TW202321431A - Nucleic acid processing device and sample pre-processing module thereof - Google Patents

Abstract

A nucleic acid processing device includes a sample pre-processing module and a nucleic acid purification module. The sample pre-processing module includes a sample chamber, plural liquid storage chambers, a mixing chamber, plural communication channels, and a first connection structure. The sample chamber has a sample loading port for adding a sample into the sample chamber. The plural liquid storage chambers are configured to store plural sample pre-processing reagents, which at least include a lysis buffer and a proteinase solution. The mixing chamber is provided for mixing the sample and the sample pre-processing reagents. The plural communication channels are configured to transmit the sample and the sample pre-processing reagents to the mixing chamber. The first connection structure is detachably connected with a second connection structure of the nucleic acid purification module, so as to transmit the pre-processed sample to the nucleic acid purification module.

Description

Nucleic acid processing device and its sample pretreatment module

This case is about a nucleic acid processing device and its sample pre-processing module, especially a nucleic acid processing device and its detachable sample pre-processing module.

As the global new crown virus epidemic continues to rage and intensify, public health is a difficult challenge that countries around the world are facing today. In the case of the rapid spread of infectious diseases, effective prevention and control strategies, such as rapid detection and confirmation of suspected cases, early isolation or confirmation of treatment, are the goals of the joint efforts of the global detection medicine institutes. Therefore, the development of "instant and on-site detection methods" for infectious diseases is one of the urgent issues. At present, although rapid screening immunization is mostly used for detection, the 30-minute preliminary screening is only suitable for detecting high-concentration antigen/antibody samples, and is only suitable for a small number of infectious diseases, so it has its limitations. The molecular detection method can provide high-accuracy analysis. However, due to the limitation of "the demand for large and expensive machines" and "the need for professionals to operate complicated procedures", this detection technology can only It can only be tested in biochemical research and development centers or clinical analysis laboratories with professional and technical personnel.

The development of "chip laboratory" has gradually matured in recent years. Its core concept is to miniaturize the functions of different testing equipment and integrate them on the same testing platform to realize the so-called "point of care (POC)" in vitro diagnosis (In Vitro Diagnostics, IVD), which has the advantages of "small system size", "high accuracy", "rapid response" and "instant diagnosis". Based on microfluidic technology, it is now possible to (1) extract and purify pathogen nucleic acid (DNA or RNA), and (2) carry out many expensive instruments and complicated operating procedures required by traditional nucleic acid detection steps such as nucleic acid amplification and detection , integrated on a "biological chip cassette", and combined with the development of a fully automatic nucleic acid analyzer, it is hoped to improve the defects of the diagnosis of infectious diseases today, provide the goal of real-time on-site detection, and then create the vision of the well-being of all human beings.

In response to the sample collection required for testing, such as different sample shapes or different collection methods, it is usually necessary to pre-process the sample before sending it to the nucleic acid analyzer for nucleic acid extraction, amplification, and detection. Therefore, how to simplify the cumbersome steps required for sample pretreatment to make sample collection, storage and transportation more convenient is an urgent problem to be overcome.

One purpose of this case is to provide a sample pretreatment module, which can replace the manual operation before the sample is sent to the nucleic acid analyzer, simplify the cumbersome steps required for sample pretreatment, and make sample collection, storage and transportation more convenient.

Another object of this case is to provide a nucleic acid processing device, including a detachable sample pretreatment module, which can be connected to the nucleic acid purification module, and is used to transport the pretreated sample to the nucleic acid purification module for subsequent Nucleic acid extraction, amplification and detection.

In order to achieve the above purpose, this case provides a nucleic acid processing device, including a sample pretreatment module and a nucleic acid purification module, wherein the sample pretreatment module includes: a sample tank with a sample port for adding samples to the sample tank; The liquid storage tank is configured to store a plurality of pre-treatment reagents, and the pre-treatment reagents include at least lysate and protease; the mixing tank is configured to mix samples and pre-treatment reagents; a plurality of communication pipes are configured to transport samples and pre-treatment reagents to the mixing tank; and the first connection structure is detachably connected to the second connection structure of the nucleic acid purification module, and is configured to transport the pre-treated sample to the nucleic acid purification module.

In one embodiment, the sample tank contains a sample preservation solution, such as a virus transport preservation solution, a universal transport preservation solution, a bacteria transport preservation solution, or a tissue cell preservation solution.

In one embodiment, the pretreatment reagent further includes a nucleic acid precipitation aid, such as carrier RNA, ethanol, or glycogen.

In one embodiment, the first connection structure is a convex portion, and the second connection structure is a concave portion.

In one embodiment, the nucleic acid processing device further includes a cover covering the sample pretreatment module and the nucleic acid purification module. The cover includes a bridge pneumatic valve, which is configured to drive the sample to flow from the sample pretreatment module to the nucleic acid purification module.

In one embodiment, the sample pretreatment module is connected with an external cracking module. The external cracking module can be an ultrasonic cracking module, a heating cracking module, an optically driven cracking module, a magnetically driven particle grinding module, or a mechanical grinding module.

In one embodiment, the nucleic acid processing device further includes a nucleic acid amplification module connected to the nucleic acid purification module and the external active energy supply module. The external active energy supply module can be an ultrasonic hybrid module, a magnetically driven particle disturbance module, an optically driven induced heating module, or an inductive heating module.

In one embodiment, the sample is a swab, urine, blood, sputum, feces, or saliva.

In order to achieve the above purpose, this case further provides a sample pre-treatment module, including: a sample tank with a sample injection port for adding samples to the sample tank; a plurality of liquid storage tanks, which are structured to store a plurality of pre-treatment reagents, pre-treatment The reagents include at least lysate and protease; the mixing tank is configured to mix the sample and pre-treatment reagents; a plurality of communication pipes are configured to transport the sample and pre-treatment reagents to the mixing tank; and the output structure is configured to be pre-treated The sample output.

Some embodiments embodying the features and advantages of the present application will be described in detail in the description in the following paragraphs. It should be understood that the present case can have various changes in different aspects without departing from the scope of the present case, and the descriptions and drawings therein are used for illustration in nature rather than limiting the present case.

This case mainly provides a sample pretreatment module, including multiple tanks and pretreatment reagents, which can replace the manual operation before the sample is sent to the nucleic acid analyzer, and is convenient for sample collection, storage and transportation. In particular, the sample pretreatment module in this case is detachably connected to the nucleic acid purification module, which is used to transport the pretreated samples to the nucleic acid purification module for subsequent nucleic acid extraction, amplification and detection. On the other hand, this case also provides a nucleic acid processing device, including a sample pretreatment module and a nucleic acid purification module, wherein the sample pretreatment module and the nucleic acid purification module have a corresponding connection structure, so that the sample pretreatment module can be It is detachably connected with the nucleic acid purification module.

Figure 1 shows a schematic diagram of the assembly of the nucleic acid processing device of this case, and Figure 2 shows a schematic diagram of the nucleic acid processing device with the lid separated. The nucleic acid processing device mainly includes a sample pretreatment module 1 and a nucleic acid purification module 2. The sample pretreatment module 1 is used to perform pretreatment such as initial extraction on the sample, and is connected to the nucleic acid purification module 2 to convert the pretreated The sample is sent to the nucleic acid purification module 2 for purification to complete the nucleic acid extraction and facilitate the subsequent amplification and detection processes.

In one embodiment, the nucleic acid processing device further includes a cover 3, which covers the sample pretreatment module 1 and the nucleic acid purification module 2, and has slots corresponding to the sample pretreatment module 1 and the nucleic acid purification module 2 The pneumatic opening 31 and the infusion pipeline 32 of the body. The pneumatic opening 31 can be connected with a piezoelectric micropump to provide a gas pressure source, and the solenoid valve controls the switch of gas input into each tank to drive the flow of fluid between the tanks.

In one embodiment, the nucleic acid processing device further includes a nucleic acid amplification module 4, which is arranged below the nucleic acid purification module 2 and connected to the nucleic acid purification module 2, for collecting the extracted nucleic acid solution and performing subsequent nucleic acid amplification. increase and detection. In one embodiment, the nucleic acid amplification module 4 includes a collection tube 41 , which can be an eppendorf tube, but not limited thereto.

Figure 3 shows an exploded view of the nucleic acid processing device. The sample pretreatment module 1 includes a housing 11 and a tank structure 12 covered therein. The nucleic acid purification module 2 includes a shell 21 and a tank structure 22 covered therein, a reaction column 23 and a distribution component 24 . FIG. 4 and FIG. 5 further show exploded views of the tank structure 12 of the sample pretreatment module 1 and the tank structure 22 of the nucleic acid purification module 2 at different viewing angles. The tank structure 12 of the sample pretreatment module 1 includes a sample tank 121 , a plurality of liquid storage tanks 122 , a mixing tank 123 , a plurality of communication pipes 124 , and a first connection structure 125 . The sample tank 121 has a sample injection port 1211 for adding a sample into the sample tank 121 . A plurality of liquid storage tanks 122 are configured to store a plurality of pretreatment reagents. The mixing tank 123 is configured for mixing the sample and the pretreatment reagent. A plurality of communication pipes 124 are configured to transport samples and pretreatment reagents to the mixing tank 123 . The first connection structure 125 is detachably connected to the nucleic acid purification module 2, and is configured to transport the pretreated sample to the nucleic acid purification module 2, so as to facilitate a complete nucleic acid extraction process.

In one embodiment, the sample pretreatment module 1 has a sample preservation solution for stably preserving the sample during transportation. The sample storage medium can be selected according to the type of sample, such as but not limited to viral transport storage medium (viral transport medium, VTM), universal transport storage medium (universal transport medium, UTM), bacterial transport storage solution (bacterial transport medium, BTM), Or tissue cell preservation solution.

In one embodiment, the sample preservation solution can be pre-stored in the sample tank 121 for direct insertion of the swab for sampling. In another embodiment, the sample preservation solution has been stored in the sampling tube, so the sample preservation solution can be moved from the sampling tube to the sample tank 121 .

Sample pretreatment is mainly to lyse the sample before nucleic acid extraction, so as to completely destroy the cell wall and cell membrane of solid tissues and cells, thereby releasing the nucleic acid in the sample, including DNA and RNA, so that the subsequent extraction steps are easier and more efficient. efficiency. Pretreatment reagents include at least lysate and protease. The lysate disrupts cells or bacteria to lyse cells and release nucleic acids. Proteases break down proteins and separate DNA or RNA from proteins.

In one embodiment, the pretreatment reagent further includes a nucleic acid precipitation aid, such as but not limited to carrier RNA (Carrier RNA), ethanol, or glycogen. Nucleic acid precipitation aid is helpful for the effective precipitation of DNA or RNA, which can improve the recovery rate of nucleic acid.

The sample after preliminary extraction by the sample pretreatment module 1 will then be sent to the nucleic acid purification module 2 for purification to complete the nucleic acid extraction. Therefore, the sample pretreatment module 1 and the nucleic acid purification module 2 have corresponding connection structures, including the first connection structure 125 on the sample pretreatment module 1 and the second connection structure 25 on the nucleic acid purification module 2 . In one embodiment, the first connection structure 125 is a convex portion, and has a liquid delivery pipe 1251 communicating with the mixing tank 123; and the second connection structure 25 is a concave portion, which can be directly connected to the first connection structure 125, so that The sample after preliminary extraction can be sent to the nucleic acid purification module 2 from the sample pretreatment module 1 . Certainly, the connection form of the sample pretreatment module 1 and the nucleic acid purification module 2 is not limited to the above-mentioned embodiment of the convex part and the concave part, and any other connection structures that can be docked and detached are applicable to this case.

In one embodiment, as shown in Figures 1 and 2, the cover body 3 covers the sample pretreatment module 1 and the nucleic acid purification module 2, and includes a top surface and four side walls, wherein the four side walls It can be attached to the shell of the sample pretreatment module 1 and the nucleic acid purification module 2 after assembly, so the cover body 3 bridges and fits the sample pretreatment module 1 and the nucleic acid purification module 2, and also includes a bridge pneumatic The valve 33 is configured to drive the sample to flow from the sample pretreatment module 1 to the nucleic acid purification module 2 .

The tank structure 22 of the nucleic acid purification module 2 has multiple tanks and tank communication pipes. A plurality of tanks include a sample tank, a liquid storage tank, and a waste liquid tank, wherein the sample tank is configured to receive samples from the sample pretreatment module 1; Buffer solution, etc.; the waste liquid tank structure is used to store the waste liquid after the reaction. The reaction column 23 is disposed in the tank structure 22 and has a nucleic acid capture material. The tank communication pipes are constructed to transport fluid between the tanks and the reaction column 23 . The sample from the sample pretreatment module 1 will be sent to the reaction column 23, so that the nucleic acid is combined with the nucleic acid capture material in the reaction column 23, and then washed with the washing buffer to remove the residual protein and cells Finally, the nucleic acid is eluted and separated by the elution buffer, and the extracted nucleic acid solution is sent to the collection tube 41 of the nucleic acid amplification module 4 by the distributing component 24 .

In one embodiment, as shown in FIG. 6 , in order to increase the efficiency of cell destruction or bacteria destruction, the sample pretreatment module 1 of this case can be connected with an external lysis module 5 to improve the lysis efficiency. For example, the external cracking module 5 can be, but not limited to, an ultrasonic cracking module, a heating cracking module, an optically driven cracking module, a magnetically driven particle grinding module, or a mechanical grinding module.

In one embodiment, as shown in FIG. 6 , the nucleic acid amplification module 4 can be connected to an external active energy supply module 6 to improve reagent efficiency. For example, the external active energy supply module 6 can be, but not limited to, an ultrasonic mixing module, a magnetically driven particle disturbance module, a light-driven induced heating module, or an inductive heating module.

In one embodiment, the nucleic acid amplification module 4 can be connected with a nucleic acid detection device to perform a complete nucleic acid analysis process. For example, after the sample pretreatment module 1 is joined with the nucleic acid purification module 2 and the nucleic acid amplification module 4, it can be put into an automatic nucleic acid analyzer for sample pretreatment, nucleic acid extraction, nucleic acid amplification, and nucleic acid analysis. Detection and other processes, and obtain analysis results.

In one embodiment, the applicable specimen samples of the sample preprocessing module 1 include but not limited to swabs, urine, blood, sputum, feces, and saliva, etc., and the samples contained in the sample preprocessing module 1 Pretreatment reagents can adjust the reagent formula for different samples.

According to the idea of this case, the initial sample after sampling, such as a sample from a swab or a sampling tube, will first be sent to the sample tank 121 of the sample pretreatment module 1 through the sample inlet 1211, and since there is a sample in the sample tank 121 Preservative solution, so samples can be stored stably during shipping. Furthermore, since the pretreatment reagents, such as lysate, protease, and nucleic acid precipitation aid, have been pre-stored in the liquid storage tank 122 of the sample pretreatment module 1, there is no need to manually add them, which can simplify the sample pretreatment process. No cumbersome steps are required, making sample collection, storage and transportation more convenient. Since the sample pre-processing module 1 is a detachable structure, it can be directly used for the collection of the first line. After the collection is completed, the sample pre-processing module 1 can be transported to the laboratory, and the sample pre-processing module will be removed by the personnel of the laboratory. 1 is connected with the nucleic acid purification module 2, and then sent to the automatic nucleic acid analyzer for subsequent sample pretreatment, nucleic acid extraction, nucleic acid amplification, and nucleic acid detection processes. That is to say, through the fluid control module of the automatic nucleic acid analyzer, the sample and the pretreatment reagent can be mixed in the sample pretreatment module 1 for the primary extraction of the lysate, and then the sample after the primary extraction is sent to the nucleic acid purification module 2 In the process, the nucleic acid is further purified and extracted, and then the nucleic acid is amplified and detected.

Furthermore, since different specimen samples require different formulations of pretreatment reagents, the sample pretreatment module 1 is designed as a detachable cartridge, so that different pretreatment reagents can be configured for different samples, and It can cooperate with the collection, storage and transportation of the front line. Compared with fully automatic analyzers that change the cartridge design in response to different sample needs, the detachable sample pretreatment module 1 in this case is more flexible in application, thus greatly increasing its application value.

In addition, when the specimen sample is a solid sample (such as feces), the sample pretreatment module 1 of this case can provide a structural design for filtering out large particles, for example, a filter membrane is set at the bottom of the sample tank 121 to filter out the impurity particles to facilitate subsequent sample processing procedures.

On the other hand, in addition to the first connection structure exemplified in the foregoing embodiments, an adapter can also be designed or directly transmitted through a hose, so that the sample pretreatment module can be applied to various types of nucleic acid analyzers. For example, the first connection structure is essentially an output structure, which can be connected with a flexible tube (as the second connection structure) and injected into the sample port of the nucleic acid purification module, so that the sample pretreatment module and nucleic acid Fluid transfer between purification modules. In one embodiment, the output structure of the sample pretreatment module can also be set on the cover, for example, a liquid flow channel is set on the cover to introduce the sample that has been pre-extracted by the sample pretreatment module into nucleic acid purification The sample port of the module.

Fig. 7 shows a schematic diagram of a sample pre-processing module according to another embodiment of the present application, and Fig. 8 shows an exploded view of the sample pre-processing module in Fig. 7 . In another embodiment, the sample pretreatment module 7 includes a housing 71 , a tank structure 72 , a cover 73 , an output structure 74 , and a collection tube 75 . The tank structure 72 of this embodiment is the same as the tank structure 12 described above, so it will not be described again. The cover 73 is provided with a pneumatic opening 731 and an infusion pipeline 732 for driving fluid to flow between the tanks. The output structure 74 communicates with the infusion pipe 732 on the cover 73 to output the pre-treated samples. The collection tube 75 is connected to the output structure 74 to accept the samples output through the output structure 74 . For example, the collection tube 75 can be a standard product, such as a 5ml or 10ml test tube. Therefore, according to the idea of this embodiment, the sample after the initial extraction by the sample pretreatment module 7 can be directly introduced into the collection pipe 75 through the output structure 74, which can be used by the laboratory personnel to further send the sample in the collection pipe 75 to various The state-of-the-art nucleic acid analyzer performs subsequent procedures such as nucleic acid extraction, amplification, and detection.

To sum up, this case provides a detachable sample pretreatment module, which can replace the manual operation before the sample is sent to the nucleic acid analyzer, simplify the cumbersome steps required for sample pretreatment, and make sample collection, storage and transportation more convenient . In addition, the sample pretreatment module can be connected with the nucleic acid purification module to transport the pretreated samples to the nucleic acid purification module for subsequent nucleic acid extraction, amplification and detection. Furthermore, since different specimen samples require different pretreatment reagent formulations, the sample pretreatment module is designed as a detachable cartridge, and different pretreatment reagents can be configured in response to different samples. It is more flexible in application and greatly increases its application value.

Even though the present invention has been described in detail by the above-mentioned embodiments, it can be modified in various ways by those skilled in the art, all of which are within the scope of the attached patent application.

1: Sample pre-processing module 11: shell 12: Tank structure 121: sample slot 1211: Sample port 122: liquid storage tank 123: mixing tank 124: Connected pipeline 125: The first connection structure 1251: Liquid transmission pipeline 2: Nucleic acid purification module 21: Shell 22: Tank structure 23: Reaction column 24: Assign components 25: The second connection structure 3: cover body 31: Pneumatic opening 32: Infusion pipeline 33: Crossover pneumatic valve 4: Nucleic acid amplification module 41: Collecting pipe fittings 5: External cracking module 6: External active energy supply module 7: Sample pre-processing module 71: shell 72: Tank structure 73: cover body 731: Pneumatic opening 732: Infusion pipeline 74: Output structure 75: Collection pipe fittings

Figure 1 shows a schematic diagram of the assembly of the nucleic acid processing device of the present case. Figure 2 shows a schematic diagram of a nucleic acid processing device for cap-body separation. Figure 3 shows an exploded view of the nucleic acid processing device. Figures 4 and 5 show exploded views of the tank structure of the sample pretreatment module and the tank structure of the nucleic acid purification module at different angles of view. FIG. 6 shows a schematic diagram of the connection between the nucleic acid processing device and the external modules. FIG. 7 shows a schematic diagram of a sample pre-processing module according to another embodiment of the present invention. Figure 8 shows an exploded view of the sample preprocessing module in Figure 7.

12: Tank structure

121: sample slot

1211: Sample port

122: liquid storage tank

123: mixing tank

124: Connected pipeline

125: The first connection structure

1251: Liquid transmission pipeline

22: Tank structure

25: The second connection structure

Claims (14)

A nucleic acid processing device, including a sample pretreatment module and a nucleic acid purification module, wherein the sample pretreatment module includes: A sample tank has a sample inlet for adding a sample into the sample tank; A plurality of storage tanks, structured to store a plurality of pretreatment reagents, the pretreatment reagents at least include a lysate and a protease; a mixing tank, configured for mixing the sample and the pretreatment reagent; a plurality of communicating pipes configured to transport the sample and the pretreatment reagent to the mixing tank; and A first connection structure is detachably connected to a second connection structure of the nucleic acid purification module, and is configured to transport the pretreated sample to the nucleic acid purification module. The nucleic acid processing device according to claim 1, wherein the sample reservoir accommodates a sample preservation solution. The nucleic acid processing device according to claim 2, wherein the sample preservation solution is a virus transport preservation solution, a universal transport preservation solution, a bacteria transport preservation solution, or a tissue cell preservation solution. The nucleic acid processing device according to claim 1, wherein the pretreatment reagent further includes a nucleic acid precipitation aid. The nucleic acid processing device according to claim 4, wherein the nucleic acid precipitation aid is carrier RNA, ethanol, or glycogen. The nucleic acid processing device according to claim 1, wherein the first connection structure is a protrusion, and the second connection structure is a recess. The nucleic acid processing device according to claim 1 further includes a cover covering the sample pretreatment module and the nucleic acid purification module. The nucleic acid processing device according to claim 7, wherein the cover body includes a bridge pneumatic valve configured to drive the sample to flow from the sample pretreatment module to the nucleic acid purification module. The nucleic acid processing device according to claim 1, wherein the sample pretreatment module is connected to an external lysing module. The nucleic acid processing device according to claim 9, wherein the external lysis module is an ultrasonic lysis module, a heating lysis module, an optically driven lysis module, a magnetically driven particle grinding module, or a mechanical grinding module Group. The nucleic acid processing device as described in Claim 1 further includes a nucleic acid amplification module connected to the nucleic acid purification module and an external active energy supply module. The nucleic acid processing device according to claim 11, wherein the external active power supply module is an ultrasonic mixing module, a magnetically driven particle disturbance module, an optically driven heating module, or an inductive heating module. The nucleic acid processing device according to claim 1, wherein the sample is a swab, a urine, a blood, a sputum, a feces, or a saliva. A sample preprocessing module, comprising: A sample tank has a sample inlet for adding a sample into the sample tank; A plurality of storage tanks, structured to store a plurality of pretreatment reagents, the pretreatment reagents at least include a lysate and a protease; a mixing tank, configured for mixing the sample and the pretreatment reagent; a plurality of communicating pipes configured to transport the sample and the pretreatment reagent to the mixing tank; and An output structure, configured to output the pre-processed sample.

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