CN117210309A

CN117210309A - Biological sample processing device and biological detection system

Info

Publication number: CN117210309A
Application number: CN202311483504.9A
Authority: CN
Inventors: 罗严; 廖翠屏
Original assignee: Chengdu Wenjiang Maternal And Child Health Hospital; Maccura Biotechnology Co ltd
Current assignee: Maccura Biotechnology Co ltd
Priority date: 2023-11-09
Filing date: 2023-11-09
Publication date: 2023-12-12
Anticipated expiration: 2043-11-09
Also published as: CN117210309B

Abstract

The application provides a biological sample processing device and a biological detection system, and belongs to the technical field of biological sample detection. The biological sample processing device provided by the application comprises a sample processing component, a sample processing component and a biological sample processing component, wherein the sample processing component comprises a cracking cavity for receiving a sample to be processed and cracking the sample to be processed, a cleaning cavity for cleaning the sample to be processed, and a communication channel for communicating the cracking cavity and the cleaning cavity; the separation plate is arranged in the cracking cavity, the separation plate divides the cracking cavity into a first area and a second area, the bottom areas of the first area and the second area are communicated with each other, the corresponding areas of the separation plate are isolated from each other, and the first area is positioned between the communication channel and the second area. The biological sample processing device and the biological detection system at least solve the problems of high manufacturing cost and overlarge device volume of the biological sample processing device caused by the need of a large amount of lipophilic substances in the prior art.

Description

Biological sample processing device and biological detection system

Technical Field

The application belongs to the technical field of biological sample detection, and particularly relates to a biological sample processing device and a biological detection system.

Background

Nucleic acid detection has found widespread use in many fields of disease diagnosis and prognosis, public health, food safety, molecular breeding, forensic identification, and the like. Nucleic acid detection mainly comprises 3 steps: sample processing, nucleic acid extraction and purification, nucleic acid amplification and detection. The traditional laboratory nucleic acid detection requires special operators and physical partition space, and many steps require manual operation, so that the problems of complex operation exist, and the risks of easy harm to operators caused by samples and reagents and pollution to nucleic acid products are also faced.

The fully-integrated automatic nucleic acid detection system can well solve the problems, realizes the detection target of 'sample in-out', greatly reduces personnel operation, reduces potential pollution hazard of samples and amplified products, improves the detection reliability, saves labor cost to the greatest extent and reduces space occupation.

The portable full-integrated nucleic acid detection, also called "point-of-care testing", is a portable, miniaturized and automatic nucleic acid detection system, and uses external automation equipment to simulate manual operation so as to implement full-automatic process from sample treatment to nucleic acid detection.

Conventional molecular diagnostic POCTs require at least an integrated sample processing chamber, an extraction purification chamber, and an amplification detection chamber, which are disposed at intervals. The sample treatment cavity is mainly used for carrying out the lysis treatment on the sample, namely releasing DNA in the sample such as cells, tissues, blood and the like, and sometimes removing inhibitors so as to carry out the subsequent PCR analysis; the extraction and purification cavity is used for extracting and purifying nucleic acid, plays a key role in the PCR process, and can effectively separate target nucleic acid, remove pollutants, improve the efficiency of the PCR reaction and reduce the background interference, thereby ensuring the accuracy, the sensitivity and the specificity of the PCR reaction; and the amplification detection cavity is used for amplifying the purified nucleic acid and detecting the result.

In order to realize full automation of a detection process in the prior art, a molecular diagnosis POCT device needs to move a substance to be detected between each cavity to realize operation steps of different stages, usually adopts adsorption of the substance to be detected by magnetic particles, and then moves the magnetic particles which have captured the substance to be detected in liquid in each cavity by external magnetic field movement, but the mode is only in a theoretical stage, and an actual product still needs to consider the sealing problem between each cavity in the transportation process and how to avoid the problem of no loss of the magnetic particles in the moving process between each cavity.

Disclosure of Invention

During transportation of the molecular diagnostic POCT consumable cartridge, and before the cartridge is put into the instrument for operation, the liquid needs to be prevented from flowing mutually between the chambers, for example, the pyrolysis liquid in the pyrolysis chamber cannot enter the washing chamber at the rear end, and the washing liquid in the washing chamber cannot flow into the pyrolysis chamber. Therefore, a barrier device capable of blocking the movement of liquid is required between different sample processing chambers. Filling paraffin is adopted as a valve, so that on one hand, the paraffin can be solidified to form a blocking effect so as to be convenient for storage and transportation; on the other hand, paraffin is a lipophilic substance floating on the surface of the liquid and does not participate in the reaction of nucleic acid detection, and meanwhile, the paraffin also has the function of reducing the friction force between the magnetic particles and the supporting surface in the moving process.

The paraffin can be liquefied at a lower temperature to cover the liquid surface of each chamber, the external magnetic field enables the magnetic particles to pass through the liquid paraffin first in the moving process of each chamber, and the liquid paraffin layer needs to be thick enough to realize that the paraffin wraps the magnetic particles in the process of passing the magnetic particles, so that the magnetic particles play a role in lubrication. For example, the magnetic particles used as the extraction and purification reagent have a lubricating effect due to the lipophilic property of the liquefied paraffin during the transfer process, so that the phenomenon that the magnetic particles are removed during the transfer process, namely the problem of friction force or roughness between the magnetic particles and the inner surface of the consumable cartridge during the movement process, is reduced during the movement process of the magnetic particles, and the amount of the substance to be detected finally moved into the detection cavity is reduced, so that the accuracy of the detection result is greatly reduced.

The embodiment of the application provides a biological sample processing device and a biological detection system, which can reduce friction force in a moving process by utilizing the lipophilicity of paraffin under the action of solid paraffin obstruction. And in the process of using the paraffin, the use amount of the paraffin can be reduced, so that the time for melting the solid paraffin is reduced, the detection efficiency is improved, the manufacturing cost of the biological sample processing device is reduced, and the device is miniaturized.

A first aspect of an embodiment of the present application provides a biological sample processing device, comprising: a sample processing member; the device comprises a cracking cavity for receiving a sample to be treated and cracking the sample to be treated; a cleaning chamber for cleaning a sample to be processed; and a communication channel communicating the lysis chamber and the wash chamber; the lysis cavity is used for containing an aqueous solution containing a sample to be treated; the separation plate is arranged in the cracking cavity, the separation plate divides the cracking cavity into a first area and a second area, the bottom areas of the first area are communicated with each other, the corresponding areas of the separation plate are isolated from each other, and the first area is positioned between the communication channel and the second area; when the lysis cavity contains an aqueous solution containing a sample to be treated, one end of the baffle plate stretches into the lower part of the liquid surface of the aqueous solution; a lipophilic substance which can change shape along with the change of temperature is also arranged between the first area and the tail end of the communication channel; the lipophilic substance isolates the liquid flow of the cracking cavity and the cleaning cavity in a solid state at normal temperature; the lipophilic material is overlaid on top of the aqueous solution in liquid form, the lipophilic material being immiscible with the aqueous solution.

Further, the biological sample processing device further comprises a magnetic conduction piece, wherein the magnetic conduction piece is accommodated in at least one of the cracking cavity, the cleaning cavity and the communication channel, and the magnetic conduction piece is used for entering the cracking cavity under the action of an external magnetic field and adsorbing target substances in a sample to be processed.

Further, the lipophilic substance is solid at a first temperature and is at least partially accommodated in the communication channel, and the magnetic conductive piece is fixedly sealed in the lipophilic substance.

Further, the lipophilic substance may be melted at a second temperature so as to flow into the first region and float above the sample to be treated accommodated in the lysis chamber, the second temperature being greater than the first temperature.

Further, the magnetic conduction piece positioned in the communication channel can penetrate through the lipophilic substance to enter the cracking cavity and contact with the sample to be processed to capture the target substance under the action of an external magnetic field, and penetrate through the lipophilic substance to the cleaning cavity.

Further, the communication channel includes a first port adjacent the lysing chamber and a second port adjacent the purging chamber;

the magnetic conduction piece is fixedly sealed at the first port or the second port through the lipophilic substance.

Further, the cracking cavity comprises a cavity bottom wall and a plurality of cavity side walls formed by bending and extending the cavity bottom wall, the cavity side walls comprise a first area side wall and a second area side wall which are arranged at intervals opposite to the partition board, the first area side wall is adjacent to the communication channel, and the interval between the partition board and the first area side wall is smaller than the interval between the partition board and the second area side wall.

Further, the communicating channel, the cracking cavity and the cleaning cavity are provided with openings in the same direction, and the sample adding component further comprises a cover plate covering the openings;

the cover plate and the partition plate are arranged at intervals;

or, the cover plate is abutted with the partition plate.

Further, the biological sample processing device further comprises a sample adding member at least partially inserted into the second region, wherein the sample adding member is provided with a containing cavity for containing a sample to be processed and a filtering outlet communicated with the containing cavity and the second region.

Further, the sample processing member further comprises an opening communicating with the communication channel, the lysis chamber and the washing chamber, the sample adding member further comprises a cover plate covering the opening, and the partition plate is connected with the cover plate.

Further, the cracking cavity comprises a cavity bottom wall and a plurality of cavity side walls formed by bending and extending from the cavity bottom wall, and the partition plate is in interference fit with the cavity side walls in the width direction of the partition plate.

Further, the cracking cavity comprises a cavity bottom wall, a plurality of cavity side walls formed by bending and extending from the cavity bottom wall, and a guide rail formed by extending inwards from the cavity side walls, and the partition board is clamped on the guide rail.

Further, a cracking assisting piece for cracking the sample to be processed is prestored in the cracking cavity.

Further, a preset distance is reserved between the bottom of the partition plate and the bottom of the cracking cavity, so that the bottom areas of the first area and the second area are communicated with each other.

Further, a biological detection system comprising: a biological sample processing device; and

The magnet is abutted against the side wall of the outer chamber of the sample processing component and is used for providing an external magnetic field; and/or

The heating device is abutted against the side wall of the outer cavity of the cleaning cavity and/or the communication channel and is used for heating; and/or an ultrasonic device which is abutted to the side wall of the outer chamber of the cracking chamber and is used for transmitting ultrasonic waves to the cracking chamber and/or the communication channel.

The technical scheme provided by the embodiment of the application at least has the following beneficial effects:

the application provides a biological sample processing device, in the biological sample processing device, a partition board is arranged in a cracking cavity, the cracking cavity is divided into a first area and a second area which are mutually communicated in a bottom area and mutually isolated in a corresponding area of the partition board by utilizing the partition board, so that paraffin with an isolation function does not need to cover the liquid surface of the whole cracking cavity, but only needs to cover the liquid surface of the first area, thus, the required paraffin layer thickness is ensured, the paraffin consumption is reasonably reduced, the volume of a card box is reduced, and the miniaturization of products is realized. Meanwhile, the dosage of the solid paraffin in the blocking process is reduced, and the time for forming the liquid paraffin by melting the paraffin at high temperature in the detection process is shortened, so that the pretreatment time is shortened, and the detection speed of the biological sample processing device is improved.

The present application also provides a biological detection system which can also reduce manufacturing costs and miniaturize the device since the biological detection system includes the biological sample processing device of the foregoing technical scheme.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the application as claimed.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are needed in the embodiments of the present application will be briefly described below, and it is obvious that the drawings described below are only some embodiments of the present application, and other drawings may be obtained according to these drawings without inventive effort to a person of ordinary skill in the art.

FIG. 1 is a schematic perspective view of a biological sample processing device according to some embodiments of the present application;

FIG. 2 is a cross-sectional view of the biological sample processing device of FIG. 1 taken along line A-A;

FIG. 3 is an enlarged view of portion B of the biological sample processing device of FIG. 2;

FIG. 4 is a schematic perspective view of a sample processing member in a biological sample processing device according to some embodiments of the present application;

FIG. 5 is a schematic perspective view of a sample application member of a biological sample processing device according to some embodiments of the present application;

FIG. 6 is a schematic representation of a prior art lipophilic material coverage;

fig. 7 is a schematic view of coverage of lipophilic substances in the present embodiment.

Wherein, the reference numerals are as follows:

100. a biological sample processing device; 10. a sample processing member; 102. a first region; 103. a second region; 11. a lysing chamber; 111. a chamber bottom wall; 112. a chamber sidewall; 1121. a first region sidewall; 1122. a second region sidewall; 12. cleaning the cavity; 13. a communication passage; 131. a first port; 132. a second port; 14. an opening; 20. a partition plate; 30. A sample addition member; 31. a housing chamber; 32. a filter outlet; 33. a cover plate; 40. a lipophilic substance; 50. a magnetic conductive member; 300. Paraffin wax; 400. the sample to be treated.

Detailed Description

Features and exemplary embodiments of various aspects of the application are described in detail below. In the following detailed description, numerous specific details are set forth in order to provide a thorough understanding of the application. It will be apparent, however, to one skilled in the art that embodiments of the application may be practiced without some of these specific details. The following description of the embodiments is merely intended to provide a better understanding of the application by showing examples of the application.

In the description of the present application, it should be understood that the direction or positional relationship indicated with respect to the description of the orientation, such as up, down, back, etc., is based on the direction or positional relationship shown in the drawings, and is merely for convenience in describing the embodiments of the present application and simplifying the description, and does not indicate or imply that the apparatus or element referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the embodiments of the present application.

In the description of the embodiments of the present application, plural means two or more, and greater than, less than, etc. are understood to exclude this number. The description of the first and second is for the purpose of distinguishing between technical features only and should not be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated or implicitly indicating the precedence of the technical features indicated.

In the description of the embodiments of the present application, unless explicitly defined otherwise, terms such as arrangement, installation, connection, etc. should be construed broadly, and those skilled in the art may reasonably ascertain the specific meaning of the terms in the present application by combining the specific contents of the technical solutions.

It should be noted that, without conflict, the embodiments of the present application and features of the embodiments may be combined with each other. The embodiments will be described in detail below with reference to the accompanying drawings.

Molecular diagnosis POCT is a detection mode which is carried out on a sampling site and can rapidly obtain a detection result by using a portable analysis instrument and a matched reagent, and the main standard is a detection site which does not need to be fixed, and the reagent and the instrument are portable and can be operated in time.

In this regard, a biological sample processing device is usually required at the detection site to perform operations such as lysis, washing, amplification, etc. on the collected biological sample, so that the portable analysis apparatus at the site can accurately identify the information of the collected biological sample, and thus make a corresponding judgment. However, in the prior art, the lysis step and the washing and amplification steps are usually performed in different instruments, so that the immediate detection effect of sample inlet and sample outlet cannot be really realized.

Molecular diagnostic POCT is performed by integrating lysis, washing, amplification and detection into different chambers of the same consumable cartridge, and typically connecting paraffin in different sample processing chambers to each other to form a wax channel. The design of such wax channels often requires a large amount of paraffin in order to achieve sealing during solidification, communication during liquefaction and lubrication. While a large amount of paraffin has some problems: since a large amount of paraffin is in a solid state at the time of transportation or storage, it is necessary to liquefy paraffin in a solid state during the detection, and a large amount of paraffin takes more time to change from solid to liquid, which is contrary to the rapid detection pursued by molecular diagnostic POCT. Therefore, how to provide a biological sample processing device, reduce the paraffin consumption, and simultaneously realize miniaturization of products as much as possible is a problem to be solved by those skilled in the art. Because the biological sample processing device at least needs to perform some of the above steps, the biological sample processing device generally includes a plurality of sample processing chambers disposed at intervals and a communication channel for communicating with adjacent sample processing chambers, and meanwhile, a lipophilic substance is usually placed in the communication channel and/or the sample processing chamber to isolate different detection solutions (the detection solutions may be samples to be processed, cleaning solutions, amplification reagents, etc.) in different sample processing chambers.

If the lysis step is to be built in the biological sample processing device, a sample to be processed with a larger volume (the sample to be processed is mixed with a sample collected from an organism) needs to be added into the sample processing cavity, so that the volume of the sample processing cavity is larger, the height is limited, and the cross-sectional area of the sample processing cavity is larger, so that the requirement for lipophilic substances covering the sample processing cavity is large, the volume of the biological sample processing device is further increased, and the miniaturization is not facilitated.

As described above, since the capacity of each sample processing chamber is small, in some larger sample processing chambers, it is generally necessary to cover the entire surface of the sample processing chamber with a lipophilic substance, so that on one hand, the amount of the lipophilic substance is increased, resulting in increased cost; on the other hand, due to the increase of lipophilic substances, it is accordingly necessary to increase the size of the communication channel for accommodating the lipophilic substances.

Based on the above-mentioned considerations, in order to avoid the increase of the amount of lipophilic substances caused by the excessive volume of the individual sample processing chambers, the inventors have conducted intensive studies to design a sample processing apparatus, and by disposing a partition plate in the cleavage chamber, the lipophilic substances only need to cover the surface of the first region, so as to solve the problems of excessive amount of lipophilic substances and excessive volume of the sample processing apparatus.

The biological sample processing device and the biological detection system provided by the application can be applied to clinical detection in hospitals, rapid detection in private clinics, household self-detection, detection outside clinical laboratory, scattered detection and the like.

The technical scheme of the biological sample processing device and the biological detection system provided by the specific embodiment of the application is further described below.

FIG. 1 is a schematic perspective view of a biological sample processing device according to some embodiments of the present application; FIG. 2 is a cross-sectional view of the biological sample processing device of FIG. 1 taken along line A-A; FIG. 3 is an enlarged view of portion B of the biological sample processing device of FIG. 2; FIG. 4 is a schematic perspective view of a sample processing member in a biological sample processing device according to some embodiments of the present application; fig. 5 is a schematic perspective view of a sample application member of a biological sample processing device according to some embodiments of the present application.

Referring to fig. 1 to 5, in a first aspect, a biological sample processing device 100, i.e., a consumable cartridge, is provided according to an embodiment of the present application, which includes a sample processing member 10 and a partition 20. Wherein the sample processing member 10 includes a lysis chamber 11 for receiving and lysing the sample to be processed, a washing chamber 12 for washing the sample to be processed, and a communication passage 13 communicating the lysis chamber 11 and the washing chamber 12; the partition board 20 is disposed in the cracking chamber 11, the partition board 20 divides the cracking chamber 11 into a first area 102 and a second area 103, the bottom areas of the first area are communicated with each other, the corresponding areas of the partition board 20 are isolated from each other, and the first area 102 is located between the communication channel 13 and the second area 103.

In some embodiments of the present application, the biological sample processing device 100 may be a consumable and used to detect biological samples in cooperation with other functional instruments, analytical instruments; in other embodiments of the present application, the biological sample processing device 100 may be a reusable product, and after a single use, the biological sample processing device 100 may be reused by washing, sterilizing, refilling with a test fluid, and packaging.

The biological sample processing device 100 may be made of transparent materials such as plastic and glass, so as to facilitate the operation and observation of medical staff.

The sample processing member 10 includes at least a lysis chamber, a washing chamber, and an amplification chamber.

Wherein the cracking cavity is used for cracking the sample to be treated entering the cracking cavity so as to release target substances in the sample to be treated. In an embodiment, the target substance is a nucleic acid and the lysis chamber functions to destroy the sample and release the nucleic acid in the sample into the lysis chamber.

The release of the target substance may be performed by physical release, for example, in some embodiments of the present application, a certain amount of the lysis particles may be packaged in advance in the lysis chamber, and after the sample to be processed enters the lysis chamber, the functional instrument may be used to drive the lysis particles to move in the lysis chamber, grind the sample and release the target substance.

In these embodiments of the application, the cracking particles may be polystyrene particles, borosilicate glass particles, quartz glass particles, or the like; the functional instrument can select an ultrasonic device, and the ultrasonic device is utilized to release ultrasonic waves, so that the cracking particles can generate rapid vibration so as to grind the sample to be treated and better release target substances.

The cleaning cavity is internally provided with cleaning liquid and is used for cleaning other substances except target substances, such as inhibitor and lysate in the lysis cavity, so as to improve the detection accuracy.

The amplification chamber contains lyophilized reaction components and a reconstituted solution of a bioassay and is used to amplify the target substance in an increased amount for monitoring analysis.

The lysis chamber 11 and the wash chamber 12 may be any two adjacent chambers among the lysis chamber, the wash chamber, the amplification chamber, and other chambers not mentioned. Meanwhile, the size, the side wall shape and the bottom wall shape of each sample processing cavity are not limited, and can be selected according to actual conditions.

For example, the volume of each sample processing chamber may be determined according to the amount of the detection liquid to be stored or whether other structures are required to be added into the chamber; in order to facilitate the transportation of substances in the sample processing cavity, a rounding angle between the side wall and the bottom wall can be arranged; to facilitate placement of biological sample processing device 100, multiple sample processing chambers may be provided with bottom walls that are in the same plane or in different planes, and so forth.

The lysis chamber 11 is adapted to receive a sample to be processed, and in a possible embodiment, the lysis chamber 11 may be adapted to receive an externally entered sample to be processed, and may also be adapted to receive a sample to be processed that has been processed by another sample processing chamber.

The partition 20 is disposed in the cracking chamber 11, and in a possible embodiment, the partition 20 may be directly formed in the cracking chamber 11 to directly partition the cracking chamber 11 into a first area 102 and a second area 103 with bottom areas mutually communicated and corresponding areas of the partition 20 mutually isolated;

or the first area 102 and the second area 103 which are detachably connected with the cracking chamber 11 and are tightly matched with the cracking chamber 11 and are used for separating the cracking chamber 11 into the first area 102 and the second area 103 which are mutually communicated with the bottom area and mutually isolated from the corresponding areas of the partition board 20 can be selected according to the actual conditions of production.

The first region 102 communicates with the bottom region of the second region 103, possibly in an embodiment where at least part of the bottom of the partition 20 is structured such that the first region 102 communicates with the second region 103. For example, the bottom end of the partition 20 may be provided with a through hole communicating the first region 102 with the second region 103, the first region 102 communicating with the second region 103 through the through hole, and capable of mass exchange.

In these embodiments of the present application, the first region 102 and the bottom region of the second region 103 are communicated with each other, so that the detection liquid in the lysis chamber 11 communicates between the first region 102 and the second region 103 to ensure the function of the lysis chamber 11.

The first region 102 is located between the communication channel 13 and the second region 103, so that the sealing area of the cracking chamber 11 can be effectively reduced.

Illustratively, taking the lysing chamber 11 as a lysing chamber and the cleaning chamber 12 as a cleaning chamber, the baffle 20 divides the lysing chamber into a first region 102 and a second region 103.

In order to prevent the detection liquid pre-stored in the cracking cavity from flowing into the cleaning cavity through the communication channel 13, a layer of lipophilic substance needs to be paved on the upper surface of the cracking cavity so as to prevent the detection liquid in the cracking cavity from entering the cleaning cavity through the communication channel 13, and thus, the partition board 20 is arranged in the cracking cavity and divides the cracking cavity into the first area 102 and the second area 103, and the lipophilic substance only needs to be paved on the upper surface of the first area 102 communicated with the communication channel 13, so that the use of the lipophilic substance is greatly reduced, and the manufacturing cost is reduced. Meanwhile, the use amount of the lipophilic substance is reduced, so that the space of the communication channel 13 can be reduced, and further miniaturization is realized.

The movement of the sample to be treated in the lysis chamber 11, the washing chamber 12 and the communication channel 13 is realized by moving other members after the sample to be treated is adsorbed by other members with the help of other members. Thus, the lipophilic substance on the one hand has the function of forming a liquid environment in the communication channel 13 and each sample processing cavity, so that other components can smoothly move in each sample processing cavity or the communication channel 13 without being stuck to the inner wall of the sample processing cavity or the communication channel 13 under the action of friction force; the other aspect is to separate the detection liquid in each sample processing chamber.

That is, the arrangement of the lipophilic substance can optimize the transfer efficiency of the sample to be processed and simultaneously avoid the circulation of the detection liquid of each sample processing cavity.

In some embodiments of the present application, if desired, a partition 20 may be provided in the lysis chamber 11, the wash chamber 12, and other sample processing chambers simultaneously to further reduce the amount of lipophilic substances used, thereby miniaturizing the various sample processing chambers and miniaturizing the biological sample processing device 100.

In the biological sample processing device 100 provided in the above embodiment of the present application, by disposing the partition board 20 in the lysis chamber 11 and dividing the lysis chamber 11 into the first region 102 and the second region 103 with the bottom regions thereof being mutually communicated and the corresponding regions of the partition board 20 being mutually isolated by the partition board 20, the lipophilic substance having the isolation function does not need to cover the entire surface of the lysis chamber 11, but only needs to cover the surface of the first region 102, thereby greatly reducing the use amount of the lipophilic substance and reducing the manufacturing cost. Meanwhile, the use amount of the lipophilic substance is reduced, so that the space of the communication channel 13 can be reduced, and further miniaturization is realized.

In some embodiments of the present application, the biological sample processing device 100 further comprises a loading member 30, wherein the loading member 30 is at least partially inserted into the second region 103, and the loading member 30 has a receiving cavity 31 for receiving a sample to be processed and a filtering outlet 32 communicating the receiving cavity 31 with the second region 103.

The loading member 30 is used for loading the sample to be processed. In some embodiments of the application, the loading member 30 may be used to load the sample to be processed collected by the external member into the second region 103 of the lysis chamber; the sample to be treated which is subjected to the cracking treatment by the external component can also be added into the cleaning cavity; it can also be used to add a sample to be treated, which has been subjected to a washing treatment by an external member, to an amplification chamber.

The loading member 30 is at least partially inserted into the second region 103, and it may be an embodiment that the loading member 30 is at least partially disposed in the second region 103, where the loading member 30 may be detachably connected, fixedly connected, or even integrally formed with the lysis chamber 11, and may be selected according to practical situations.

The accommodating cavity 31 is used for accommodating a sample to be processed, and the filtering outlet 32 is communicated with the accommodating cavity 31 and the second area 103. Illustratively, in embodiments where the loading member 30 is fixedly coupled to the lysis chamber 11, the receiving chamber 31 may include a loading port (not shown) through which the sample to be processed may be loaded into the receiving chamber 31; in the embodiment in which the sample loading member 30 is detachably connected to the lysis chamber 11, besides the foregoing arrangement, the filter outlet 32 may be detachably provided, and in the sample loading stage, the receiving chamber 31 is exposed by removing the filter outlet 32, and after the sample to be processed is loaded, the filter outlet 32 is installed.

The sample application member 30 may be configured to filter the sample to be processed before it enters the lysis chamber 11 to remove unwanted material, and the lysis chamber 11 may be designed to be larger to accommodate the sample application member 30. Thus, in these embodiments of the present application, the amount of lipophilic material used is reduced by providing the partition 20 to the cleavage chamber 11 such that the cleavage chamber 11 is partitioned by the partition 20.

In some embodiments of the present application, the biological sample processing device 100 further comprises a lipophilic substance 40 for forming a barrier between the lysis chamber 11 and the wash chamber 12, the lipophilic substance 40 being mutually insoluble with the sample to be processed contained in the lysis chamber 11.

The lipophilic substance 40 is not compatible with the sample to be treated contained in the lysis chamber 11, and in a possible embodiment, the lipophilic substance 40 does not react with the sample to be treated and the detection liquid contained in the lysis chamber 11, and the lipophilic substance 40 is layered with the detection liquid, i.e., the lipophilic substance 40 floats above the liquid surface of the detection liquid, and does not form an emulsion under natural conditions. Similarly, the lipophilic material 40 is also insoluble in the liquid contained in the wash chamber 12. The lipophilic substance 40 serves to separate the liquid, such as a washing liquid, in the lysis chamber 11 from the washing chamber 12, and by the separation of the lipophilic substance 40, a barrier is formed between the lysis chamber 11 and the washing chamber 12, preventing free flow of liquid between a lysis chamber 11 and the washing chamber 12, and mixing. The lipophilic substance 40 also prevents the detection liquid in the lysis chamber 11 and the wash chamber 12 from volatilizing.

By way of example, the lipophilic substance 40 may be a paraffin, oil, or other lipophilic material. In some embodiments of the present application, in order to enable the lipophilic material 40 to maintain a certain shape when the biological sample processing device 100 is not in use, so as to avoid displacement during transportation, paraffin may be selected as the lipophilic material 40, and the paraffin is melted by heating when it is in use due to its solid state at normal temperature. The following description will take the lipophilic substance 40 as paraffin as an example.

In some embodiments of the present application, the biological sample processing device 100 further includes a magnetic conductive member 50, where the magnetic conductive member 50 is accommodated in at least one of the lysis chamber 11, the washing chamber 12 and the communication channel 13, and the magnetic conductive member 50 is used to enter the lysis chamber 11 and adsorb target substances in the sample to be processed under the action of an external magnetic field.

The magnetic conductive member 50 is used for adsorbing a target substance and carrying the target substance by an external magnetic field, i.e., transferring the target substance between different sample processing chambers through the lipophilic substance 40, so as to carry the adsorbed target substance to the different sample processing chambers for different processes. The number of the magnetic conductive members 50 may be plural, and the target substance is adsorbed by the plurality of magnetic conductive members 50 at the same time, so as to improve the adsorption and transfer efficiency of the target substance.

Illustratively, in some embodiments of the present application, the magnetically permeable member 50 may be superparamagnetic magnetic particles.

In some embodiments of the present application, the external magnetic field may be a permanent magnet or an electromagnet, and the plurality of magnetic conductive members 50 are attracted by the external magnetic field, and the external magnetic field is moved along the structure of the sample processing member 10 to move the plurality of magnetic conductive members 50 into the sample to be processed in the lysis chamber 11, and after the plurality of magnetic conductive members 50 attract the target substances, the external magnetic field is moved along the sample processing member 10, so that the target substances sequentially pass through the plurality of sample processing chambers.

In some embodiments of the present application, the lipophilic material 40 is solid at the first temperature and is at least partially contained within the communication channel 13, and the magnetic permeable member 50 is secured within the communication channel 13 by the lipophilic material 40.

The lipophilic substance 40 is solid at the first temperature, and in a possible embodiment, the environmental temperature is maintained at about the first temperature during storage and transportation of the biological sample processing device 100 after production is completed, and the lipophilic substance 40 is in a solid state. In other words, the lipophilic substance 40 may be provided to remain in a solid state at normal temperature. Wherein the specific value of the first temperature is related to the type of the lipophilic substance 40, and thus the type of the lipophilic substance 40 can be determined according to the use environment of the biological sample processing apparatus 100.

Illustratively, the melting point of the lipophilic material 40 may be selected to be 20 ℃ to 40 ℃ above the first temperature to avoid melting of the lipophilic material 40 at normal temperatures (10 ℃ to 40 ℃).

Because the magnetic conductive member 50 is used for adsorbing the target substance and moves under the action of the external magnetic field, the magnetic conductive member 50 cannot enter the sample processing cavity to contact with the detection liquid prematurely, so that the influence on the magnetic conductive member 50 or the sample to be processed is avoided. For example, if the magnetic conductive member 50 enters the cracking chamber and contacts with the detection liquid prematurely, during the cracking process, the magnetic conductive member 50 is easy to collide and rub with the cracking particles under the action of the functional instrument, so as to damage the surface of the magnetic conductive member 50 and affect the adsorption effect of the subsequent target substances.

Based on this, in some embodiments of the present application, a magnetic conductive member 50 may be provided to be fixed in the communication channel 13 by the lipophilic substance 40. In this manner, magnetic flux conducting member 50 may be secured when biological sample processing device 100 is not in use, preventing magnetic flux conducting member 50 from moving within sample processing member 10 before biological sample processing device 100 is used.

In some embodiments of the present application, the lipophilic substance 40 may be melted at a second temperature to flow into the first region 102 and float above the sample to be treated contained in the lysis chamber 11, the second temperature being greater than the first temperature.

The second temperature is higher than the melting point of the lipophilic substance 40, and during the use of the biological sample processing apparatus 100, an external functional device (such as a heating device) may be used to heat the lipophilic substance 40, so that the lipophilic substance 40 melts and flows into the lysis chamber 11, and at this time, the lipophilic substance 40 in a liquid state floats on the detection liquid (to-be-processed sample) in the lysis chamber 11 (because the paraffin density is lower than the detection liquid and is insoluble with the detection liquid, the paraffin can float above the detection liquid) and is located above the to-be-processed sample, thereby isolating the detection liquid in the lysis chamber 11 from the detection liquid in the cleaning chamber 12.

In some embodiments of the present application, the magnetic conductive member 50 located in the communication channel 13 may pass through the lipophilic substance 40 into the lysis chamber 11 and contact the sample to be processed under the action of an external magnetic field.

The lipophilic substance 40 enters the lysis chamber 11 and contacts the sample to be treated, possibly by moving the magnetic conductive element 50 with an external magnetic field after the completion of the functioning of the functional instrument (or just before the completion) such as the heating means, the ultrasound means, etc.

Illustratively, taking the cleavage cavity 11 as an example of the cleavage cavity, after the heating device heats and melts the lipophilic substance 40 and the ultrasonic device processes the sample to be processed in the cleavage cavity, the magnetic conductive member 50 located in the communication channel 13 may be moved into the cleavage cavity by an external magnetic field to adsorb the released target substance.

In some embodiments of the application, the communication channel 13 comprises a first port 131 adjacent to the lysis chamber 11 and a second port 132 adjacent to the wash chamber 12; the magnetic conductive member 50 is fixed to the first port 131 or the second port 132 by the lipophilic material 40.

The first port 131 is adjacent to the cracking chamber 11, and it is possible to implement that the first port 131 is located at one end of the communication channel 13 near the cracking chamber 11; the second port 132 is adjacent to the wash chamber 12, and it is possible that the second port 132 is located at an end of the communication channel 13 near the wash chamber 12. The plane in which the first port 131 and the plane in which the second port 132 are located may be parallel to each other or may be in an intersecting relationship.

For example, the communication channel 13 may be a straight tube, where the first port 131 and the second port 132 are located at two ends of the communication channel 13, and a plane where the first port 131 and the second port 132 are located are parallel to each other.

The magnetic conductive member 50 is fixedly sealed to the first port 131 or the second port 132 by the lipophilic substance 40, and in a possible implementation manner, the magnetic conductive member 50 is disposed near the cracking chamber 11 or the cleaning chamber 12 in the communication channel 13, so that the lipophilic substance 40 quickly enters the cracking chamber 11 or the cleaning chamber 12 after being melted and floats above the detection liquid to further play a role in separation, and meanwhile, the magnetic conductive member 50 is positioned at one end of the communication channel 13 and can more quickly enter the cracking chamber 11 or the cleaning chamber 12.

In some embodiments of the present application, the magnetically permeable member 50 comprises magnetic particles (magnetic particles) or magnetically permeable pellets or the like. Magnetic particles refer to very small particles or aggregates of particles that have magnetic properties. They are generally composed of magnetic materials such as iron, nickel, cobalt, etc. The size of the magnetic particles is typically on the order of nanometers to micrometers. The magnetic particles have special magnetic behaviors and can show magnetization behaviors under the action of an externally applied magnetic field. Specifically, when magnetic particles are exposed to an external magnetic field, the magnetic moment inside the particles is affected by the magnetic field, resulting in the particles themselves also exhibiting magnetism. Such magnetization behavior may be permanent (hard magnetic) or reversible (soft magnetic), depending on the properties of the magnetic material. In this embodiment, the magnetic particles are mainly used in biomedical separations and the like.

It is noted that the application and research of magnetic particles also requires consideration of their impact on the environment and health, especially at the nanoscale. Thus, related safety measures and regulations need to be complied with when using and handling magnetic particles.

In some embodiments of the present application, the cracking chamber 11 includes a chamber bottom wall 111, and a plurality of chamber side walls 112 formed by bending and extending from the chamber bottom wall 111, and the chamber side walls 112 include a first region side wall 1121 and a second region side wall 1122 disposed opposite to the partition 20, the first region side wall 1121 is adjacent to the communication channel 13, and a space between the partition 20 and the first region side wall 1121 is smaller than a space between the partition 20 and the second region side wall 1122.

The first area sidewall 1121 is adjacent to the communication channel 13, and it may be implemented that the first area sidewall 1121 is the sidewall closest to the communication channel 13, and the second area sidewall 1122 is the sidewall farthest from the communication channel 13 because the second area sidewall 1122 is disposed opposite to the first area sidewall 1121.

Based on this, the distance between the partition 20 and the first region side wall 1121 is smaller than the distance between the partition 20 and the second region side wall 1122, and it is possible that the partition is disposed closer to the communication passage 13. Thus, the volume of the first region 102 is smaller than that of the second region 103, so that the second region 103 has a larger accommodating space while the consumption of the lipophilic substance 40 is reduced.

In some embodiments of the present application, the sample processing member 10 further comprises an opening 14 communicating with the channel 13 and the lysis chamber 11 and the wash chamber 12, and the sample application member 30 further comprises a cover plate 33 covering the opening 14.

It should be noted that, the above description of the present embodiment is based on the description that the sample processing member 10 includes only the lysis chamber 11 and the washing chamber 12, and in the case that the sample processing member 10 further includes the third sample processing chamber and the fourth sample processing chamber, the opening 14 can also communicate with the third sample processing chamber and the fourth sample processing chamber.

Based on this, the cover plate 33 covers the opening 14 by welding so that the biological sample processing device 100 forms a sealed space. Before the cover plate 33 does not cover the opening 14, the sample processing chambers and the communication channels 13 can be cleaned and disinfected through the opening 14, and the detection liquid is pre-stored in each sample processing chamber, the magnetic conductive piece 50 is pre-stored in the communication channel 13, and the solid lipophilic substance 40 is pre-stored in each sample processing chamber and/or the communication channel 13.

After the cover plate 33 covers the opening 14, the cover plate 33 and the partition plate 20 can be arranged at intervals, the cover plate 33 and the partition plate 20 can also be abutted, and the partition plate 20 is vertically connected to the cover plate 33, so that the cover plate 33 can be welded, clamped or integrally formed, and the selection is in consideration of the processing technology and the cost, and does not influence the practical innovation point of the application. In these embodiments of the application, the abutment between the cover plate 33 and the separator 20 may be selected for ease of tooling. By providing an abutment between the cover plate 33 and the partition plate 20, the design of the mold can be simplified, and molding can be facilitated.

Meanwhile, in the embodiment where the cover plate 33 is spaced from the partition 20, that is, the cover plate 33 is not connected to the partition 20, there is a portion of the lipophilic substance 40 entering the second region 103 through the gap between the cover plate 33 and the partition 20, and the lipophilic substance 40 floats above the detection liquid in both the first region 102 and the second region 103, so that no influence is generated.

In some embodiments of the present application, the sample processing member 10 further comprises an opening 14 communicating with the channel 13 and the lysis chamber 11 and the wash chamber 12, and the sample application member 30 further comprises a cover plate 33 covering the opening 14; the partition 20 is connected to the cover 33.

Based on this, the partition board 20 is directly connected with the cover board 33, so that the molding difficulty of the partition board 20 is reduced, and meanwhile, when the cover board 33 does not cover the opening 14, the addition of detection liquid into each sample processing cavity, the addition of cracking particles into the cracking cavity, the addition of freeze-dried reaction components into the amplification cavity and the like are facilitated.

The connection between the partition 20 and the cover 33 may be adhesive, riveted, integrally formed, or the like.

In some embodiments of the present application, the cleavage chamber 11 includes a chamber bottom wall 111 and a plurality of chamber side walls 112 formed by bending and extending from the chamber bottom wall 111, and the partition 20 is in interference fit with the chamber side walls 112 in the width direction thereof.

Based on this, in some embodiments of the present application, the partition 20 may be provided with a width slightly larger than the space between the chamber side walls 112 on both sides in the width direction thereof, so that both sides in the width direction of the partition 20 are closely fitted to the chamber side walls 112 after the cover plate 33 covers the opening 14, to avoid a situation that the lipophilic substance 40 passes through the gap between the partition 20 and the chamber side walls 112 from the first region into the second region during the subsequent use.

In some embodiments of the present application, the lysis chamber 11 includes a chamber bottom wall 111, a plurality of chamber side walls 112 formed by bending and extending from the chamber bottom wall 111, and a rail (not shown) formed by extending inward from the chamber side walls 112, and the partition 20 is clamped on the rail.

By providing rails on the chamber side walls 112, the use of the rails to provide guidance for the insertion of the partition 20 facilitates subsequent covering of the opening 14 with the cover 33.

In some embodiments of the application, the number of rails may be one, with one rail being provided on one side of the sample processing chamber on chamber side wall 112, and with a single-sided track providing guidance for partition 20 during subsequent covering of opening 14 by cover plate 33.

Meanwhile, in other embodiments of the present application, the number of the guide rails may be two, the two guide rails are respectively provided on the two opposite chamber sidewalls 112 of the sample processing chamber, the insertion of the partition 20 is guided by the two guide rails at the same time, and the partition 20 can be clamped by the two guide rails, so that the width of the partition 20 may be set to be equal to or smaller than the space between the chamber sidewalls 112 on both sides in the width direction thereof, compared to the previous embodiments of the present application, so that the installation is more convenient.

In some embodiments of the present application, a cracking aid for cracking the sample to be processed is pre-stored in the cracking chamber 11, and is cracked into a cracking process, and/or a cleaning reagent for cleaning the sample to be processed is pre-stored in the cleaning chamber 12, and is cleaned into a cleaning process.

The cracking aid is the cracking particle provided in the previous embodiment, and may be polystyrene particles, borosilicate glass particles, quartz glass particles, etc. The cracking chamber 11 is a cracking chamber, and the cleaning chamber 12 is a cleaning chamber.

In these embodiments of the present application, by providing the partition plate 20 in the cleavage chamber 11 as the cleavage chamber and dividing the cleavage chamber 11 into the first region 102 and the second region 103 in which the bottom regions communicate with each other and the corresponding regions of the partition plate 20 are isolated from each other. The amount of the lipophilic substance 40 charged into the lysis chamber 11 can be reduced, and the accommodating space of the second region 103 can be enlarged to accommodate the sample adding member 30.

In some embodiments of the application, the bottom of the partition 20 is spaced a predetermined distance from the bottom of the lysis chamber 11 such that the bottom regions of the first region 102 and the second region 103 are in communication with each other.

Based on this, by providing a gap between the bottom of the partition 20 and the cleavage cavity 11, i.e., the first region 102 is in complete communication with the bottom end of the second region 103, the amount of sample to be treated that can be injected into the cleavage cavity 11 is not affected and movement of the magnetically permeable member 50 between the first region 102 and the second region 103 is facilitated.

The preset distance may be set according to practical situations on the premise of ensuring that the liquid level of the detection liquid contained in the cleavage chamber 11 is higher than the bottom end of the partition plate 20.

In a second aspect, embodiments of the present application provide a biological detection system comprising a biological sample processing device 100 and a magnet, and/or a heating device, and/or an ultrasound device of any of the previous embodiments. Wherein the magnet may abut the outer chamber side wall of the sample processing member 10 and be adapted to provide an external magnetic field; the heating device is abutted against the side wall of the outer chamber of the communicating channel 13 and/or the cleaning cavity 12 and is used for heating; the ultrasound device abuts against the outer chamber side wall of the lysis chamber 11 and is adapted to emit ultrasound waves towards the lysis chamber 11 and/or the communication channel 13.

Fig. 7 is a schematic illustration of a biological sample processing device according to some embodiments of the present application, and fig. 6 is a schematic illustration of a prior art, in which a sample 400 to be processed may be pre-packaged in the biological sample processing device before the biological sample processing device 100 is manufactured, or may be added by a user during the testing process. In either case, however, the amount of the sample 400 to be processed is maintained at a range value for the accuracy of the detection result, and the lysis chamber 11 is not preferably provided too long, so that the cross-sectional area of the lysis chamber 11 is relatively fixed. In this case, since a sufficient amount of paraffin 300 is required to be packed during the movement of the magnetic particles to ensure the loss of the magnetic beads, the paraffin 300 in the liquid form needs to cover the entire cross section of the pyrolysis chamber 11 as shown in fig. 6, resulting in a large amount of paraffin 300, which does not contribute to saving materials and rapid melting. By adopting the scheme shown in fig. 7 in this embodiment, the partition plate 20 is provided, so that the liquid paraffin 300 entering the cracking chamber 11 later can be partitioned into a smaller area without increasing additional cost, and the amount of paraffin 300 is reduced and sufficient paraffin wrapping of the magnetic particles in the moving process can be ensured as well on the necessary path of the magnetic particles in the moving process. Thereby ensuring the detection efficiency and the accuracy of the detection result, reducing the overall size of the biological sample processing device 100, and further ensuring the convenience of transportation, storage and use by users.

Although the present application is not limited to the embodiments, those skilled in the art will readily appreciate that various modifications and substitutions are possible, and these are within the scope of the embodiments disclosed herein. Therefore, the protection scope of the embodiments of the present application shall be subject to the protection scope of the claims.

Claims

1. A biological sample processing device, comprising:

a sample processing member;

the device comprises a cracking cavity for receiving a sample to be treated and cracking the sample to be treated;

the device comprises a cleaning cavity which is arranged side by side with the cracking cavity and is used for cleaning a sample to be processed;

and a communication channel for communicating the top of the cracking chamber and the cleaning chamber;

the lysis cavity is used for containing an aqueous solution containing a sample to be treated;

the separation plate is arranged in the cracking cavity, the separation plate divides the cracking cavity into a first area and a second area, the bottom areas of the first area are communicated with each other, the corresponding areas of the separation plate are isolated from each other, and the first area is positioned between the communication channel and the second area;

When the lysis cavity contains an aqueous solution containing a sample to be treated, one end of the baffle plate stretches into the lower part of the liquid surface of the aqueous solution;

a lipophilic substance which can change shape along with the change of temperature is also arranged between the first area and the tail end of the communication channel; the lipophilic substance isolates the liquid flow of the cracking cavity and the cleaning cavity in a solid state at normal temperature; the lipophilic material is overlaid on top of the aqueous solution in liquid form, the lipophilic material being immiscible with the aqueous solution.

2. The biological sample processing device of claim 1, further comprising a magnetically permeable member received in at least one of the lysis chamber, the wash chamber, and the communication channel, the magnetically permeable member configured to enter the lysis chamber and adsorb a target substance in a sample to be processed under the influence of an external magnetic field.

3. The biological sample processing device of claim 2, wherein said lipophilic substance is solid at a first temperature and is at least partially contained within said communication channel, said magnetically permeable member being immobilized within said lipophilic substance.

4. A biological sample processing device according to claim 3, wherein the lipophilic substance is meltable at a second temperature to flow into the first region and float above the sample to be processed contained in the lysis chamber, the second temperature being greater than the first temperature.

5. The biological sample processing device according to claim 4, wherein said magnetically permeable member within said communication channel is adapted to pass through said lipophilic substance into said lysis chamber and capture a target substance in contact with a sample to be processed and through said lipophilic substance to said wash chamber under the influence of an external magnetic field.

6. The biological sample processing device of claim 5, wherein said communication channel comprises a first port adjacent to said lysing chamber and a second port adjacent to said wash chamber;

7. The biological sample processing device of claim 1, wherein the lysis chamber comprises a chamber bottom wall, a plurality of chamber side walls formed by bending and extending from the chamber bottom wall, the chamber side walls comprising a first region side wall and a second region side wall disposed in spaced relation to the partition, the first region side wall being adjacent to the communication channel, a spacing between the partition and the first region side wall being less than a spacing between the partition and the second region side wall.

8. The biological sample processing device of claim 1, further comprising a loading member at least partially inserted into the second region, the loading member having a receiving cavity for receiving a sample to be processed and a filter outlet communicating the receiving cavity with the second region.

9. The biological sample processing device according to claim 8, wherein the communication channel, the lysis chamber and the washing chamber have openings in the same direction, and the sample addition member further comprises a cover plate covering the openings;

the cover plate and the partition plate are arranged at intervals;

or, the cover plate is abutted with the partition plate.

10. The biological sample processing device of claim 8, wherein said sample processing member further comprises an opening communicating with said communication channel and said lysis chamber and said wash chamber, said sample application member further comprises a cover plate covering said opening, and said partition plate is connected to said cover plate.

11. The biological sample processing device of claim 7, wherein the lysis chamber comprises a chamber bottom wall and a plurality of chamber side walls formed by bending and extending from the chamber bottom wall, and wherein the partition is in interference fit with the chamber side walls in the width direction thereof.

12. The biological sample processing device of claim 7, wherein the lysis chamber comprises a chamber bottom wall, a plurality of chamber side walls extending from the chamber bottom wall in a folded configuration, and a rail extending inwardly from the chamber side walls, the partition being captured in the rail.

13. The biological sample processing device of claim 1, wherein a crack aid for breaking the sample to be processed is pre-stored in the crack chamber.

14. The biological sample processing device of claim 1, wherein a predetermined distance is provided between the bottom of the partition and the bottom of the lysis chamber to allow the bottom regions of the first and second regions to communicate with each other.

15. A biological detection system, comprising:

the biological sample processing device of any one of claims 1-14; and

The magnet is abutted against the side wall of the outer chamber of the sample processing component and is used for providing an external magnetic field;

the heating device is abutted against the side wall of the outer cavity of the communication channel or the cleaning cavity and is used for heating;

the ultrasonic device is abutted to the side wall of the outer chamber of the cracking cavity and used for transmitting ultrasonic waves to the cracking cavity or the communication channel.