WO2021219067A1 - 流水线核酸处理设备和流水线分子诊断设备 - Google Patents

流水线核酸处理设备和流水线分子诊断设备 Download PDF

Info

Publication number
WO2021219067A1
WO2021219067A1 PCT/CN2021/090890 CN2021090890W WO2021219067A1 WO 2021219067 A1 WO2021219067 A1 WO 2021219067A1 CN 2021090890 W CN2021090890 W CN 2021090890W WO 2021219067 A1 WO2021219067 A1 WO 2021219067A1
Authority
WO
WIPO (PCT)
Prior art keywords
module
pipeline
temperature
nucleic acid
samples
Prior art date
Application number
PCT/CN2021/090890
Other languages
English (en)
French (fr)
Inventor
叶宝春
吴元吉
王珺
黄飞
Original Assignee
杭州杰毅生物技术有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 杭州杰毅生物技术有限公司 filed Critical 杭州杰毅生物技术有限公司
Publication of WO2021219067A1 publication Critical patent/WO2021219067A1/zh

Links

Images

Classifications

    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids

Definitions

  • the present invention relates to the field of nucleic acid detection, in particular to pipeline nucleic acid processing equipment and pipeline molecular diagnostic equipment.
  • nucleic acid extraction and purification Taking the fluorescent quantitative PCR routinely carried out in hospitals as an example, the operation steps include nucleic acid extraction, reaction configuration, sealing film, setting up PCR programs, and amplification.
  • the current design is that multiple samples are operated at the same time, which is convenient to process a large number of samples at the same time and do the same type of detection, which greatly improves the throughput of molecular detection, and can process 96 or even 384 samples in a few hours. Detection. For large-scale screening test items for HBV, HPV, and HIV, or third-party clinical testing central laboratories, this batch processing method can solve the needs of customers who need to process a large number of samples at the same time.
  • the inventor of the present application developed a molecular diagnostic device based on the pipeline idea.
  • the batch processing method can be replaced by a pipeline method to achieve on-demand inspection.
  • the instrument can automatically determine the running item by scanning the sample to detect the barcode.
  • a pipeline nucleic acid processing equipment including:
  • a driving conveying device for receiving and conveying samples from the entrance of the pipeline nucleic acid processing equipment
  • the control device is used to control the operation of each part of the pipeline nucleic acid processing equipment
  • the mixing module, the magnetic sorting module and the temperature-controlled elution module are arranged along the moving track of the traveling conveyor, and the samples are received and removed by the traveling conveyor.
  • the mixing module, the magnetic separation module and the temperature-controlled elution module are arranged in sequence along the moving track of the traveling conveyor.
  • the mixing module is arranged at an upstream position of the moving track of the traveling conveyor.
  • the driving conveying device includes driving positions corresponding to the positions of the mixing module, the magnetic separation module, and the temperature-controlled elution module, and can respectively send the mixing module to the mixing module at the driving position.
  • the magnetic sorting module and the temperature-controlled elution module transmit samples and receive samples from the mixing module, the magnetic sorting module and the temperature-controlled elution module.
  • it further includes a pipetting device arranged at the entrance of the pipeline nucleic acid processing equipment.
  • the driving transport device can receive a sample from the pipetting device.
  • the mixing module, the magnetic separation module and the temperature-controlled elution module can respectively process 1 to 4 samples at the same time.
  • a tip placement device for placing a tip is provided next to each test tube loaded with a sample, and the tip is placed in the tip placement device of the corresponding test tube.
  • a pipelined molecular diagnostic equipment including:
  • a driving conveying device for receiving and transmitting samples from the entrance of the pipeline molecular diagnostic equipment
  • test cartridge with pre-filled reagents for independent temperature control reaction on received samples
  • the optical inspection module detects the delivered inspection cassette
  • the control device is used to control the operation of each part of the pipeline molecular diagnostic equipment
  • the mixing module, the magnetic sorting module and the temperature-controlled elution module are arranged along the moving track of the traveling conveyor, and the samples are received and removed by the traveling conveyor.
  • the mixing module, the magnetic separation module and the temperature-controlled elution module are arranged in sequence along the moving track of the traveling conveyor.
  • the mixing module is arranged at an upstream position of the moving track of the traveling conveyor.
  • the driving conveying device includes driving positions corresponding to the positions of the mixing module, the magnetic separation module, and the temperature-controlled elution module, and can respectively send the mixing module to the mixing module at the driving position.
  • the magnetic sorting module and the temperature-controlled elution module transmit samples and receive samples from the mixing module, the magnetic sorting module and the temperature-controlled elution module.
  • it also includes a pipetting device arranged at the entrance of the pipeline molecular diagnostic equipment.
  • the driving transport device can receive a sample from the pipetting device.
  • the mixing module, the magnetic separation module and the temperature-controlled elution module can respectively process 1 to 4 samples at the same time.
  • a tip placement device for placing a tip is provided next to each test tube loaded with a sample, and the tip is placed in the tip placement device of the corresponding test tube.
  • the detection cartridge is a disposable detection cartridge with a porous tube, has a lid that can be opened and closed, and can control the temperature of the temperature control reaction of the sample received in the disposable detection cartridge and time.
  • the detection content of each sample can be flexibly combined, taking into account higher throughput, flexibility and automation.
  • Fig. 1 is a schematic diagram of a pipelined molecular diagnostic device according to an embodiment of the present invention
  • FIG. 2 is a schematic diagram of the structure of the pipeline molecular diagnostic equipment of FIG. 1;
  • Figure 3 is a schematic diagram of a pipeline nucleic acid processing device according to an embodiment of the present invention.
  • Figure 4 is a schematic diagram of a test tube and tip placement device according to the present invention.
  • Figure 5 is a schematic diagram of a disposable cartridge according to the present invention.
  • 6A and 6B are schematic diagrams of the operation of the driving conveying device according to the present invention.
  • Molecular diagnosis basically includes several links such as nucleic acid extraction, reaction configuration, and detection.
  • the whole process is relatively cumbersome, and it is basically manual use.
  • the nucleic acid extraction is 40 minutes
  • the reaction system is configured for 30 minutes
  • the fluorescent quantitative PCR is two hours.
  • the overall test process is often 4-5 Hours.
  • pipetting workstations represented by companies such as TECAN and Hamilton are the main ones.
  • the general structure principle is basically to add 4, 8 or 96 channel pipette pumps on a water platform, and place liquid and sample storage at the bottom. Supplies. The sample and various reagents are mixed and reacted in the consumable through the pipette pump.
  • the basic consumables are 24, 48, 96 and 384 well plates.
  • the waste liquid and the waste tip are moved to the waste position by the mechanical arm that drives the pipette pump to be discarded.
  • the advantage is that a large number of samples (24, 48, 96, etc.) can be processed at one time to perform the same type of testing, but it is difficult to meet the needs of sample testing at different times.
  • the pipette pump needs to replace the pipette tip after each pipetting and mixing.
  • a simple extraction process involves adding samples, adding lysis solution, sucking waste solution, adding cleaning solution, sucking waste solution, adding cleaning solution, sucking waste solution, adding cleaning solution, sucking waste solution, adding cleaning solution, sucking waste solution, adding eluent and eluting.
  • the washing liquid pump needs to go back and forth between the sample consumables and the pipette tip 10 times, and 10 pipette tips are required.
  • Each replacement is calculated in 1 second, and the lotion pump needs to move 10 back and forth in the extraction process, a total of 10 minutes, accounting for 40% of the entire nucleic acid extraction process.
  • the present invention changes the molecular diagnosis method of conventional batch processing into a pipeline form.
  • the nucleic acid extraction process can be split into three modules: a mixing module, a magnetic sorting module, and a temperature-controlled elution module. Between the three modules, the sample is moved by driving instead of a pipette pump. Although this reduces the throughput, but by breaking up the overall time of nucleic acid extraction, the time for each individual step is about a few minutes. In this way, new samples can be added at any time, shortening the delivery time of clinical results for each individual sample.
  • the extracted sample is pipetted into an independent cassette pre-filled with detection probes.
  • the cassette can be multi-tube, polygonal and other structures, with an automatic lid opening function on the top, and an independent temperature control reaction is completed in the cassette.
  • the optical detection module detects the signal independently.
  • Fig. 1 is a schematic diagram of a pipelined molecular diagnostic device according to an embodiment of the present invention
  • Fig. 2 is a schematic structural diagram of the pipelined molecular diagnostic device of Fig. 1
  • Fig. 3 is a schematic diagram of a pipelined nucleic acid processing device according to an embodiment of the present invention.
  • a pipeline nucleic acid processing device including: a mixing module, a magnetic sorting module, and a temperature-controlled elution module that are separately arranged; A driving conveying device for receiving and conveying samples; and a control device for controlling the operation of each part of the pipeline nucleic acid processing equipment; wherein the mixing module, the magnetic sorting module and the temperature-controlled elution module are conveyed along the driving The moving track of the device is set up, and the sample is received and removed through the driving conveyor.
  • the traveling conveyor device may include traveling vehicles for conveying and a moving track on which the traveling vehicles move.
  • the carriage can be equipped with hooks to grab and unload the reaction cup or sample tube, or other sample loading containers.
  • a screw motor can be used to drag the reaction cup into the driving car through the hook, the driving car rotates to a moving position that is convenient for moving on the moving track, and the pulley translation function is used to move the driving car to the next station or driving position.
  • the claw and screw motor can complete the unloading of the reaction cup.
  • the moving track of the traveling conveyor can be basically linear, or it can also be a track of other linear shapes.
  • the mixing module, the magnetic separation module and the temperature control elution module are arranged in sequence along the moving track of the traveling conveyor.
  • the mixing module is arranged at an upstream position of the moving track of the traveling conveyor.
  • the upstream location usually refers to the location closer to the entrance of the equipment. This arrangement ensures that different batches of samples will not interfere with each other when they are transferred between different modules.
  • the samples can be transferred to the next module through the driving conveyor, and then the next batch of samples can be received by the driving conveyor. In this way, different batches of samples are processed correspondingly in the corresponding modules.
  • the driving conveying device includes driving positions corresponding to the positions of the mixing module, the magnetic sorting module, and the temperature-controlled elution module, and can respectively send the mixing module and the magnetic separation module to the driving position at the driving position.
  • the selection module and the temperature-controlled elution module transmit samples and receive samples from the mixing module, the magnetic separation module and the temperature-controlled elution module.
  • Figure 3 shows such driving positions 1-3.
  • the driving vehicle can move linearly on the X axis, reaching the driving position 1, 2, 3 and the position of the sample tube respectively.
  • the bottom rotating motor realizes a 90-degree rotation of the vehicle and reaches the position of the sample tube.
  • the hook claw lifting motor realizes the fitting and disengagement of the hook claw in the sample tube slot.
  • the driving hook claw can transfer the sample tube from the oscillation module to the inside of the driving car through the front and back grabbing screw motor. After reaching the target position, the driving hook claw pushes the sample tube out of the driving car and pushes it into the destination module.
  • the pipeline nucleic acid processing equipment further includes a pipetting device arranged at the entrance of the pipeline nucleic acid processing equipment.
  • the sample to be tested is transferred to a test tube suitable for transporting in a driving conveying device through a pipetting device.
  • the car transport device can receive a sample from the pipetting device.
  • the pipetting device may include, for example, a mechanical arm and a pipetting pump, which work together to complete pipetting.
  • the sample test tube can be a standard test tube that can be purchased on the market, and the reaction cup is, for example, a 4-tube form, which can simultaneously extract samples from 4 individuals.
  • the 4-tube form of the reaction cup can also contain 4 separate pipetting tips, which can save the use of consumables.
  • the mixing module, the magnetic separation module and the temperature-controlled elution module can respectively process 1 to 4 samples at the same time.
  • each module can process up to 4 samples at the same time. This can shorten the processing time in each module and realize fast pipeline processing of samples.
  • FIG 4 is a schematic diagram of a test tube and tip placement device according to the present invention.
  • a tip placement device for placing a tip can be provided next to each test tube loaded with a sample, and the tip is placed in the tip placement device of the corresponding test tube.
  • the pipetting pump In conventional molecular pipetting workstations, the pipetting pump needs to mix the reaction system by pipetting up and down after each pipetting. Due to the high sensitivity of molecular experiments, in order to prevent cross-contamination between samples, the pipette tip can no longer process other types of samples, or pipette the same reaction liquid to other samples, even if it is in the same experimental link, the pipette pump needs to be used Replace the pipette tip before processing other samples. This not only causes a large number of pipetting tips for each test, but also because the robotic arm needs to suck the tips back and forth, the overall speed is reduced a lot.
  • the invention creatively puts the test tube and the suction head together, and runs through the entire extraction process of the clinical sample.
  • the addition of liquid can be completed by a fixed-position peristaltic pump, and the work of mixing samples is completed by the washing liquid pump combined with the pipette tip on the side of the test tube, and then this tip sucks the waste liquid and Store it on the side of the test tube again for the next step of mixing and pipetting. Since this tip only mixes and pipets a single sample, there is no pollution problem of repeated use of the tip, which not only greatly reduces the cost, but also reduces the time for the robotic arm to move back and forth due to the side-by-side prevention of the test tube and the tip. Improved system speed.
  • the combination of test tube and tip can be one, two, four, etc. multiple combinations.
  • Figure 4 shows a combination of 4 test tubes and pipette tips.
  • the tip placement device is a tube suitable for placement of the tip.
  • the tip placement device is arranged in a row with the test tube in the transverse direction of the test tube.
  • the tip placement device may also be arranged in the lateral direction of the test tube.
  • the test tube and the tip placement device can be installed together through a fixed plate and other devices.
  • the present invention creatively divides the mixing, magnetic sorting, and temperature-controlled elution required for nucleic acid extraction into three modules.
  • the modules are transferred by driving, and each module reduces the number of samples from dozens of samples at the same time to one. ⁇ 4 and so on several samples.
  • the total time of nucleic acid extraction is about 40 minutes, which is divided into three main steps: mixing, magnetic sorting, and temperature-controlled elution. Each step takes less than 10 minutes. Therefore, the traffic transfer method is used to connect each module, although the overall work throughput in a unit time is reduced, but the demand for timely processing of clinical samples from different times can be handled well.
  • an assembly line molecular diagnostic equipment including: separately arranged mixing module, magnetic separation module and temperature control elution module; Sample driving transport device; a test cartridge with pre-filled reagents, which performs an independent temperature control reaction for each sample received; an optical detection module, which detects the transferred test cartridge; and a control device for controlling the assembly line
  • the operation of the various parts of the molecular diagnostic equipment wherein the mixing module, the magnetic separation module and the temperature-controlled elution module are arranged along the moving track of the traveling conveyor, and samples are received and removed by the traveling conveyor.
  • the pipeline molecular diagnostic equipment adds a detection cartridge and an optical detection module to the pipeline nucleic acid processing equipment. That is to say, after completing the pipeline nucleic acid processing, the temperature control reaction is performed by the detection cartridge, and the detection is performed by the optical detection module.
  • the detection cassette is a disposable detection cassette with a multi-hole row tube, and has a cassette cover, which is used to open and close the row tubes in the cassette in a row, so as to independently control the processing in the detection cassette.
  • the temperature and time of the temperature control reaction are used to open and close the row tubes in the cassette in a row, so as to independently control the processing in the detection cassette. The temperature and time of the temperature control reaction.
  • FIG. 5 is a schematic diagram of a disposable cartridge according to the present invention.
  • the disposable cartridge according to the present invention can be combined with the instrument to realize the opening and closing functions of the cartridge cover, and silicone oil can be added to double isolation to prevent aerosol before amplification.
  • the lid can be opened using the principle of leverage.
  • the screw motor moves up and down to support the tail of the cover and uses the principle of leverage to open the cover; to close the cover, the screw motor moves up and down and the principle of forward and backward translation is used to move the ejector rod up and down to a specific position.
  • the lever moves down to complete the closing action.
  • the disposable cartridge pre-filled with reagents as shown in the figure can be, for example, a porous tube such as a quadruple tube or a polygonal structure such as 16 holes on four sides.
  • the common feature is that each hole is equipped with a sealing cover, and the probe is pre-packaged and then sealed. Closed and refrigerated.
  • the automatic opening and closing mechanism is used to automatically open and close the sealing cover, which effectively prevents the risk of aerosol contamination during the detection process. Since each cartridge has a different start time, the temperature control module can independently control the temperature and time of the temperature control.
  • each card box After each card box has enough temperature control time, it is transmitted to the optical detection module by the gripper or crawler, and the signal is detected separately, so as to realize the pipeline processing.
  • each part has an independent sample operation, and different steps are connected in series by a mechanical arm or the like, thereby realizing the flexibility of individual sample detection, on-the-go inspection, and improving timeliness.
  • the pipeline nucleic acid processing equipment according to the present invention can also be used for high-throughput sequencing pre-processing pipeline library construction.
  • the pre-processing for high-throughput sequencing includes DNA extraction, library construction, and DNA purification. The DNA is extracted through the pipeline method, the DNA ends are filled and connected in the cassette, and the DNA is purified through the extraction module, and finally the high-throughput sequencing pre-processing is completed.

Abstract

一种流水线核酸处理设备，包括：分开设置的混匀模块、磁力分选模块和温控洗脱模块；用于从流水线核酸处理设备的入口接收样本并进行传送的行车传送装置；和控制装置，用于控制流水线核酸处理设备中各个部分的操作；其中混匀模块、磁力分选模块和温控洗脱模块沿着行车传送装置的移动轨道设置，并通过行车传送装置接收和移出样本。一种流水线分子诊断设备，包括：分开设置的混匀模块、磁力分选模块和温控洗脱模块；用于从流水线分子诊断设备的入口接收样本并进行传送的行车传送装置；具有预填试剂的检测卡盒，对接收的每个样本进行独立的温控反应；光学检测模块，对传送来的检测卡盒进行检测；和控制装置，用于控制流水线分子诊断设备的各个部分的操作；其中混匀模块、磁力分选模块和温控洗脱模块沿着行车传送装置的移动轨道设置，并通过行车传送装置接收和移出样本。

Description

流水线核酸处理设备和流水线分子诊断设备 技术领域
本发明涉及核酸检测领域，具体而言涉及流水线核酸处理设备和流水线分子诊断设备。
背景技术
分子诊断大致分为核酸提取纯化、扩增和检测三大环节。用医院常规开展的荧光定量PCR为例，操作步骤包含核酸提取、反应配置、封膜、设置PCR程序、扩增等。目前的设计都是多个样本同时运作，便于同时处理大量的样本，做同一类型的检测，非常好地提高了分子检测的通量，可以在几个小时内处理96个甚至384个样本的快速检测。对于HBV、HPV、HIV大型筛查型的检测项目，或者第三方临检中心实验室，这样的批处理方式可以解决客户需要同时处理大量样本的需求。
然而临床在追求通量的同时也需要极高的时效性和灵活性。例如，在反应过程中，任何新增加的样本，都需要及时处理，而不是等待这一批实验结束，而往往一批实验的流程都在几个小时。或者，希望能灵活处理不同样本的不同检测需求，而不是同一类型检测项目。或者实现全流程自动化，通过扫码自动识别需要检测的内容，样本放入仪器后，全自动进行，无需人工干预。
发明内容
基于以上需求，本申请的发明人开发了基于流水线思路的分子诊断设备。例如，根据本发明的实施例，可以用流水线的方式取代批处理方式从而做到随到随检。根据其他实施例，可以通过扫描样本检测条码仪器自动判断运行的项目。
根据本发明一个方面，提供一种流水线核酸处理设备，包括：
分开设置的混匀模块、磁力分选模块和温控洗脱模块；
用于从流水线核酸处理设备的入口接收样本并进行传送的行车传送装置；和
控制装置，用于控制流水线核酸处理设备中各个部分的操作；
其中所述混匀模块、磁力分选模块和温控洗脱模块沿着所述行车传送装置的移动轨道设置，并通过行车传送装置接收和移出样本。
优选的，其中所述混匀模块、磁力分选模块和温控洗脱模块沿着行车传送装置的移动轨道顺序布置。
优选的，其中所述混匀模块布置在行车传送装置的移动轨道的上游位置。
优选的，所述行车传送装置包括分别与所述混匀模块、磁力分选模块和温控洗脱模块的位置相对应的行车位置，并且能够在所述行车位置处分别向所述混匀模块、磁力分 选模块和温控洗脱模块传送样本和从所述混匀模块、磁力分选模块和温控洗脱模块接收样本。
优选的，还包括设置在流水线核酸处理设备的入口处的移液装置。
优选的，所述行车传送装置能够从移液装置接收样本。
优选的，其中所述混匀模块、磁力分选模块和温控洗脱模块分别能够同时处理1至4个样本。
优选的，其中每个装载样本的试管旁设置有放置吸头的吸头放置装置，吸头放置在相应试管的吸头放置装置中。
根据本发明另一个方面，提供一种流水线分子诊断设备，包括：
分开设置的混匀模块、磁力分选模块和温控洗脱模块；
用于从流水线分子诊断设备的入口接收样本并进行传送的行车传送装置；
具有预填试剂的检测卡盒，对接收的样本进行独立的温控反应；
光学检测模块，对传送来的检测卡盒进行检测；和
控制装置，用于控制流水线分子诊断设备的各个部分的操作；
其中所述混匀模块、磁力分选模块和温控洗脱模块沿着所述行车传送装置的移动轨道设置，并通过行车传送装置接收和移出样本。
优选的，其中所述混匀模块、磁力分选模块和温控洗脱模块沿着行车传送装置的移动轨道顺序布置。
优选的，其中所述混匀模块布置在行车传送装置的移动轨道的上游位置。
优选的，所述行车传送装置包括分别与所述混匀模块、磁力分选模块和温控洗脱模块的位置相对应的行车位置，并且能够在所述行车位置处分别向所述混匀模块、磁力分选模块和温控洗脱模块传送样本和从所述混匀模块、磁力分选模块和温控洗脱模块接收样本。
优选的，还包括设置在流水线分子诊断设备的入口处的移液装置。
优选的，所述行车传送装置能够从移液装置接收样本。
优选的，其中所述混匀模块、磁力分选模块和温控洗脱模块分别能够同时处理1至4个样本。
优选的，其中每个装载样本的试管旁设置有放置吸头的吸头放置装置，吸头放置在相应试管的吸头放置装置中。
优选的，其中所述检测卡盒是具有多孔排管的一次性检测卡盒，具有能够开启和关闭的卡盒盖子，并能够控制一次性检测卡盒内接收的样本进行的温控反应的温度和时间。
根据本发明的优选实施例，每个样本的检测内容可以灵活组合，兼顾较高通量、灵活性和自动化。
附图说明
通过以下详细的描述并结合附图将更充分地理解本发明，其中相似的元件以相似的方式编号，其中：
图1是根据本发明一个实施例的流水线分子诊断设备的示意图；
图2是图1的流水线分子诊断设备的结构示意图；
图3是根据本发明一个实施例的流水线核酸处理设备的示意图；
图4是根据本发明的试管和吸头放置装置的示意图；
图5是根据本发明的一次性卡盒的示意图；和
图6A和6B是根据本发明的行车传送装置的工作示意图。
具体实施方式
下面通过实施例，并结合附图，对本发明的技术方案作进一步详细的说明，但本发明不限于下面的实施例。
分子诊断基本包括核酸提取、反应配置、检测等几个环节。整个流程过程比较繁琐，基本是手工使用为主。以常见的荧光定量PCR为例，核酸提取40分钟，反应体系配置30分钟，荧光定量PCR两个小时，加上每个步骤中间的手工操作和等待时间，最后整体试验全流程往往在4-5个小时。要实现自动化基本以TECAN、Hamilton等公司为代表的移液工作站为主，其大致结构原理基本是在一个水平台面加上4、8或96通道的移液泵，在底部放置存放液体、样本的耗材。样本和各种试剂在耗材内通过移液泵混合、反应。基本的耗材有24、48、96和384孔板。废液和废弃吸头由驱动移液泵的机械臂移动到废置位置废弃。优点是一次性可以处理大量的样本(24、48、96等数量样本)执行同样类型的检测，但难以应对不同时间出现的样本检测需求。为了不引起样本之间的交叉污染，每次移液混合以后，移液泵都需要更换移液吸头。一个简单的提取过程就有加样，加裂解液，吸废液，加清洗液，吸废液，加清洗液，吸废液，加清洗液，吸废液，加洗脱液洗脱等十个步骤，洗液泵需要来回样本耗材处和移液吸头处10次，需要10个移液吸头。每次更换以1秒计算，提取过程洗液泵就需要移动10个来回，共10分钟，占比核酸提取整个操作过程40％时间。移液平台越大，整个过程时间更长。
为了解决这些问题，本发明把常规批处理的分子诊断方式改成流水线形式。
具体地，根据本发明一个实施例，可以将核酸提取过程拆分成混匀模块、磁力分选模块、温控洗脱模块三个模块。三个模块之间，通过行车的方式来移动样本，而不是常规用一个移液泵处理。这样虽然降低了通量，但是通过把核酸提取的整体时间打散，每个单独步骤的时间约几分钟。这样基本可以做到随时增加新的样本，缩短了每一个单独样本的临床结果交付时间。
提取后的样本通过移液进入预填检测探针的独立卡盒。卡盒可以是多联管、多边形等结构，顶部具备自动开盖功能，在卡盒内完成独立温控反应。最后由光学检测模块独立检测信号。
通过上述方式，实现了样本在整个流程中一直在不同工位上流动，实现样本进结果出，无需人工干预，全流程一体化，真正流水线检测模式。
图1是根据本发明一个实施例的流水线分子诊断设备的示意图；图2是图1的流水线分子诊断设备的结构示意图；图3是根据本发明一个实施例的流水线核酸处理设备的示意图。
如图3所示，根据本发明一个实施例，提供一种流水线核酸处理设备，包括：分开设置的混匀模块、磁力分选模块和温控洗脱模块；用于从流水线核酸处理设备的入口接收样本并进行传送的行车传送装置；和控制装置，用于控制流水线核酸处理设备中各个部分的操作；其中所述混匀模块、磁力分选模块和温控洗脱模块沿着所述行车传送装置的移动轨道设置，并通过行车传送装置接收和移出样本。
如图3、图6A和6B所示，根据本发明的行车传送装置可以包括用于进行传送的行车和行车在上面移动的移动轨道。行车可以带有钩爪以抓取和卸载反应杯或样本管，或其他样品装载容器。例如，可以利用丝杆电机通过钩爪将反应杯拖入行车内，行车旋转至便于在移动轨道上进行移动的移动位，利用带轮平移功能将行车移动至下一个工位或行车位置，钩爪和丝杆电机可以完成反应杯的卸载工作。
行车传送装置的移动轨道可以基本是直线的，或者也可以是其他线性形状轨道。所述混匀模块、磁力分选模块和温控洗脱模块沿着行车传送装置的移动轨道顺序布置。其中所述混匀模块布置在行车传送装置的移动轨道的上游位置。上游位置通常是指更靠近设备入口的位置。这样的布置方式使得不同批次的样本在不同模块之间传送时不会彼此发生干涉。当处理完成当前批次的样本后，可以通过行车传送装置将样本传送到下一个模块，然后继续通过行车传送装置接收下一个批次的样本。这样，不同批次的样本在相应的模块中进行对应的处理。
所述行车传送装置包括分别与所述混匀模块、磁力分选模块和温控洗脱模块的位置相对应的行车位置，并且能够在所述行车位置处分别向所述混匀模块、磁力分选模块和温控洗脱模块传送样本和从所述混匀模块、磁力分选模块和温控洗脱模块接收样本。图3示出了这样的行车位置1-3。
如图3所示，行车在X轴可以直线运动，分别到达行车位置1,2,3以及样本管位置。通过底部旋转电机实现行车90度旋转，到达样本管位置。由钩爪升降电机实现钩爪在样本管卡槽的套合与脱离。行车钩爪可以通过前后抓取丝杆电机实现样本管从振荡模块传送到行车内部，到达目的位置后，再由行车钩爪将样本管推出行车，推至目的模块内部。
流水线核酸处理设备还包括设置在流水线核酸处理设备的入口处的移液装置。通过移液装置将送检的样本转移到适于在行车传送装置进行传送的试管中。
所述行车传送装置能够从移液装置接收样本。
根据本发明的移液装置例如可以包括机械臂和移液泵，其联合运行完成移液。样本试管可以采用市场上能够购买的标准试管，反应杯例如采用4联管形式，可同时满足4 个人样本同时提取。4联管形式的反应杯还可以包含4个单独的移液吸头，可以节约耗材的使用。
优选的，其中所述混匀模块、磁力分选模块和温控洗脱模块分别能够同时处理1至4个样本。根据优先实施方式，每个模块最多可以同时处理4个样本。这样可以缩短在每个模块的处理时间，实现样本的快速流水线处理。
图4是根据本发明的试管和吸头放置装置的示意图。如图4所示，每个装载样本的试管旁可以设置有放置吸头的吸头放置装置，吸头放置在相应试管的吸头放置装置中。
在常规分子移液工作站中，每次移液结束后移液泵需要通过上下吸打来混匀反应体系。由于分子实验灵敏度非常高，为了防止样本间交叉污染因此该移液吸头不能再处理其他类型样本，或者移液同样反应液体到其他样本，即使还在同样的实验环节，移液泵也需要去更换移液吸头再处理其他样本。这样不仅造成了每次试验需要大量的移液吸头，同时由于机械臂需要来回吸打枪头，整体速度降低很多。
对于每一个单独样本来说同样的移液吸头反复使用不会有样本间污染问题。常规的移液工作站所有耗材都只是存放液体，没有暂存样本吸头的位置。如果要反复使用吸头，机械臂也需要在移液位置和吸头原始存放位置之间来回移动，不光速度会减慢，洗液泵带着有残留液体的吸头在工作站内来回移动，也增加了污染其他样本的可能。
本发明创造性的把试验管和吸头放在一起，随着临床样本整个提取流程全部贯串。这样每一个样本核酸提取过程中，添加液体可以由固定位置的蠕动泵完成，混合样本的工作由洗液泵结合在试管边上的移液吸头完成，再由这个吸头吸去废液并再次存放在试管边上以备下一步混合移液工作。由于这个吸头都只混合移液一个单独样本，因此不存在反复使用吸头的污染问题，不仅大大降低了费用，同时由于试管和吸头并排防止，减少了机械臂来回移动吸打的时间，提高了系统速度。试管和吸头组合可以是一个、两个、四个等多个组合。图4示出的是4个试管和吸头组合的情形。
在图4中，吸头放置装置是适于放置吸头的管。在图4中，吸头放置装置设置在试管的横向方向与试管形成一排。在其他实施方式中，吸头放置装置也可以设置在试管的侧向方向上。试管和吸头放置装置可以通过固定盘等装置安装在一起。
从下表能看到在核酸提取环节，使用常规移液工作站，8联泵处理8个样本，也就是每一个移液泵一个样本，整个过程需要6个移液吸头；如果要8联泵处理大于8个样本，就需要反复更换移液吸头，整个过程至少需要10个移液吸头。而本发明大部分环节只需要一个吸头，最后由于要保证洗脱样本的洁净，不再重复使用，整个过程仅需两个吸头，最高可以降低80％的移液吸头使用量，并减少了大量机械臂来回运动的时间，避免了不必要的污染。
表1 所需移液吸头对比表
Figure PCTCN2021090890-appb-000001
常规核酸提取为批处理模式，难以应对在不同时间出现的样本及不同样本不用检测项目的需求。本发明创造性地把核酸提取所需要的混匀、磁力分选、温控洗脱分为三个模块，模块之间用行车来进行传递，每个模块从原来同时处理几十个样本减少到1～4等数个样本。核酸提取总时间约为40分钟，分别分为混匀、磁力分选、温控洗脱三个主要步骤，每个步骤单独时间小于10分钟。因此用行车传递方式来连接各个模块，虽然降低了在一个单位时间内整体作业通量，但是可以很好地处理不同时间来的临床样本需要及时处理的需求。
如图1和2所示，根据本发明，提供一种流水线分子诊断设备，包括：分开设置的混匀模块、磁力分选模块和温控洗脱模块；用于从流水线分子诊断设备的入口接收样本的行车传送装置；具有预填试剂的检测卡盒，对接收的每个样本进行独立的温控反应；光学检测模块，对传送来的检测卡盒进行检测；和控制装置，用于控制流水线分子诊断设备的各个部分的操作；其中所述混匀模块、磁力分选模块和温控洗脱模块沿着所述行车传送装置的移动轨道设置，并通过行车传送装置接收和移出样本。
关于图3中流水线核酸处理设备的各个部件的说明同样适用于图1和图2中的流水线分子诊断设备，此处不再赘述。
实际上，流水线分子诊断设备是在流水线核酸处理设备的基础上增加了检测卡盒和光学检测模块。也就是说，在完成流水线核酸处理后，通过检测卡盒进行温控反应，并通过光学检测模块进行检测。
优选的，其中所述检测卡盒是具有多孔联排管的一次性检测卡盒，具有卡盒盖子，用于联排地开关卡盒中的联排管，从而独立控制检测卡盒内进行的温控反应的温度和时间。
图5是根据本发明的一次性卡盒的示意图。根据本发明的一次性卡盒配合仪器可以实现卡盒盖子开启和关闭功能，在扩增前可加入硅油双重隔绝防止气溶胶。例如，可以利用杠杆原理打开盖子。丝杆电机上下移动顶住盖子尾部利用杠杆原理顶开盖子；关闭盖子采用丝杆电机上下运动和前后平移的原理，将顶杆上下移动到特定位置，前后移动电机推动顶杆从而推动盖子，顶杆向下移动，完成关盖动作。如图所示预填试剂的一次 性卡盒例如可以为多孔排管比如四联管或多边形结构比如四边16孔，共同特点是每个孔都各带一个密封盖，探针预封装后密封盖闭合冷藏。在设备中按流程需求，使用自动开闭盖机构自动开启和关闭密封盖，有效防止检测过程中气溶胶污染的风险。每一个卡盒由于开始时间不同，在温控模块都可以独立控制温控温度和时间。
每一个卡盒有足够温控时间后，由夹爪或履带传递到光学检测模块，单独检测信号，从而实现流水线处理。
根据本发明，每一个部分都独立的样本操作，不同的步骤之间通过机械臂等串联起来，从而实现了单独样本检测的灵活性、随到随检，并提高了时效性。
此外，根据本发明的流水线核酸处理设备也可以配合用于高通量测序前处理流水线文库构建。高通量测序前处理包括DNA提取、文库构建、DNA纯化。通过流水线方式提取DNA、在卡盒内完成DNA末端补平、连接，同时通过提取模块进行DNA纯化，最终完成高通量测序前处理。
本发明的实施方式并不限于上述实施例所述，在不偏离本发明的精神和范围的情况下，本领域普通技术人员可以在形式和细节上对本发明做出各种改变和改进，而这些均被认为落入了本发明的保护范围。

Claims (17)

  1. 一种流水线核酸处理设备，包括：
    分开设置的混匀模块、磁力分选模块和温控洗脱模块；
    用于从流水线核酸处理设备的入口接收样本并进行传送的行车传送装置；和
    控制装置，用于控制流水线核酸处理设备中各个部分的操作；
    其中所述混匀模块、磁力分选模块和温控洗脱模块沿着所述行车传送装置的移动轨道设置，并通过行车传送装置接收和移出样本。
  2. 根据权利要求1所述的流水线核酸处理设备，其中所述混匀模块、磁力分选模块和温控洗脱模块沿着行车传送装置的移动轨道顺序布置。
  3. 根据权利要求2所述的流水线核酸处理设备，其中所述混匀模块布置在行车传送装置的移动轨道的上游位置。
  4. 根据权利要求1-3中任一项所述的流水线核酸处理设备，所述行车传送装置包括分别与所述混匀模块、磁力分选模块和温控洗脱模块的位置相对应的行车位置，并且能够在所述行车位置处分别向所述混匀模块、磁力分选模块和温控洗脱模块传送样本和从所述混匀模块、磁力分选模块和温控洗脱模块接收样本。
  5. 根据权利要求1-4中任一项所述的流水线核酸处理设备，还包括设置在流水线核酸处理设备的入口处的移液装置。
  6. 根据权利要求5所述的流水线核酸处理设备，所述行车传送装置能够从移液装置接收样本。
  7. 根据权利要求1-6中任一项所述的流水线核酸处理设备，其中所述混匀模块、磁力分选模块和温控洗脱模块分别能够同时处理1至4个样本。
  8. 根据权利要求1-7中任一项所述的流水线核酸处理设备，其中每个装载样本的试管旁设置有放置吸头的吸头放置装置，吸头放置在相应试管的吸头放置装置中。
  9. 一种流水线分子诊断设备，包括：
    分开设置的混匀模块、磁力分选模块和温控洗脱模块；
    用于从流水线分子诊断设备的入口接收样本并进行传送的行车传送装置；
    具有预填试剂的检测卡盒，对接收的样本进行独立的温控反应；
    光学检测模块，对传送来的检测卡盒进行检测；和
    控制装置，用于控制流水线分子诊断设备的各个部分的操作；
    其中所述混匀模块、磁力分选模块和温控洗脱模块沿着所述行车传送装置的移动轨道设置，并通过行车传送装置接收和移出样本。
  10. 根据权利要求9所述的流水线分子诊断设备，其中所述混匀模块、磁力分选模块和温控洗脱模块沿着行车传送装置的移动轨道顺序布置。
  11. 根据权利要求10所述的流水线分子诊断设备，其中所述混匀模块布置在行车传送装置的移动轨道的上游位置。
  12. 根据权利要求9-11中任一项所述的流水线分子诊断设备，所述行车传送装置包括分别与所述混匀模块、磁力分选模块和温控洗脱模块的位置相对应的行车位置，并且能够在所述行车位置处分别向所述混匀模块、磁力分选模块和温控洗脱模块传送样本和从所述混匀模块、磁力分选模块和温控洗脱模块接收样本。
  13. 根据权利要求9-12中任一项所述的流水线分子诊断设备，还包括设置在流水线分子诊断设备的入口处的移液装置。
  14. 根据权利要求13所述的流水线分子诊断设备，所述行车传送装置能够从移液装置接收样本。
  15. 根据权利要求9-14中任一项所述的流水线分子诊断设备，其中所述混匀模块、磁力分选模块和温控洗脱模块分别能够同时处理1至4个样本。
  16. 根据权利要求9-15中任一项所述的流水线分子诊断设备，其中每个装载样本的试管旁设置有放置吸头的吸头放置装置，吸头放置在相应试管的吸头放置装置中。
  17. 根据权利要求9-16中任一项所述的流水线分子诊断设备，其中所述检测卡盒是具有多孔排管的一次性检测卡盒，具有能够开启和关闭的卡盒盖子，并能够控制一次性检测卡盒内接收的样本进行的温控反应的温度和时间。
PCT/CN2021/090890 2020-04-30 2021-04-29 流水线核酸处理设备和流水线分子诊断设备 WO2021219067A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202010366043 2020-04-30
CN202010366043.7 2020-04-30

Publications (1)

Publication Number Publication Date
WO2021219067A1 true WO2021219067A1 (zh) 2021-11-04

Family

ID=78374069

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/090890 WO2021219067A1 (zh) 2020-04-30 2021-04-29 流水线核酸处理设备和流水线分子诊断设备

Country Status (1)

Country Link
WO (1) WO2021219067A1 (zh)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114164086A (zh) * 2021-12-07 2022-03-11 江苏领坤生物科技有限公司 全自动移液工作站用的移液平台
CN114276896A (zh) * 2021-12-22 2022-04-05 成都瀚辰光翼科技有限责任公司 一种自动化核酸提取系统、方法及存储介质
CN115747031A (zh) * 2022-11-17 2023-03-07 北京昌平实验室 病原体核酸分析设备
CN116676183A (zh) * 2023-07-31 2023-09-01 四川徕伯益自动化技术有限公司 一种分子诊断系统、分子诊断方法

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090117620A1 (en) * 2007-11-05 2009-05-07 Abbott Laboratories Automated analyzer for clinical laboratory
CN101613663A (zh) * 2009-07-28 2009-12-30 呼吸疾病国家重点实验室 全自动串行式荧光pcr检测系统
CN105188938A (zh) * 2013-03-14 2015-12-23 简·探针公司 诊断系统和方法
CN106404508A (zh) * 2016-11-23 2017-02-15 杭州杰毅麦特医疗器械有限公司 一种核酸检测前处理自动化处理装置
CN106754339A (zh) * 2016-12-14 2017-05-31 杭州杰毅麦特医疗器械有限公司 核酸检测前处理自动化处理装置
CN108220155A (zh) * 2016-12-21 2018-06-29 康代思锐公司 用于分子诊断的系统和方法
CN109337807A (zh) * 2018-09-30 2019-02-15 苏州百源基因技术有限公司 Pcr流水线
CN110564607A (zh) * 2019-09-11 2019-12-13 长春技特生物技术有限公司 全自动核酸提取扩增微流控芯片动态定量检测一体化装置

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090117620A1 (en) * 2007-11-05 2009-05-07 Abbott Laboratories Automated analyzer for clinical laboratory
CN101613663A (zh) * 2009-07-28 2009-12-30 呼吸疾病国家重点实验室 全自动串行式荧光pcr检测系统
CN105188938A (zh) * 2013-03-14 2015-12-23 简·探针公司 诊断系统和方法
CN106404508A (zh) * 2016-11-23 2017-02-15 杭州杰毅麦特医疗器械有限公司 一种核酸检测前处理自动化处理装置
CN106754339A (zh) * 2016-12-14 2017-05-31 杭州杰毅麦特医疗器械有限公司 核酸检测前处理自动化处理装置
CN108220155A (zh) * 2016-12-21 2018-06-29 康代思锐公司 用于分子诊断的系统和方法
CN109337807A (zh) * 2018-09-30 2019-02-15 苏州百源基因技术有限公司 Pcr流水线
CN110564607A (zh) * 2019-09-11 2019-12-13 长春技特生物技术有限公司 全自动核酸提取扩增微流控芯片动态定量检测一体化装置

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114164086A (zh) * 2021-12-07 2022-03-11 江苏领坤生物科技有限公司 全自动移液工作站用的移液平台
CN114276896A (zh) * 2021-12-22 2022-04-05 成都瀚辰光翼科技有限责任公司 一种自动化核酸提取系统、方法及存储介质
CN114276896B (zh) * 2021-12-22 2023-12-29 成都瀚辰光翼科技有限责任公司 一种自动化核酸提取方法及存储介质
CN115747031A (zh) * 2022-11-17 2023-03-07 北京昌平实验室 病原体核酸分析设备
CN116676183A (zh) * 2023-07-31 2023-09-01 四川徕伯益自动化技术有限公司 一种分子诊断系统、分子诊断方法
CN116676183B (zh) * 2023-07-31 2023-11-10 四川徕伯益自动化技术有限公司 一种分子诊断系统

Similar Documents

Publication Publication Date Title
WO2021219067A1 (zh) 流水线核酸处理设备和流水线分子诊断设备
JP7446378B2 (ja) 自動分析のためのシステム及び方法
US20210302456A1 (en) Sample analysis device
US8282895B2 (en) Reagent cabinet system
JP4532264B2 (ja) 自動システム及び自動処理方法並びに核酸自動抽出方法
JP5038004B2 (ja) 分析装置
US10458998B2 (en) Device for processing samples
EP2047282B1 (en) Device for processing samples
CN109313207B (zh) 试样处理测量系统
CN103635809B (zh) 自动分析系统
CN116396850A (zh) 一种全自动核酸样品处理系统
CN111575172A (zh) 基因检测系统
EP1615037A1 (en) An apparatus for liquid handling with multiple transfer tools
CN212610645U (zh) 基因检测系统
JP2005274153A (ja) 検体分析機のローダー装置及びアンローダー装置
JP5848559B2 (ja) 自動分析装置
CN220961555U (zh) 一种试样分配装置及样本分析仪
JP4607677B2 (ja) 自動分析装置の容器搬送機構
US20220168733A1 (en) Device and method for the extraction of nucleic acids
CN117242351A (zh) 用于汇集样本以进行高通量分析的系统和方法
CN117511722A (zh) 分子诊断分析仪和分子诊断方法
JP2012189404A (ja) 臨床検査用分析装置

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21797633

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21797633

Country of ref document: EP

Kind code of ref document: A1

122 Ep: pct application non-entry in european phase

Ref document number: 21797633

Country of ref document: EP

Kind code of ref document: A1