCR reagent plate filled with the PCR reagent system and uploads the identification code to the information processing device, and the information processing device records the experimental program of the construction of the PCR reaction system into the identification code of the PCR reagent plate;
Then, a fifth scanning device scans the identification code of the PCR reagent plate and the identification code of the deep hole plate containing the extracted product nucleic acid, and uploads the identification code to the information processing device, a PCR reaction system construction instrument acquires and operates a PCR reaction system construction experiment program to complete the preparation of a PCR reaction system, and a sample constructed by the PCR reaction system is contained in the reaction system PCR plate; the information processing device transmits sample information corresponding to the identification code of the deep pore plate containing the nucleic acid to the identification code of the PCR plate of the reaction system.
According to a preferred embodiment, the nucleic acid detection step of step 5 comprises: the nucleic acid detector acquires and runs a nucleic acid detection experiment program through the information processing device, completes nucleic acid detection of a sample to be detected, outputs detection data to a computer end, analyzes and sorts the detection data by the computer end, and then uploads a data analysis result to the information processing device; and the sixth scanning device scans the identification code of the PCR plate of the reaction system, and uploads the identification code to the information processing device, and the information processing device corresponds the data analysis result to the sample information.
According to a preferred embodiment, the sample tube storage management step of step 6 includes: a seventh scanning device scans the identification code information of the sample rack carrying the sample tubes, the serial numbers of the sample cabinets and the position codes of the sample rack positions, and places the sample rack carrying the sample tubes on the positions in the sample cabinets; the information processing device feeds back the sample detection data analysis result to the sample cabinet management unit, and the sample cabinet management unit performs visual display on the position of the sample frame and the detection data analysis result of the sample on the sample frame.
The foregoing inventive concepts and various further alternatives thereof may be freely combined to form multiple concepts, all of which are contemplated and claimed herein. Various combinations will be apparent to those skilled in the art from a review of the present disclosure, and are not intended to be exhaustive or all of the present disclosure.
The invention has the beneficial effects that:
the traceable automatic nucleic acid detection system and/or method provided by the invention mainly have the following technical advantages:
(1) The sample information acquisition module, the nucleic acid extraction module, the PCR system preparation module, the nucleic acid detection module, the sample storage module and the information processing device are cooperatively used, so that the automatic processing of each link of sample processing is realized, the full-flow transmission and monitoring of sample information are realized, and the reliability of sample information verification is improved; and moreover, the automatic transmission and identification of experimental procedures in each link are realized, and the time consumed by manually setting the experimental procedures is shortened.
(2) By arranging the sample pretreatment instrument and the PCR system preparation module, the automatic treatment of sample transfer, PCR reagent system construction and PCR reaction system construction is realized, a great deal of manpower is saved, the preparation speed of the PCR reaction system is improved, and the manual operation errors are reduced.
(3) Through setting up sample and accomodating the module, realized waiting the visual standard management of the sample of testing result, make things convenient for the experimenter to seek the sample that needs the recheck, improved sample traceability efficiency, the experimenter of being convenient for has been accomplished the management of the sample that detects.
(4) By means of the technical scheme, the processing speed of the sample to be detected is increased, the daily sample detection amount is further increased, and the sample processing pressure of large-scale nucleic acid screening is reduced.
Drawings
FIG. 1 is a schematic diagram of the composition of a traceable automated nucleic acid detection system in accordance with an embodiment of the present invention;
FIG. 2 is a schematic diagram of a sample information acquisition module according to an embodiment of the present invention;
FIG. 3 is a schematic diagram showing the constitution of a nucleic acid extraction module according to an embodiment of the present invention;
FIG. 4 is a schematic diagram of the structure of a PCR system preparation module according to an embodiment of the present invention;
FIG. 5 is a schematic diagram showing the constitution of a nucleic acid detecting module according to an embodiment of the present invention;
FIG. 6 is a schematic view of a sample receiving module according to an embodiment of the present invention;
FIG. 7 is a diagram of sample processing in an embodiment of the invention;
FIG. 8 is a schematic diagram showing the transfer of the position of samples in a deep well plate and a PCR plate according to an embodiment of the present invention.
The device comprises a 1-sample information acquisition module, a 101-sample tube, a 102-sample frame, a 103-first scanning device, a 2-nucleic acid extraction module, a 20-sample pretreatment instrument, a 21-nucleic acid extraction instrument, a 201-first deep pore plate, a 202-second deep pore plate, a 203-third deep pore plate, a 204-fourth deep pore plate, a 205-fifth deep pore plate, a 206-sixth deep pore plate, a 2011-second scanning device, a 2111-third scanning device, a 3-PCR system preparation module, a 30-PCR reagent split charging instrument, a 31-PCR reaction system construction instrument, a 300-fourth scanning device, a 310-fifth scanning device, a 301-PCR reagent plate, a 302-reaction system PCR plate, a 4-nucleic acid detection module, a 40-nucleic acid detection instrument, a 400-computer terminal, a 401-sixth scanning device, a 5-sample storage module, a 501-seventh scanning device, a 502-sample cabinet management unit, a 6-information processing device, a sample position before 13-extraction, and a 14-extraction product nucleic acid position.
Detailed Description
In order to more clearly illustrate the embodiments of the application or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the application, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Referring to fig. 1-8, a traceable automatic nucleic acid detection system in an embodiment of the application includes a sample information acquisition module 1, a nucleic acid extraction module 2, a PCR system preparation module 3, a nucleic acid detection module 4, a sample storage module 5, and an information processing device 6.
The sample information acquisition module 1 is used for inputting sample information and uploading the sample information to the information processing device 6. The sample information acquisition module 1 specifically comprises a sample tube 101 with an identification code and a sample frame 102 with the identification code, wherein the sample frame 102 is used for bearing the sample tube 101, and a position sensor is arranged on the sample frame 102 and used for recording position information of the sample tube 101 on the sample frame. The sample information includes sample tube 101 identification code information, sample holder 102 identification code information, and position information of the sample tube 101 on the sample holder 102, and the sample tube 101 identification code information, the sample holder 102 identification code information, and the position information of the sample tube 101 on the sample holder 102 are correlated by the information processing device 6.
The nucleic acid extraction module 2 is used for extracting nucleic acid from a sample to be detected, and specifically comprises a sample pretreatment instrument 20 and a nucleic acid extraction instrument 21; the sample pretreatment apparatus 20 is used for automatic transfer of the sample to be tested, and the transferred sample to be tested enters the nucleic acid extraction apparatus 21 for nucleic acid extraction.
In the present embodiment, the sample preprocessing instrument 20 receives the sample tube 101 to be measured carried on the sample rack 102 from the sample information acquisition module 1, and automatically transfers the sample to be measured into each of the deep well plates 201 to 206; the sample pretreatment apparatus 20 can be a fully automatic sample treatment system (model: geneMix Pro) developed by sienna technology limited company, and the sample pretreatment apparatus 20 can realize operations of automatically opening/closing a cover of a sample, transferring the sample, entering information of a deep hole plate, identifying a sample identification code, recycling a sample tube to a sample rack, loading the sample to a deep hole plate and the like, meanwhile, the sample pretreatment apparatus 20 is adapted to a 6 x 16t standard deep hole plate, and sub-packaging of 96 samples can be completed within 20 minutes.
In the present embodiment, the nucleic acid extraction instrument 21 receives the sample to be measured transferred from the sample pretreatment instrument 20 into each of the deep well plates 201 to 206, and performs nucleic acid extraction thereon. The nucleic acid extractor 21 may be a full-automatic nucleic acid extractor (model: gene Rotex 96) developed by Siam Tianlong technologies, inc., which can realize rapid and efficient preparation of a large amount of sample nucleic acids and is adapted to a 96-well deep well plate.
The PCR system preparation module 3 is used for preparing a PCR reaction system, and specifically comprises a PCR reagent sub-packaging instrument 30 and a PCR reaction system constructing instrument 31. The PCR reagent sub-packaging instrument 30 is used for constructing a PCR reagent system, and realizing PCR reagent premixing, PCR reagent transferring and PCR reagent sub-packaging; the PCR reaction system constructor 31 is used for adding the extracted product nucleic acid into a PCR reagent system to complete the construction of a reaction system before PCR detection.
In this embodiment, the PCR reagent sub-packaging apparatus 30 and the PCR reaction system construction apparatus 31 may specifically be full-automatic pipetting stations (model: PANA S401) developed by sienna technologies, ltd, and may be used to place required experimental consumables and reagents according to actual needs. Specifically, the PCR reagent split charging instrument 30 completes the construction of the PCR reagent system, and outputs the PCR reagent plate 301; the PCR reaction system constructing instrument 31 receives the PCR reagent plate 301 from the PCR reagent sub-packaging instrument 30 and the deep hole plates 201-206 filled with the extracted product nucleic acid after being extracted by the nucleic acid extracting instrument 21, and achieves the operation of adding the extracted product nucleic acid into the PCR reagent system, and a reaction system PCR plate 302 filled with the PCR reaction system is obtained, wherein the reaction system PCR plate 302 is consistent with the PCR reagent plate 301.
The nucleic acid detection module 4 is used for amplifying and detecting the extracted nucleic acid to obtain detection data. Specifically, the device comprises a nucleic acid detector 40 and a computer terminal 400 connected with the nucleic acid detector 40, which is used for analyzing and sorting detection data and uploading the data analysis result to the information processing device 6.
In this embodiment, the nucleic acid detector 40 is a real-time fluorescent quantitative PCR instrument, and may be a full-automatic medical PCR analysis system (model: gentier 96) developed by Siamiron technologies, inc. In this embodiment, the nucleic acid detector 40 receives the sample to be detected from the PCR reaction system construction instrument 31 to complete the PCR reaction system construction, and performs nucleic acid amplification and detection on the sample to be detected.
The sample storage module 5 is used for storing and managing the sample tube with the completed sample transfer. The sample cabinet comprises at least one sample cabinet 50 and a sample cabinet management unit 502, wherein a rack position with position codes is arranged in the sample cabinet 50 and used for placing a sample rack; the sample cabinet management unit 502 is used for visually displaying a sample detection state on a sample rack in the sample cabinet and the position of the sample rack; the sample cabinet management unit 502 is connected to the sample cabinet 50.
The information processing device 6 is in communication connection with the sample information acquisition module 1, the nucleic acid extraction module 2, the PCR system preparation module 3, the nucleic acid detection module 4 and the sample storage module 5 through a wired or wireless network, so that the collection and transmission of sample information of each link and the transmission of experimental programs of all instruments in the nucleic acid extraction module, the PCR system preparation module and the nucleic acid detection module are realized.
As shown in fig. 2 to 6, in this embodiment, the sample information collection module 1 further includes a first scanning device 103, where the first scanning device 103 is configured to sequentially scan the identification code on the sample tube 101 and the identification code on the sample holder 102, and upload the identification code information of the sample tube 101, the identification code information of the sample holder 102, and the position information of the sample tube 101 on the sample holder 102 to the information processing device 6, and the information processing device 6 corresponds the identification code information of the sample tube 101 and the identification code information of the sample holder 102, and the sample tube identification code information and the sample tube position information. In addition, the information processing device 6 transmits a sample automated transfer experiment program, a PCR reagent system preparation program, and a nucleic acid detection experiment program to the sample preprocessor 20, the PCR reagent sub-packaging apparatus 30, and the nucleic acid detector 40, respectively, according to the number of entered sample tube 101 identification codes.
The sample pretreatment apparatus 20 further comprises a second scanning device 2011 and deep-hole plates 201-206 with identification codes, wherein the second scanning device 2011 is used for sequentially scanning the identification codes on the sample tube 101, the identification codes on the sample rack 102 and the identification codes of the deep-hole plates 201-206; the deep hole plate 201-206 is used for sub-packaging the transferred sample to be tested, the identification code of the deep hole plate 201-206 captures the corresponding sample information contained in the deep hole plate 201-206 through the information processing device 6, and meanwhile, the information processing device 6 transmits the nucleic acid extraction experimental program to the identification code of the deep hole plate 201-206.
The nucleic acid extraction apparatus 21 further includes a third scanning device 2111, the third scanning device 2111 being used for scanning the identification code of the deep-hole plate 201-206 containing the extracted product nucleic acid, grasping sample information by the information processing device 6, and at the same time, the nucleic acid extraction apparatus 21 acquiring and running a nucleic acid extraction program; the deep-well plates 201-206 filled with the extracted product nucleic acids correspond to the deep-well plates 201-206 filled with the transferred samples to be tested.
The PCR reagent dispensing apparatus further includes a fourth scanning device 300 for scanning the identification code of the PCR reagent plate 301; the information processing device 6 records the PCR reaction system construction experiment program in the identification code of the PCR reagent plate 301.
The PCR reaction system constructor 31 further comprises a fifth scanning device 310, the fifth scanning device 310 is used for scanning the identification codes of the deep-hole plates 201-206 filled with the extracted product nucleic acid and the identification codes of the PCR reagent plates 301, and the PCR reaction system constructor acquires and operates a PCR reaction system construction experiment program through the identification codes of the PCR reagent plates 301; the information processing apparatus 6 transfers the sample information recorded in the identification code of the deep-well plate 201 to 206 filled with the extracted product nucleic acid to the identification code of the reaction system PCR plate 302.
The computer 400 is provided with an external sixth scanning device 401 for scanning the identification code of the PCR plate 302 of the reaction system, and the information processing device 6 is used for corresponding the sample information to the analysis result of the detection data.
The sample storage module 5 further comprises a seventh scanning device 501 for scanning the sample rack 102 identification code, the sample cabinet number and the position code of the rack position; the information processing device 6 is used for corresponding the sample rack 102 identification code, the sample cabinet number and the position code of the rack position; the analysis result of the nucleic acid detection data is fed back to the sample cabinet management unit through the information processing device 6.
In this embodiment, the PCR reagent sub-packaging apparatus further includes: a reagent and premixing zone, a Tip zone, a waste zone and a PCR reagent loading zone; the reagent and the premixing zone are used for placing reagents required by the PCR system and premixing the reagents; the Tip area is used for loading samplers with different specifications; the waste area is used for accommodating an automatically unloaded sampler; the PCR reagent loading area is used for placing a PCR plate containing a PCR reagent system.
In this embodiment, the PCR reaction system builder further includes: the system establishment area, the Tip area, the product extraction area, the waste area and the product sample reserving area; the system establishment area is used for placing a PCR reagent plate filled with a PCR reagent, and the Tip area is used for loading samplers with different specifications; the extraction product area is used for loading a deep hole plate filled with extraction products; the waste area is used for accommodating an automatically unloaded sampler; the product sample retaining area is used for retaining the residual extracted product, and is convenient for rechecking.
In the embodiment, the model of the deep hole plate is a 96-hole deep hole plate with the size of 8X 12, arabic numerals 1-12 are arranged on the upper surface of the deep hole plate along the X axis according to the hole position, and English letters A-H are arranged along the Y axis according to the hole position; the hole site distribution format on the sample holder 102 is 8 x 12; the PCR reagent plate and the reaction system PCR plate are 96 well plates with hole site distribution formats of 8X 12, wherein Arabic numerals 1-12 are arranged on the upper surfaces of the 96 well plates along the X axis according to the hole sites, and English letters A-H are arranged along the Y axis according to the hole sites.
In this embodiment, 48 sample rack positions are provided for each of the sample cabinets 50, and 1 to a plurality of sample cabinets can be used in series. The sample cabinet management unit 502 includes the sample rack position display, the positive sample detection list and the abnormal detection result list in the sample cabinet, and displays the marks of the sample racks in the sample cabinet. Specifically, the position of the sample cabinet, where the sample rack is not placed, is marked by white; the sample has been placed and the locations on the sample holder where the sample is being tested and not the result are marked with blue; the samples placed on the positions of the sample racks are subjected to experiments, and the output results are all normal results, wherein the positions are marked with green, and the experimenters can take the samples off the racks; if the sample with abnormal detection result appears on the sample rack, the sample rack at the position is marked with orange.
In this embodiment, the source of the nucleic acid sample is one of a washing solution, plasma, serum, and a cell preservation solution.
In this embodiment, the information processing device is a cloud server.
The traceable automatic nucleic acid detection system provided by the application can be used for combining and assembling a plurality of groups of instruments according to different detection requirements, and particularly, the application provides two design schemes, namely a basic scheme and a high-flux scheme.
The basic scheme provided by the embodiment has a theoretical treatment flux of 10000 tubes/24 hours, and the specific instrument is configured as follows: 3 sample preprocessors (model: geneMix Pro), 3 nucleic acid extractors (model: geneRotex 96), 1 PCR reagent split charging instrument (model: PANA S401), 1 PCR reaction system constructor (model: PANA S401), 10 nucleic acid detectors (model: gentier 96).
The high throughput protocol provided in this example, with a theoretical throughput of 80000 tubes/24 hours, was specifically configured to: 12 sample pretreatment instruments (model number: geneMix Pro), 12 nucleic acid extraction instruments (model number: geneRotex 96), 2 PCR reagent split charging instruments (model number: PANA S401), 6 PCR reaction system construction instruments (model number: PANA S401), 60 nucleic acid detection instruments (model number: gentier 96). In addition, the experimenter can set the instrument amount of corresponding multiple on the basis of the actual processing requirement.
The specific working mode is as follows:
the sample size of the single treatment of this example was set to 96 according to the single maximum treatment flux of the instrument.
Referring to fig. 7-8, when performing large-scale nucleic acid detection, an experimenter receives a large number of detection sample tubes, firstly, the experimenter collects sample tube 101 identification code information and sample frame 102 identification code information by using a first scanning device 103 to obtain sample tube identification code information S1, S2, S3 … … S96 and sample frame identification code information, a position sensor is arranged on the sample frame 102, the position sensor endows the sample tubes S1-S96 with position information L1-L96, and the information processing device 6 records the sample tube identification code information S1-S96, the sample frame identification code information and the position information L1-L96 of the sample tubes S1-S96 on the sample frame, and corresponds the sample tube identification code information S1-S96 with the sample frame identification code information, and the sample tube identification code information S1-S96 corresponds with the position information L1-L96; the information processing device 6 transmits the sample automated transfer test program to the sample preprocessor 20, the PCR reagent system preparation test program to the PCR reagent dispensing instrument, and the nucleic acid detection test program to the nucleic acid detector 40, according to the number of the entered sample tube 101 identification codes.
The sample rack 102 carries samples to be tested into the sample pretreatment instrument 20 for automatic opening/closing cover of the samples, sample transfer, deep hole plate information input, sample identification code identification, sample tube recovery to the sample rack, loading of the samples to the deep hole plate and the like. Specifically, the sample pre-processor 20 transfers the S1-S96 samples on the sample rack 102 to the deep hole plates 201-206, the samples S1-S16 on the sample rack are added to the sample adding hole site 13 of the first deep hole plate with the identification code of 201, the samples S17-S32 are added to the sample adding hole site 13 … … of the second deep hole plate with the identification code of 202, and so on, so that the hole site A1 of the sample S1 corresponding to the first deep hole plate 201, the hole site B1 … … of the sample S2 corresponding to the deep hole plate 201, the hole site A7 of the sample S9 corresponding to the first deep hole plate 201, the hole site B7 … … of the sample S10 corresponding to the first deep hole plate 201, the hole site A1 of the sample S17 corresponding to the second deep hole plate 202, and the hole site B1 … … of the sample S18 corresponding to the second deep hole plate 202 can be obtained; meanwhile, the second scanning device 2011 records the identification code information of the sample rack 102, the identification codes of the S1-S96 sample tubes and the identification codes of the deep hole plates 201-206, the information processing device 6 records the S1-S16 sample information into the identification code of the first deep hole plate 201, the S17-S32 sample information into the identification code of the second deep hole plate 202, and the S81-S96 sample information into the identification code of the sixth deep hole plate 206; meanwhile, the information processing apparatus 6 transmits the nucleic acid extraction experimental procedure to the identification code of each of the deep well plates 201 to 206.
After the sample pretreatment apparatus 20 runs the sample automatic transfer experiment program, an experimenter takes out the sample rack 102 carrying the sample tubes and puts the sample rack into the sample cabinet 50 for storage and arrangement; specifically, the experimenter records the sample cabinet code, the sample rack identification code, and the position code of the sample rack in the sample cabinet by using the seventh scanning device 501 of the sample cabinet 50, and the information processing device 6 corresponds the sample cabinet code, the sample rack identification code, and the position code of the sample rack in the sample cabinet.
Subsequently, the deep-hole plates 201 to 206 are transferred to the nucleic acid extractor 21, the third scanning device 2111 scans the identification code of each deep-hole plate 201 to 206 to obtain sample information, at the same time, the nucleic acid extractor 21 obtains and runs a nucleic acid extraction experimental program, according to the extraction principle of the nucleic acid extractor, the sample is transferred from the sample addition hole site 13 to the extraction product hole site 14, as shown in fig. 8, specifically, the sample S1 is transferred to the hole site A5 of the first deep-hole plate 201, the sample S2 is transferred to the hole site B5 … … of the first deep-hole plate 201, the sample S9 is transferred to the hole site a11 of the first deep-hole plate 201, the sample S17 is transferred to the hole site B11 … … of the first deep-hole plate 201, the sample S17 is transferred to the hole site A5 of the second deep-hole plate 202, and the sample S18 is transferred to the hole site B5 … … of the second deep-hole plate 202, and finally the deep-hole plates 201 to 206 packed with the extraction product nucleic acid are obtained.
The PCR reagent split charging instrument 30 receives the PCR reagent system preparation experimental program transmitted from the information processing device 6, and realizes PCR reagent premixing, PCR reagent transferring and PCR reagent split charging, and the PCR reagent is split-charged in a 96-hole PCR reagent plate with a mark code of 301; meanwhile, the information processing device 6 transmits the PCR reaction system construction experiment program to the identification code of the PCR reagent plate 301;
the experimenter puts deep hole plates 201-206 with the extracted products and PCR reagent plates 301 together into a PCR reaction system constructor 31, and a fifth scanning device 310 of the PCR reaction system constructor 31 acquires and operates a PCR reaction system construction experiment program by scanning the identification codes of the PCR reagent plates 301, and the specific adding sequence is shown in fig. 8: samples S1-S8 are sequentially added to the hole sites A1-H1 of the reagent plate 301, samples S8-S16 are sequentially added to the hole sites A2-H2 … … of the reagent plate 301, and the rest samples are analogized sequentially; in addition, the information processing device 6 transmits the sample information recorded in the identification codes of the deep well plates 201 to 206 to the identification codes of the PCR plates 302 forming the reaction system; the reaction system PCR plate 302 is identical to the PCR reagent plate 301.
The nucleic acid detector 40 acquires and runs a nucleic acid amplification and detection experiment program through the information processing device 6, completes the nucleic acid detection of a sample to be detected, outputs detection data to the computer end 400, analyzes and sorts the detection data by the computer end 400, and then uploads a data analysis result to the information processing device 6; the experimenter scans the identification code of the reaction system PCR plate 302 by using the sixth scanning device 401, and uploads the identification code to the information processing device 6, and the information processing device 6 correlates the sample information with the analysis result information.
After the information processing device 6 acquires the analysis result, the analysis result information is fed back to the sample cabinet management unit 502, and if an abnormal result sample appears in the samples S1-S96 in the sample rack 102, the sample cabinet management unit 502 marks the sample rack as orange; the experimenter directly takes out the sample frame with the abnormal result, according to the nucleic acid detection analysis result, the corresponding relation between the hole site in the reaction system PCR plate 302 and the sample can be obtained, and the sample corresponding to the abnormal detection result can be obtained. If all the analysis results of the sample detection data in S1-S96 in the sample rack 102 are normal, the sample cabinet management unit 502 marks the sample rack as green; the experimenter can directly put the sample rack off the rack for treatment.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, and alternatives falling within the spirit and principles of the invention.
Claims (18)
1. A traceable automated nucleic acid detection system, the automated nucleic acid detection system comprising:
The sample information acquisition module (1), the sample information acquisition module (1) is used for realizing sample information input and uploading of a sample to be detected;
a nucleic acid extraction module (2), wherein the nucleic acid extraction module (2) is used for realizing nucleic acid extraction of a sample to be detected;
the PCR system preparation module (3) is used for adding the extracted product nucleic acid into a PCR reagent system to complete the construction of a PCR reaction system;
a nucleic acid detection module (4), wherein the nucleic acid detection module (4) is used for amplifying and detecting nucleic acid to be detected;
and the information processing device (6) is used for completing information recording, processing and transmitting of each stage in the sample processing process.
2. The automated nucleic acid detection system of claim 1, wherein the sample information acquisition module (1) comprises a sample tube (101), a sample rack (102), and a first scanning device (103);
the sample tube (101) is placed above the sample frame (102), and identification codes are respectively arranged on the sample tube (101) and the sample frame (102);
a position sensor is further arranged on the sample frame (102), and position information acquisition and uploading of each sample tube (101) on the sample frame (102) are completed through the position sensor; the first scanning device (103) is used for realizing acquisition and uploading of identification codes of the sample tube (101) and the sample rack (102);
And the information processing device (6) is configured to realize the correspondence of the sample tube identification code information and the sample frame identification code information and complete the correspondence of the sample tube identification code information and the position information of the sample tube.
3. The automated nucleic acid detection system of claim 1, wherein the nucleic acid extraction module (2) comprises a sample pre-processor (20) and a nucleic acid extractor (21),
the sample pretreatment instrument (20) is used for completing automatic transfer of a sample to be tested; the nucleic acid extraction instrument (21) is used for realizing nucleic acid extraction of a sample to be detected in the sample pretreatment instrument (20).
4. An automated nucleic acid detection system according to claim 3, wherein the sample pre-processor (20) comprises a plurality of deep well plates with identification codes and a second scanning device (2011), the nucleic acid extractor (21) further comprising a third scanning device (2111);
the deep hole plate is used for sub-packaging the transferred sample to be tested, and the second scanning device (2011) is used for completing collection and uploading of the identification codes of the sample tube (101), the sample rack (102) and the deep hole plate;
the identification code of the deep hole plate records sample information of each sample to be detected which is grasped by the information processing device (6) and an experimental program of the nucleic acid extractor (21);
The nucleic acid extraction instrument (21) performs nucleic acid extraction based on an experimental procedure obtained by scanning the identification code of the deep well plate by the third scanning device (2111).
5. The automated nucleic acid detection system of claim 1, wherein the PCR system formulation module (3) comprises a PCR reagent dispensing instrument (30) and a PCR reaction system constructing instrument (31),
the PCR reagent split charging instrument (30) acquires a PCR reagent system preparation program through the information processing device (6) to complete the preparation of the PCR reagent system; the PCR reaction system construction instrument (31) is used for adding the extracted product nucleic acid into a PCR reagent system to complete the construction of the PCR reaction system.
6. The automated nucleic acid detection system of claim 5, wherein the PCR reagent dispensing instrument (30) comprises a fourth scanning device (300) and a plurality of PCR reagent plates (301);
the PCR reagent plate (301) is used for realizing premixing and split charging of the PCR reagents, and the identification code of the PCR reagent plate (301) records the experimental program of the PCR reaction system constructor (31); the fourth scanning device (300) is used for scanning the identification code of the PCR reagent plate (301);
the PCR reaction system construction instrument (31) further comprises a fifth scanning device (310), wherein the fifth scanning device (310) is used for scanning the identification codes of the deep hole plates filled with nucleic acid and the identification codes of the PCR reagent plates (301), and the PCR reagent plates (301) containing samples for completing the construction of the PCR reaction system are used as reaction system PCR plates (302);
The information processing device (6) transmits the sample information recorded in the identification code of the deep-hole plate filled with the nucleic acid to the identification code of the reaction system PCR plate (302).
7. The automated nucleic acid detection system of claim 6, wherein the PCR reagent dispensing instrument (30) further comprises: a reagent and premixing zone, a Tip zone, a waste zone and a PCR reagent loading zone;
the reagent and the premixing area are used for placing reagents required by the PCR reagent system and premixing the reagents; the Tip region is used for loading samplers with different specifications; the waste area is used for accommodating an automatically unloaded sampler; the PCR reagent loading area is used for placing a PCR reagent plate (301) containing a PCR reagent system.
8. The automated nucleic acid detection system of claim 6, wherein the PCR reaction system builder (31) further comprises: the system establishment area, the Tip area, the product extraction area, the waste area and the product sample reserving area;
the system establishment area is used for placing a PCR reagent plate (301) filled with a PCR reagent, and the Tip area is used for loading samplers with different specifications; the extraction product region is used for loading a deep hole plate filled with extraction product nucleic acid; the waste area is used for accommodating an automatically unloaded sampler; the product retention zone is used to retain the remaining extracted product.
9. The automated nucleic acid detection system of claim 6, wherein the nucleic acid detection module (4) comprises a nucleic acid detector (40), a computer terminal (400), and a sixth scanning device (401),
the computer end (400) is used for analyzing and arranging detection data of the nucleic acid detector (40) and uploading analysis results to the information processing device (6);
the sixth scanning device (401) is used for scanning the identification code of the reaction system PCR plate (302), and the information processing device (6) is used for corresponding sample information to the analysis result of the detection data.
10. The automated nucleic acid detection system of claim 1, further comprising a sample receiving module (5), the sample receiving module (5) being configured to receive and manage sample tubes for which sample transfer has been completed.
11. The automated nucleic acid detection system of claim 10, wherein the sample receiving module (5) comprises a plurality of sample cabinets (50), each sample cabinet (50) having a position-coded rack location for receiving a sample rack (102);
each sample cabinet (50) further comprises a seventh scanning device (501) and a sample cabinet management unit (502), the sample cabinet management unit (502) is used for completing the visual display of the sample detection state on the sample rack (102) in the sample cabinet (50) and the position of the sample rack (102),
The seventh scanning device (501) is used for scanning the identification code of the sample rack (102), the number of the sample cabinet (50) and the position code of the rack position; the identification code of the sample rack (102), the number of the sample cabinet (50) and the position code of the rack position are respectively corresponding through the information processing device (6); the analysis result of the nucleic acid detection data is fed back to the sample cabinet management unit (502) through the information processing device (6).
12. A traceable automated nucleic acid detection method, the automated nucleic acid detection method comprising:
step 1: a sample information input step, wherein the sample information comprises identification code information of a sample tube (101), a sample frame (102) and position information of the sample tube (101) on the sample frame (102);
step 2: a pre-treatment step of the sample to be measured, completing automatic transfer of the sample to be measured;
step 3: extracting nucleic acid of a sample to be detected;
step 4: preparing a PCR system; adding the extracted product nucleic acid into a PCR reagent system to complete the construction of a PCR reaction system;
step 5: a nucleic acid detection step of amplifying and detecting the nucleic acid to be detected;
step 6: and a sample tube storage management step of storing and managing the sample tubes with the samples transferred, and displaying the analysis results of the nucleic acid detection data of each sample tube.
13. The automated nucleic acid detection method of claim 12, wherein the sample information entry step of step 1 comprises:
the first scanning device (103) scans the identification code information on the sample tube (101) and the sample frame (102) and uploads the identification code information to the information processing device (6), meanwhile, the position sensor on the sample frame (102) uploads the recorded position information of the sample tube (101) to the information processing device (6), and the information processing device (6) corresponds the identification code information on the sample tube (101), the identification code information on the sample frame (102) and the position information of the sample tube (101) on the sample frame (102); the information processing device (6) respectively sends a sample automatic transfer experiment program, a PCR reagent system preparation program and a nucleic acid detection experiment program to the sample pretreatment instrument (20), the PCR reagent split charging instrument (30) and the nucleic acid detection instrument (40) according to the number of the recorded identification codes of the sample tubes (101).
14. The automated nucleic acid testing method of claim 13, wherein the pre-processing step of the test sample of step 2 comprises:
automatically transferring the sample to be tested in the sample tube (101) to a deep hole plate with an identification code by adopting a sample pretreatment instrument (20) based on a sample automatic transfer experimental program;
The second scanning device (2011) scans the identification code information of the sample tube (101), the identification code information of the sample rack (102) and the identification code of the deep hole plate, and the information processing device (6) transmits the sample information to the identification code of the deep hole plate to realize the correspondence between the identification code of the deep hole plate and the sample information; meanwhile, the nucleic acid extraction experimental program is transferred to the identification code of the deep well plate through the information processing device (6).
15. The automated nucleic acid detection method of claim 14, wherein the nucleic acid extraction step of the test sample of step 3 comprises:
the third scanning device (2111) scans the identification code of the deep hole plate, the nucleic acid extractor (21) acquires and runs a nucleic acid extraction experiment program through the identification code of the deep hole plate, nucleic acid extraction is carried out on a sample to be detected transferred into the deep hole plate, and sample information recorded in the identification code of the deep hole plate is grasped.
16. The automated nucleic acid detection method of claim 13, wherein the PCR system formulation step of step 4 comprises:
the PCR reagent split charging instrument (30) acquires a PCR reagent system preparation program through the information processing device (6) to complete the preparation of the PCR reagent system; meanwhile, the fourth scanning device (300) scans the identification code of the PCR reagent plate (301) filled with the PCR reagent system and uploads the identification code to the information processing device (6), and the information processing device (6) records the PCR reaction system construction experiment program into the identification code of the PCR reagent plate (301);
Then, a fifth scanning device (310) scans the identification code of the PCR reagent plate (301) and the identification code of the deep hole plate containing the extracted product nucleic acid, and uploads the identification code to an information processing device (6), a PCR reaction system constructing instrument (31) acquires and operates a PCR reaction system constructing experiment program to complete the preparation of a PCR reaction system, and a sample constructed by the PCR reaction system is contained in a reaction system PCR plate (302); the information processing device (6) transmits sample information corresponding to the identification code of the deep well plate containing the nucleic acid to the identification code of the reaction system PCR plate (302).
17. The automated nucleic acid detection method of claim 13, wherein the nucleic acid detection step of step 5 comprises:
the nucleic acid detector (40) acquires and runs a nucleic acid detection experiment program through the information processing device (6), completes nucleic acid detection of a sample to be detected, outputs detection data to the computer end (400), analyzes and sorts the detection data by the computer end (400), and then uploads a data analysis result to the information processing device (6); the sixth scanner (401) scans the identification code of the reaction system PCR plate (302) and uploads the identification code to the information processing device (6), and the information processing device (6) correlates the data analysis result with the sample information.
18. The automated nucleic acid testing method of claim 12, wherein the sample tube storage management step of step 6 comprises:
a seventh scanning device (501) scans the identification code information of the sample rack (102) carrying the sample tubes (101), the number of the sample cabinet (50) and the position code of the sample rack position, and places the sample rack (102) carrying the sample tubes (101) on the rack position in the sample cabinet (50); the information processing device (6) feeds the analysis result of the sample detection data back to the sample cabinet management unit (502), and the sample cabinet management unit (502) performs visual display on the position of the sample frame (102) and the analysis result of the detection data of the sample on the sample frame (102).
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