CN117925384A - Blood nucleic acid screening assembly line system - Google Patents
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Abstract
The invention belongs to the technical field of blood screening, and particularly relates to a blood nucleic acid screening pipeline system. The system mainly comprises nine large instrument modules and an integrated laboratory information management system, wherein the instrument modules comprise an ATM self-service handover module, a sample screening module, an online centrifugation module, a sample uncapping module, a sample transferring module, a sample collecting module, a sample extracting module, a sample capping module and a cold storage and preservation module, and the modules are connected through rails and are communicated and independently controlled through matched software. The system can realize the full-flow automation of blood sieve nucleic acid detection.
Description
Technical Field
The invention belongs to the technical field of blood screening, and particularly relates to a blood nucleic acid screening pipeline system.
Background
The prevention and control of infectious diseases related to blood transfusion are the focus of national and global attention, the nucleic acid amplification detection technology is one of the most sensitive and most advanced virus detection technologies so far, can obviously shorten the window period, reduce the transmission risk of blood-borne viruses, and is an important and effective supplement to the traditional immunological detection means. Perfecting the gratuitous blood donation service system, improving the blood safety guarantee capability of the blood station, actively promoting the nucleic acid detection work of the blood station, improving the laboratory detection level of the blood station, and basically realizing the full coverage of the nucleic acid detection by the national blood station. There are 9 current nucleic acid detection systems in nationwide blood stations, of which there are 3, grifols, perkinElmer, roche, 6, kowa, hua Yi m, dar, mo Tai, san xiang, li zhu. The domestic brand is basically developed on the basis of a Hamilton or Tecan pipetting workstation platform to complete the full-automatic nucleic acid extraction and system construction work of samples, and the imported brand part can realize the integration of full-automatic nucleic acid extraction and detection.
At present, the blood nucleic acid screening system of the blood station in China cannot realize the automation of the whole detection process, including import and domestic brands, sample extraction automation or extraction and detection integration are realized, a sample pretreatment part before detection comprises sample handover, sample screening and classification, sample storage before detection, sample centrifugation, blood sampling tube cover opening and sample rack loading, a plurality of links involved need a large number of manual participation to be completed, a sample post-treatment part after detection comprises an unloading sample carrier, sample tube cover loading, sample preservation after detection and the like also need manual participation to be completed, and the steps are complicated, manual operation is more, each link is easy to have errors, the work efficiency is influenced, the cost is increased, the whole process can not be traced, and the timeliness and the accuracy of the detection result are seriously influenced.
Therefore, development of a detection system capable of realizing full-process automation, intellectualization and informatization of blood sieve nucleic acid detection is needed, and an intelligent blood station is actively created for blood safety maintenance and navigation in response to the guidance of national intelligent medical policies.
Disclosure of Invention
The invention aims to provide a production line system capable of realizing full-flow automation of blood nucleic acid screening for a blood station laboratory, which is used for connecting different modules through rails in the processes of sample handover, sample sorting, sample preservation before detection, sample centrifugation, sample uncapping, sample transferring, sample collection, sample extraction, sample capping, sample preservation after detection and the like, organically integrating, realizing full-flow automation of detection, realizing traceability in the whole process and ensuring the accuracy of a nucleic acid detection result.
The invention adopts the following technical means:
a blood nucleic acid screening pipeline system comprising the following modules:
the ATM self-service handover module is used for completing system login, sample type identification, counting the number of handover samples and printing handover sheets in a self-service manner;
the sample screening module is used for identifying and inputting the detailed information, grabbing and moving the samples and screening and sorting the samples;
The sample centrifugation module is used for centrifugal processing and balancing operation;
the sample uncapping module is used for uncapping and storing the sample tube and monitoring the state and the position of the sample tube;
The sample transfer module is used for conveying and returning samples;
the sample mixing module is used for mixing samples and detecting the liquid level of the samples;
The sample extraction module is used for full-automatic extraction of sample nucleic acid and a PCR system component;
The sample capping module is used for capping the uncapped sample with the sucked sample;
the sample refrigerating and preserving module is used for refrigerating and preserving samples and automatically returning the primary screening positive samples to the sample screening module;
A laboratory information management system;
the modules are connected through a track and are communicated and independently controlled through matched software.
Further, the ATM self-service delivery module includes: the device comprises a touch display screen, a keyboard, a fingerprint identification device, an ID card identification device, a printer and a CCD photographing module for self-help login, sample type identification and sample count and handover;
Further, the sample screening module includes: the system comprises a platform base, a plurality of sample supporting stations, an abnormal sample processing station, a movable mechanical arm, a mechanical gripper assembly and a bar code scanning assembly, wherein the platform base can freely move in the X/Y/Z axis direction, samples can be captured and 360-degree omnibearing scanning can be realized;
further, the sample centrifugation module comprises: the device comprises a platform base, a low-temperature centrifuge below the base and a balancing device on the base;
Further, the sample door module includes: the movable mechanical arm and the gripper are configured to realize free movement in the X/Y/Z axis direction, and a waste box device is arranged below the base;
Further, the sample transfer module comprises: the platform base, the movable mechanical arm, the gripper and the sample transmission chains can realize the transportation of the sample after the cover is opened;
Further, the sample mixing module includes: the device comprises a platform base and a movable mechanical arm above the base, wherein a plurality of sample adding channels with a liquid level detection function are arranged on the mechanical arm, and the mechanical arm can realize free movement in the X/Y/Z axis direction;
further, the sample extraction module includes: the platform base is provided with a temperature control oscillation module, a magnetic attraction module, a deep hole plate carrier, a gun head carrier module, a reagent tank module and a waste liquid box;
further, the sample capping module comprises: a consumable storage device and a capping device;
further, the sample cryopreservation module comprises: a front refrigeration module and a rear refrigeration module.
Further, in the sample screening module, the liquid level detection is performed by a capacitive or pneumatic method.
Further, the sample extraction module further comprises an extraction platform, wherein the extraction platform comprises a consumable part area, a reagent area, an extraction area and a waste liquid area, and the consumable part area is used for placing a gun head carrier; the reagent zone is provided with a reagent tank carrier for storing a nucleic acid extraction reagent and a PCR reaction solution; the extraction area is provided with a temperature control vibration mixing module and a magnetic absorption module, and is used for magnetic separation and purification of nucleic acid; the waste liquid area is provided with a waste liquid groove for storing waste liquid generated in the extraction process.
Further, the blood nucleic acid screening pipeline system comprises the following workflow:
(1) The ATM automatically hands over the sample, pre-processes and inputs bar code information, and centrifugally and pre-refrigerates and stores the sample to reserve an experiment; the centrifugation step is optional;
(2) Carrying out uncovering operation from front refrigerating when the reserved time sample is reached;
(3) The uncapped sample is transmitted to a sample mixing and extracting module for sample suction and mixing, nucleic acid extraction and PCR system construction;
(4) After the sample is sucked, the sample is conveyed to a capping module;
(5) The capping module performs capping operation;
(6) Transferring the capped sample to a post-refrigeration storage;
(7) PCR amplification, data analysis and report transmission
Screening from a rear refrigeration module for the presence of pooled positive wells or samples to be retested;
(9) The positive sample to be split can enter the next round of experiment retest.
Furthermore, if the detection result prompts that retest or splitting is needed, the retest or splitting specimen can be automatically selected through the experimental script.
The invention has the following beneficial effects:
(1) According to the invention, a plurality of instrument function modules are developed according to pain points and demands of the existing clients, and the modules are organically integrated through control software innovation, so that the automation of the whole blood sieve nucleic acid detection process is realized, a traditional series of complicated manual operation processes are integrated into one production line, the simplicity is simplified, the human error is reduced, and the more standardized operation process is realized.
(2) And the Laboratory Information Management System (LIMS) is matched for integrated development, so that informatization of laboratory management is realized, the whole process is traceable, and the accuracy of a nucleic acid detection result is ensured.
(3) At present, about 80% of manpower consumption in a laboratory is used in the sample detection pretreatment and detection post-treatment processes, and the scheme adopts a pipelining mode, so that the manpower is liberated, the labor intensity is reduced, the experimental efficiency is improved, the occupational exposure is reduced, and the biological risk is reduced.
(4) The pipeline supports a custom reservation detection function, can automatically run at night or in non-working time, is more fully utilized in time, and reduces running cost and time cost.
Drawings
FIG. 1 is a schematic diagram of an embodiment of the present invention;
fig. 2 is a flow chart illustrating the operation of an embodiment of the present invention.
Wherein each figure is numbered:
1. An ATM self-service handover module; 2. a sample centrifugation module; 3. a sample screening module; 4. a sample uncapping module; 5. a sample capping module; 6. a cold storage module; 7. a sample transfer module; 8. a sample mixing module; 9. and a sample extraction module.
Detailed Description
The following description of the embodiments of the present invention will be made with reference to the accompanying drawings in a clear and complete manner with reference to the technical solutions in the examples.
As shown in fig. 1, the blood nucleic acid screening pipeline system is composed of a plurality of main modules, namely 1-9, and comprises an ATM self-service handover module 1, a sample screening module 3, a sample centrifugation module 2, a sample uncapping module 4, a sample transferring module 7, a sample collecting/mixing module 8, a sample extracting module 9, a sample capping module 5 and a cold storage module 6, wherein the modules are connected through rails. Integrated Laboratory Information Management System (LIMS).
In the embodiment, the ATM self-service handover module 1 is provided with a printer in a module box, the right side of the box is provided with a CCD photographing module, system login, sample type identification, sample count and handover quantity and handover list printing can be completed at the interface in a self-service manner, the sample holders take standard centrifugal holders as carriers, 20 sample tubes can be loaded in each sample holder, and the handover speed per hour is more than or equal to 1000 sample tubes/h.
In this embodiment, if the sample is required to be centrifuged twice, the sample screening module 3 performs on-line centrifugation according to the centrifugation module configuration 2. The sample screening platform is provided with more than 30 stations and can support simultaneous screening of 600 sample tubes and 4 centrifugally balancing sample stations. In the workflow, a plurality of choices are provided for the customer, for example, if no experiment is performed on the same day, the customer can choose to directly enter the cold storage and preservation module 6 for temporary storage, and the time for starting detection is set for reservation; if the detection is needed immediately, the sample can enter a screening module for sample bar code identification and input, and the sorting speed of enzyme-free positive samples is more than or equal to 1000 counts/h.
In this embodiment, sample uncapping module 4, sample letter sorting is over, confirms that the sample that needs to carry out the nucleic acid screening gets into uncapping platform, installs carousel formula uncapping machine on the platform, and the waste box is installed to the platform lower part for the storage sample lid, simultaneously material loading manipulator and unloading manipulator, two manipulators are accomplished the uncapping transfer process of sample pipe in coordination.
In this embodiment, the sample is transported the module 7, and the sample of uncapping completion is transferred to the transport module by unloading manipulator, and transport the module and adopt two-way chain transmission, can realize the transportation and the return of sample, accomplish the imbibition when the sample, then return from the opposite side to by unloading manipulator transfer to the sample hold in the palm in. The transportation and the uncapping are synchronously completed, so that 8-sample uncapping and transmission are adopted to avoid cross contamination, and the risk of cross contamination caused by exposing a large number of samples to the uncapping at the same time is avoided to the greatest extent.
In this embodiment, the sample uncapped by the sample mixing module 8 is sent to the designated liquid absorbing position through the transfer and conveying module, the sample adding gun of the 8 channels configured by the sample mixing module performs liquid level detection in a capacitive (cLLD) or pneumatic (pLLD) mode and has the abnormal sample processing functions of air absorption, clot detection and the like, so that accurate transfer of the sample is completed, the sample is transferred to 2.0ml 96-hole deep hole plates of two standards, and according to a 6-collection mode, collection of 576 samples can be completed at maximum, and a total of 96 collection holes are formed. And for the single-sample, completing the transfer of the single-sample of 1 to 1 according to a1 aggregation mode.
In this embodiment, the sample extraction module 9, after completing the transfer of the collected and single samples, enters a fully automatic extraction platform, and the extraction platform has a consumable part area, a reagent area, an extraction area and a waste liquid area. The consumable area can be used for placing a gun head carrier, and the carrier can be used for loading 1000ul and 200ul of disposable Tips; the reagent area is provided with a reagent groove carrier for storing nucleic acid extraction reagent and PCR reaction liquid components, including a lysate, a washing liquid A, a washing liquid B, an eluent, a magnetic bead suspension, a digestion liquid, an internal standard, polyA, a PCR reaction liquid A/a PCR reaction liquid B/a PCR reaction liquid C and the like; the extraction area is provided with a temperature control vibration mixing module and a magnetic absorption module, the magnetic separation and purification of nucleic acid are mainly completed, and the waste liquid area is provided with a waste liquid tank for storing waste liquid generated in the extraction process. Through reasonable setting of the partition, reasonable planning of the pipetting path satisfies the extraction workflow and reduces pollution risk. The extraction and the construction of the PCR system are completed, the next detection flow can be carried out, the LIMS monitors the amplification result, and the integration and the analysis of the result are automatically carried out.
In this embodiment, the sample capping module 5, uncaps and completes the sample of the suction sample, get into the capping module, the capping module is furnished with consumable box and capping device, there is disposable silica gel pearl in the consumable box memory, one capping action, can once only accomplish the capping of 20 sample tubes. The capping speed is more than or equal to 1000 counts/h.
In this embodiment, the sample before and after detection needs to be refrigerated and stored in the refrigeration storage module, and the temperature of the refrigeration storage module is set to 2-8 ℃ without temperature step storage. The detected sample is stored in the post-refrigeration module, after the detection result is obtained, the post-treatment is carried out, and if the detection result prompts that retest or splitting is needed, the retest or splitting sample can be automatically selected through an experimental script.
Example 1
The workflow of the blood nucleic acid screening pipeline system comprises the following steps:
(1) The ATM automatically hands over the sample, pre-processes and inputs bar code information, and centrifugally (optionally) stores the sample in a front refrigerating way, and reserves the experiment;
(2) Carrying out uncovering operation from front refrigerating when the reserved time sample is reached;
(3) The uncapped sample is transmitted to a sample mixing and extracting module for sample suction and mixing, nucleic acid extraction and PCR system construction;
(4) After the sample is sucked, the sample is conveyed to a capping module;
(5) The capping module performs capping operation;
(6) Transferring the capped sample to a post-refrigeration storage;
(7) PCR amplification, data analysis and report transmission
(8) Screening from a rear refrigeration module for the presence of pooled positive wells or samples to be retested;
(9) The positive sample to be split can enter the next round of experiment retest.
Example 2
Application example: the system performance of the blood sieve assembly line system is comprehensively verified by using an international standard (primary reference) and yin-yang samples.
HBV (NIBSC CODE 10/266), HCV (NIBSC CODE 18/184), HIV (NIBSC CODE 16/194), lyophilized powder, and reconstituted with purified water, diluted to a system detection limit concentration, detection system sensitivity, HBV 2.5IU/mL, HCV 9IU/mL, and HIV 20IU/mL. The detection rates are all 100 percent, and all meet the requirements.
HBV (NIBSC CODE 10/266), HCV (NIBSC CODE 18/184), HIV (NIBSC CODE 16/194) and freeze-dried powder are diluted to 10 times detection limit after being re-dissolved by purified water, and the precision of the test system, HBV 25IU/mL, HCV 90IU/mL, HIV 200IU/mL and precision CV less than 5 percent meet the requirements.
The specific operation flow of the verification is as follows:
S1, diluting WHO to detection limit and precision, sub-packaging a detection limit sample into 20 parts, sub-packaging the precision into 10 parts, and detecting according to a normal sample.
S2, placing the sample to be detected on a standard sample carrier, carrying out handover in an ATM handover window, and automatically returning the handover number.
And S3, automatically completing bar code scanning by the sample screening module, and entering the cover opening module after the sample scanning is completed.
S4, the uncapped sample is transmitted to a sample mixing and extracting module through a conveying module to suck the sample, and meanwhile, the extracting module carries out automatic extraction and PCR system construction flow
S5, conveying the sample after the sample is sucked to a capping module
And S6, after capping is completed, the sample enters into the refrigerator for preservation.
And S7, after the extraction and system construction are finished, placing the amplification plate into an amplification instrument for amplification detection, and counting the detection rate.
And (3) verifying pollution resistance, namely adopting a chessboard experiment to prepare high-concentration samples and negative samples which are arranged in a crossing way, counting the coincidence rate of negative and positive results, and normally detecting the positive samples, wherein the negative samples are not jumped, and the result meets the requirement.
The embodiment shows that the blood sieve nucleic acid detection assembly line system has excellent performance, realizes the automation, the intellectualization and the informatization of the whole flow before, during and after the detection, greatly saves the labor cost, improves the working efficiency and is at the leading level at present in China.
It is apparent that the above-described embodiments are only some embodiments of the present invention, but not all embodiments, and the preferred embodiments of the present invention are shown in the drawings, which do not limit the scope of the patent claims. This invention may be embodied in many different forms, but rather, embodiments are provided in order to provide a thorough and complete understanding of the present disclosure. Although the invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described in the foregoing description, or equivalents may be substituted for elements thereof. All equivalent structures made by the content of the specification and the drawings of the invention are directly or indirectly applied to other related technical fields, and are also within the scope of the invention.
Claims (6)
1. A blood nucleic acid screening pipeline system, comprising the following modules: the ATM self-service handover module is used for completing system login, sample type identification, counting the number of handover samples and printing handover sheets in a self-service manner;
the sample screening module is used for identifying and inputting the detailed information, grabbing and moving the samples and screening and sorting the samples;
The sample centrifugation module is used for centrifugal processing and balancing operation;
the sample uncapping module is used for uncapping and storing the sample tube and monitoring the state and the position of the sample tube;
The sample transfer module is used for conveying and returning samples;
the sample mixing module is used for mixing samples and detecting the liquid level of the samples;
The sample extraction module is used for full-automatic extraction of sample nucleic acid and a PCR system component;
The sample capping module is used for capping the uncapped sample with the sucked sample; the sample refrigerating and preserving module is used for refrigerating and preserving samples and automatically returning the primary screening positive samples to the sample screening module;
A laboratory information management system;
the modules are connected through a track and are communicated and independently controlled through matched software.
2. The blood nucleic acid screening pipeline system of claim 1, wherein,
The ATM self-service handover module comprises: the device comprises a touch display screen, a keyboard, a fingerprint identification device, an ID card identification device, a printer and a CCD photographing module for self-help login, sample type identification and sample count and handover;
the sample screening module includes: the system comprises a platform base, a plurality of sample supporting stations, an abnormal sample processing station, a movable mechanical arm, a mechanical gripper assembly and a bar code scanning assembly, wherein the platform base can freely move in the X/Y/Z axis direction, samples can be captured and 360-degree omnibearing scanning can be realized; the sample centrifugation module includes: the device comprises a platform base, a low-temperature centrifuge below the base and a balancing device on the base;
the sample uncap module includes: the movable mechanical arm and the gripper are configured to realize free movement in the X/Y/Z axis direction, and a waste box device is arranged below the base;
the sample transfer module includes: the platform base, the movable mechanical arm, the gripper and the sample transmission chains can realize the transportation of the sample after the cover is opened;
The sample mixing module comprises: the device comprises a platform base and a movable mechanical arm above the base, wherein a plurality of sample adding channels with a liquid level detection function are arranged on the mechanical arm, and the mechanical arm can realize free movement in the X/Y/Z axis direction;
the sample extraction module includes: the platform base is provided with a temperature control oscillation module, a magnetic attraction module, a deep hole plate carrier, a gun head carrier module, a reagent tank module and a waste liquid box;
The sample capping module includes: a consumable storage device and a capping device;
The sample cryopreservation module comprises: a front refrigeration module and a rear refrigeration module.
3. The blood nucleic acid screening pipeline system of claim 1, wherein in the sample screening module, liquid level detection is performed by capacitive or pneumatic means.
4. The blood nucleic acid screening pipeline system of claim 1, wherein the sample extraction module further comprises an extraction platform comprising a consumable region, a reagent region, an extraction region, a waste region, the consumable region housing a gun head carrier; the reagent zone is provided with a reagent tank carrier for storing a nucleic acid extraction reagent and a PCR reaction solution; the extraction area is provided with a temperature control vibration mixing module and a magnetic absorption module, and is used for magnetic separation and purification of nucleic acid; the waste liquid area is provided with a waste liquid groove for storing waste liquid generated in the extraction process.
5. The blood nucleic acid screening pipeline system of claim 1, comprising the workflow of:
(1) The ATM automatically hands over the sample, pre-processes and inputs bar code information, and centrifugally and pre-refrigerates and stores the sample to reserve an experiment; the centrifugation step is optional;
(2) Carrying out uncovering operation from front refrigerating when the reserved time sample is reached;
(3) The uncapped sample is transmitted to a sample mixing and extracting module for sample suction and mixing, nucleic acid extraction and PCR system construction;
(4) After the sample is sucked, the sample is conveyed to a capping module;
(5) The capping module performs capping operation;
(6) Transferring the capped sample to a post-refrigeration storage;
(7) PCR amplification, data analysis and report transmission
(9) Screening from a rear refrigeration module for the presence of pooled positive wells or samples to be retested;
(9) The positive sample to be split can enter the next round of experiment retest.
6. The blood nucleic acid screening pipeline system of claim 5, wherein if the detection result indicates that retesting or splitting is required, retesting or splitting specimens can be automatically selected by the experimental script.
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| CN202311775958.3A CN117925384A (en) | 2023-12-21 | 2023-12-21 | Blood nucleic acid screening assembly line system |
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| CN202311775958.3A CN117925384A (en) | 2023-12-21 | 2023-12-21 | Blood nucleic acid screening assembly line system |
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