WO2024049165A1

WO2024049165A1 - Sample pretreatment device

Info

Publication number: WO2024049165A1
Application number: PCT/KR2023/012796
Authority: WO
Inventors: 이기자; 안병기
Original assignee: 주식회사 덴오믹스
Priority date: 2022-08-29
Filing date: 2023-08-29
Publication date: 2024-03-07
Also published as: KR20240029997A

Abstract

The present invention provides a sample pretreatment device comprising: a pretreatment-dedicated container part filled with a mixture of a bacterial solution and a sample introduced for bacterial testing; a rotating part on which a container body of the pretreatment-dedicated container part is mounted and which rotates the mounted container body to simultaneously stimulate and uniformly mix the mixture; a heat treatment part which is provided spaced apart from the rotating part and heats the container body to heat-treat the mixture filled in the container body; a power supply unit which is connected to the rotating part and the heat treatment part and supplies external power to the rotating part and the heat treatment part; and an operation unit which is connected to the rotating part, the heat treatment part, and the power supply unit, and operated by a user to control the operation of the rotating part, the heat treatment part, and the power supply unit. Accordingly, the present invention can be used for the quick and efficient pretreatment of a mixture during disease diagnosis using pathogenic bacterial DNA, and can be used for the early diagnosis and preventive management of not only oral diseases such as periodontitis and gingivitis, but also various systemic diseases linked to oral health, through rapid bacterial testing enabling rapid qPCR analysis.

Description

Sample preparation device

The present invention relates to a sample pretreatment device, and more specifically, to a sample pretreatment device that allows rapid quantitative testing of oral bacteria in one step.

[National research and development project that supported this invention]

[Assignment number] 1711137928

[Assignment number] KMDF_PR_20200901_0023

[Ministry Name] Ministry of Science and ICT, Ministry of Trade, Industry and Energy, Ministry of Health and Welfare, Ministry of Food and Drug Safety

[Project management (professional) organization name] Pan-Ministerial Medical Device Research and Development Project Group

[Research project name] Pan-ministry full-cycle medical device research and development project

[Research project name] Development of saliva-based on-site pretreatment module and oral pathogenic bacteria rapid test kit

[Contribution rate] 1/1

[Name of project carrying out organization] Kyunghee University Industry-Academic Cooperation Foundation

[Research period] 2020.09.01 ~ 2024.12.31

The oral microbiome refers to the microorganisms present in the oral cavity and their entire genetic information. It is the most common oral disease and infectious disease when the ecological balance of the oral microbiota is shifted to an imbalanced state or when the complexity of the oral microbiota increases. Dental caries and periodontal disease, which are diseases, occur. In addition, periodontal disease is known to increase the risk of developing systemic diseases such as diabetes, cardiovascular disease, and dementia, and in the case of rheumatoid arthritis, the risk of developing rheumatoid arthritis has been reported to increase by up to 1.93 times. This is known to be because Porphyromonas gingivalis , a representative Gram-negative anaerobic bacterium that causes periodontitis, secretes peptidylarginine deiminase (PADs), which contributes to the production of anti-CCP antibodies (anti-cyclic citrullinated peptide Ab) in rheumatoid arthritis. Therefore, accurate analysis of oral bacteria can be of great help not only in the diagnosis of oral diseases but also in the prevention and management of systemic diseases.

Oral bacterial testing can be largely divided into sample preparation and real-time PCR analysis. While real-time PCR analysis is performed on a device, in the case of sample pretreatment, the complex bacterial DNA extraction process must be performed by professional personnel and the device and A labor-intensive process is necessary because several complex processes must be carried out using instruments.

The technology behind the present invention is disclosed in Korean Patent No. 10-1467371 (registered on November 25, 2014).

The present invention was created to meet the above-mentioned situation, and uses a mixed solution in which a sample for bacterial testing is added to a bacterial lysate to perform one simple pretreatment for pathogenic bacteria present in the sample without a complex bacterial DNA extraction process. The purpose is to provide a sample preparation device that can perform qPCR analysis immediately after processing.

In order to achieve the above object, the present invention includes a pretreatment-only container filled with a mixed solution containing a bacterial solution mixed with a sample introduced for bacterial testing; A rotating unit on which the container body of the pretreatment dedicated container unit is mounted and rotates the mounted container body to mix the mixed solution evenly and stimulate it at the same time; a heat treatment unit that is provided to be spaced apart from the rotating unit and heats the container body to heat-treat the mixed liquid filled in the container body; a power supply unit connected to the rotating unit and the heat treatment unit and supplying external power to the rotating unit and the heat treatment unit; and a control unit connected to the rotating unit, the heat treatment unit, and the power supply unit, and operated by a user to control operations of the rotating unit, the heat treatment unit, and the power supply unit.

The sample pretreatment device according to the present invention may further include a case portion in which the rotation portion, the heat treatment portion, the power supply portion, and the manipulation portion are spaced apart from each other.

In the sample pretreatment device according to the present invention, the pretreatment-only container portion has a cylindrical shape that is closed at one end and open at the other end, the interior of which is filled with a bacterial solution, and a container body provided with an airtight lid that seals the open other end; A sample funnel is attached and detachable to the other open end of the container body and collects a sample into the interior of the container body, and when the sample funnel is detached from the container body, one end is mounted on the other open end of the container body, The container body may include an extraction cap formed at the other end of an extraction port for extracting a mixed solution of bacterial lysate and a sample from the container body, and a filter for filtering out foreign substances in the mixed solution inside the extraction cap. It can be provided.

The pretreatment-only container part is mounted on the outlet of the extraction cap and may further include a discharge tip that discharges a small amount of mixed liquid through the outlet when the extraction cap is pressed. Inside the discharge tip, the liquid that has passed through the filter is disposed. A syringe filter that filters out smaller-sized foreign substances may be provided.

The extraction cap may include a main body having a cylindrical shape, and a cover mounted on one end of the main body and provided with the extraction port for extracting the mixed liquid, and a seating protrusion on which the filter is seated may be formed on the inside of the main body. The filter seated on the seating protrusion may be fixed within the main body when the cover is mounted on the main body, and the other end of the main body and the other end of the container body may be fastened by a fitting method.

The sample funnel includes an insertion end having a cylindrical shape and one side of which is inserted into the other open end of the container body, and a collection end that protrudes obliquely outward from the other side of the insertion end and collects the sample supplied into the interior of the container body. can do.

The bacterial lysate filled inside the container body may include Tris-HCl, NaCl, SDS, Nonidet P-40, and Chelex-100 resin, and the sample may be saliva.

The rotating part includes a mounting body on which the container body of the pretreatment dedicated container portion is mounted, a rotating motor connected to the mounting body and a rotating shaft and providing power to rotate the mounting body, and a support frame supporting the rotating motor, It may include a rotation speed regulator that adjusts the rotation speed of the rotation motor.

The mounting body is connected to the rotating shaft in a disk shape, and a mounting groove in which the container body is mounted is formed in the central portion, a mounting protrusion is formed on the circumferential surface, and a first mounting body to which a first magnet is attached is attached to the inside of the circumferential surface. It has a disk shape, and a through hole through which the container body passes is formed in the central portion, a coupling groove coupled to the mounting protrusion is formed on the circumferential surface, and a second magnet magnetically attached to the first magnet is formed on the inside of the circumferential surface. It may include a second mounting body to which two magnets are attached.

The rotating part may be provided at a lower portion of the support frame and may further include a slide rail that guides the support frame to slide toward the heat treatment unit, and a wheel member that moves on the slide rail may be provided on the lower surface of the support frame. It can be.

The heat treatment unit includes a heat treatment body into which the container body slidably moved by the rotation unit is inserted, a ceramic heater provided on the heat treatment body and applying heat to the container body inserted into the heat treatment body, and the ceramic heater. It is connected to and may include a temperature controller that controls the temperature of the ceramic heater.

The heat treatment unit may heat treat the mixed liquid filled inside the container body at a temperature of 95 degrees for more than 10 minutes or at a temperature of 70 degrees for more than 20 minutes.

The operating unit is connected to the power supply unit and includes an on/off switch that operates the power supply unit to turn on/off the operation of the rotating unit and the heat treatment unit, and is connected to the rotation speed controller to set the rotation speed of the rotation motor. It may include a first operation button that is connected to the temperature controller and operated to set the temperature of the ceramic heater.

According to the sample pretreatment device according to the present invention, it can be used for quick and efficient pretreatment of mixed solutions in disease diagnosis using pathogenic bacterial DNA, and can be used for rapid bacterial testing capable of qPCR analysis within a short period of time, as well as oral diseases such as periodontitis and gingivitis. It can also be used for early diagnosis and preventive management of various systemic diseases that are linked to oral health.

Figure 1 is a diagram illustrating a sample pretreatment device according to an embodiment of the present invention.

FIG. 2 is a diagram illustrating the basic configuration of a preprocessing-only container used in the sample pretreatment device shown in FIG. 1 to collect samples.

Figure 3 is a diagram schematically showing a state in which a discharge tip for discharging a small amount of mixed liquid is mounted on the extraction cap shown in Figure 2.

Figure 4 is a diagram schematically showing the process of collecting a sample and extracting a mixed solution using the pre-treatment dedicated container shown in Figures 2 and 3.

Figure 5 is a diagram schematically showing a process in which the container body shown in Figure 2 is mounted on the mounting body of the rotating part.

FIG. 6 is a diagram schematically showing the process in which the container body is moved to the heat treatment unit by the rotating unit shown in FIG. 1.

Hereinafter, preferred embodiments according to the present invention will be described in detail with reference to the attached drawings. Prior to this, the terms or words used in this specification and claims should not be construed as limited to their usual or dictionary meanings, and the inventor should appropriately define the concept of terms in order to explain his or her invention in the best way. It must be interpreted as meaning and concept consistent with the technical idea of the present invention based on the principle of definability.

Referring to Figures 1 to 6, the sample pretreatment device 100 according to an embodiment of the present invention is designed to enable rapid quantitative testing of oral bacteria in one step, and includes a container unit 1100 dedicated to pretreatment, It includes a rotating unit 1200, a heat treatment unit 1300, a power supply unit 1400, and an operating unit 1500, and may further include a case unit 1600.

Referring to Figures 2 to 4, the interior of the pretreatment-only container unit 1100 is filled with a mixed solution in which a bacterial solution is mixed with a sample introduced for bacterial testing, and the pretreatment-only container unit 1100 is a container body ( 1110), a sample funnel 1120, an extraction cap 1130, and may further include a discharge tip 1140.

The container body 1110 is cylindrical in shape, one end of which is closed and the other end open, the inside is filled with a bacterial solution through the open other end, and the open other end is provided with an airtight lid 1111 for sealing, The airtight lid 1111 and the container body 1110 are preferably connected by a connecting member 1112.

A sample funnel 1120 is attached and detachable to the other open end of the container body 1110, and the sample funnel 1120 is used to collect samples for performing a rapid quantitative test for oral bacteria inside the container body 1110. It is attached to the container body 1110 for this purpose.

The sample funnel 1120 includes an insertion end 1121 and a collection end 1122, and the insertion end 1121 has a cylindrical shape and one side is inserted into the other open end of the container body 1110. The diameter D1 of the insertion end 1121 is preferably the same as or smaller than the inner diameter D2 of the other open end of the container body 1110.

On the other side of the insertion end 1121, a collection end 1122 protrudes obliquely outward, and the collection end 1122 serves to collect the sample supplied into the container body 1110, and the sample funnel Through 1120, the sample can be easily collected into the interior of the container body 1110.

The bacterial lysate filled inside the container body 1110 includes Tris-HCl, NaCl, SDS, Nonidet P-40, and Chelex-100 resin, and the sample introduced into the bacterial lysate is preferably saliva. do.

Referring to Figures 1 to 4, the sample funnel 1120 is attached to the container body 1110 only when collecting a sample, and is detached from the container body 1110 when sample collection is completed. In order to pre-process the mixed solution in which the sample introduced for bacterial testing is mixed with the bacterial solution filled inside (1110), the open other end of the container body 1110 is sealed by the airtight lid 1111 and then sealed. The container body 1110 is mounted on the rotating unit 1200.

The mixed solution is evenly mixed and stimulated by the rotating unit 1200, and the mixed solution heat-treated in the heat treatment unit 1300, which will be described later, is extracted from the container body 1110. When the mixed solution is extracted from the container body 1110, the extraction cap ( 1130) is used, and when extracting a smaller amount, the discharge tip 1140 is used.

The extraction cap 1130 has one end mounted on the other open end of the container body 1110 when extracting the mixed liquid from the container body 1110, and the extraction cap 1130 is connected to the body 1131 and the cover ( 1132) and a filter 1133. The main body 1131 has a cylindrical shape, and a cover 1132 is installed at one end of the main body 1131 with an extraction port 1132a through which a mixture of bacterial lysate and sample is extracted, and the cover ( 1132) and the main body 1131 are preferably connected by a connecting string 1132a. The container body 1110 is made of hard polypropylene material to facilitate heat treatment in the heat treatment unit 1300, which will be described later, and the extraction cap 1130 is made of soft polychloride material to facilitate coupling with the container body 1110. It is preferably made of vinyl material, and the main body 1131 of the extraction cap 1130 and the container main body 1110 are coupled by a fitting method, and the diameter D3 of one end of the main body 1131 is the container. It is preferable that the inner diameter D2 of the other open end of the main body 1110 is the same.

A filter 1133 is provided inside the main body 1131, and the filter 1133 filters so that foreign substances in the mixed solution are not discharged through the extraction cap 113 when the mixed solution is discharged through the extraction cap 1130. It plays a role. A seating protrusion 1131a on which the filter 1133 is seated is formed inside the main body 1131, and the filter 1133 seated on the seating protrusion 1131a is formed by the cover 1132 of the main body 1131. ), it is fixed within the main body 1131.

A discharge tip 1140 is mounted on the extraction port 1132a of the extraction cap 1130, and the discharge tip 1140 is connected to the extraction port 1132a when an operator presses the extraction cap 1130 to discharge the mixed liquid. ), which discharges the mixed solution drop by drop in small amounts (7 to 8 μL).

The container body 1110, sealed by the airtight lid 1111, is mounted on a rotating unit 1200 for pretreatment of the mixed liquid, and the rotating unit 1200 rotates the mounted container body 1110 to fill the mixed liquid. It plays a stimulating role by mixing well.

The rotating unit 1200 includes a mounting body 1210, a rotating motor 1220, a support frame 1230, and a rotating speed controller 1240, and may further include a slide rail 1250.

The container body 1110 sealed by the airtight lid 111 is mounted on the mounting body 1210, and the mounting body 1210 rotates by a rotation motor 1220 connected to the rotating shaft 1221. The mixed liquid in the container body 1110 is mixed well and stimulated. The rotation motor 1220, which provides power to rotate the mounting body 1210 by the rotation shaft 1221, is general, and detailed description thereof will be omitted. The rotation motor 1220 preferably rotates the container body 1110 at 12 to 30 rpm for 10 minutes to apply physical stimulation to the mixed solution, but is not limited to this and rotates for 10 minutes depending on the intensity of the applied physical stimulation. It is desirable that the number of rotations is increased or decreased.

Referring to FIG. 5, the mounting body 1210 on which the container body 1110 is mounted includes a first mounting body 1211 and a second mounting body 1212. The first mounting body 1211 is connected to the rotating shaft 1221 in a disk shape, and a mounting groove 1211a on which the container body 1110 is mounted is formed in the central portion, and a mounting protrusion 1211b is formed on the outer circumferential surface. It is formed to protrude, and a first magnet 1211c is attached to the inside of the circumferential surface.

The second mounting body 1212 coupled to the first mounting body 1211 has the same disk shape as the first mounting body 1211, and has a through hole 1212a in the central portion through which the container body 1110 passes. ) is formed, and a coupling groove (1212b) coupled to the mounting protrusion (1211b) is formed on the circumferential surface, and a second magnet (1212c) is attached to the first magnet (1211c) by magnetic force on the inside of the circumferential surface. ) is attached.

Referring to Figures 5 and 6, the rotation motor 1220, which provides power to rotate the mounting body 1210, is supported by a support frame 1230, and a slide rail is provided at the bottom of the support frame 1230. 1240 is provided, and the slide rail 1240 serves to slide and move the support frame 1230 toward the heat treatment unit 1300. As the support frame 1230 moves toward the heat treatment unit 1300, the container body 1110 mounted on the mounting body 1210 rotated by the rotation motor 1220 is moved within the heat treatment unit 1300. It is located. The rotational speed of the rotation motor 1220, which provides power to rotate the mounting body 1210, is controlled by the rotation speed controller 1240, and the rotation speed controller 1240 is connected to the operation unit 1500, which will be described later. It is desirable to be

The support frame 1230 is slid and moved on the slide rail 1240 by an operator applying external force to the support frame 1230, and the lower surface of the support frame 1230 is moved by the slide rail 1240. It is preferable that the support frame 1230 is provided with moving wheels (not shown) so that it can be easily moved.

1 and 6, a heat treatment unit 1300 is provided adjacent to the rotating unit 1200, and the heat treatment unit 1300 heats the container body 1110 from the outside to heat the container body 1110. It serves to heat treat the mixed liquid inside.

The heat treatment unit 1300 includes a heat treatment body 1310, a ceramic heater 1320, and a temperature controller 1330. The heat treatment body 1310 surrounds the container body 1110, which is slidably moved by the rotating part 1200, from the outside, and the heat treatment body 1310 is provided with the ceramic heater 1320. The ceramic heater 1320 serves to apply heat to the container body 1110 inserted into the heat treatment body 1310.

The ceramic heater 1320 is connected to a temperature controller 1330, and the temperature controller 1330 serves to control the temperature of the ceramic heater 1320 that heats and heat-treats the container body 1110.

The temperature controller 1330 allows the user to easily set the temperature of the ceramic heater 1320 by manipulating the operation unit 1500, which will be described later, and the heat treatment unit 1300 is a heat treatment unit 1300 filled inside the container body 1110. It is preferable to heat-treat the mixed solution at a temperature of 95 degrees for 10 minutes or more or at a temperature of 70 degrees for 20 minutes or more.

The rotating unit 1200 and the heat treatment unit 1300 are connected to a power supply unit 1400, and the power supply unit 1400 supplies external power to the rotating unit 1200 and the heat treatment unit 1300. It plays a role.

The rotation unit 1200, the heat treatment unit 1300, and the power supply unit 1400 are connected to the operation unit 1500, and the operation unit 1500 is connected to the rotation unit 1200, the heat treatment unit 1300, and the power supply unit. The user operates to control the operation of (1400), and the rotating unit (1200), the heat treatment unit (1300), the power supply unit (1400), and the operating unit (1500) are provided in the case unit (1600). desirable.

The manipulation unit 1500 includes an on-off switch 1510, a first manipulation button 1520, and a second manipulation button 1530, and the on-off switch 1510 is connected to the power supply unit 1400. do. By pressing the on-off switch 1510 to operate or stop the power supply unit 1400, the rotation unit 1200 and the heat treatment unit 1300 are operated or stopped.

The rotation speed controller 1240 of the rotation unit 1200 is connected to the first operation button 1520, and the rotation speed of the rotation motor 1220 is easily set by manipulating the first operation button 1520. The temperature of the ceramic heater 1320 can be easily set by operating the second operation button 1530 connected to the temperature controller 1330 of the heat treatment unit 1300.

In order to evaluate the performance of the pretreatment-only container unit 1100, which is an experimental group used in the sample pretreatment device 100 that can recover pretreated samples without using centrifugation according to an embodiment of the present invention, centrifugation was used. The pretreated samples were compared with the recovered control group.

First, in a container body containing 100 μL of a 10-fold concentrated bacterial lysate (ingredients: Tris-HCl, NaCl, SDS, Nonidet P-40, Chelex-100), a S. mutans bacterial solution (1) prepared from artificial saliva was added. x 10 ⁷ CFU/mL) was added and then reacted at 70°C for 20 minutes while operating the rotary motor at a speed of 30 rpm using the one-step sample preparation device 100 according to the present invention.

After attaching the extraction cap to the container body where pretreatment was completed, 100 μL of the sample solution was recovered in a new tube by pressing it by hand (experimental group). 5 μL of this was used as a template for qPCR and S. mutans -specific primer-probe. qPCR was performed using -probe, PP). At this time, add 10 μL of 2X master mix, 2 μL of S. mutans PP, and 3 μL of DW to make a final volume of 20 μL. qPCR conditions were initial denaturation (95°C; 10 minutes - 1 cycle) followed by 2-step PCR conditions (denaturation at 95°C; 15 seconds; binding & elongation at 60°C; 1 minute - 45 cycles). S. mutans DNA was used as a positive control for qPCR, and DW was used as a negative control.

In the case of the control group, the container body with completed pretreatment was centrifuged (12000 rpm, 10 minutes), then 5 μL of the supernatant was taken and used as a template for qPCR, and qPCR was performed in the same way as the experimental group.

As a result, the experimental group showed similar Ct values to the control group as shown in [Table 1] below, so it was confirmed that sample recovery was possible using a pretreatment dedicated container equipped with a filter without using centrifugation.

In addition, in order to evaluate the performance of the sample preparation device 100 according to the present invention, qPCR was measured on samples pretreated with an optimal protocol using a commercially available reagent, one-step DNA/RNA extraction buffer (CHAI, control). It was compared with the Ct value of . Performance evaluation was conducted on S. mutans and P. gingivalis , which are bacterial biomarker candidates for periodontitis diagnosis.

First, in a container containing 100 μL of a 10-fold concentrated bacterial lysate (ingredients: Tris-HCl, NaCl, SDS, Nonidet P-40, Chelex-100), S. mutans bacteria with three different numbers of bacteria were placed in the container body. After adding 1 mL of each solution [100-, 500-, and 1000-fold diluted solution (based on stock vial)], the mixture was reacted at 70°C for 20 minutes using a sample preparation device according to the present invention.

After the pretreatment was completed, the extraction cap was connected to the container body, and 100 μL of the sample solution was collected in a new tube. 5 μL of this was used as a template for qPCR, and qPCR was performed using S. mutans PP.

In the case of the control group, according to the manufacturer's manual, 1 mL of S. mutans bacterial solution (solution diluted 100, 500, and 1000 times the same as the experimental group) with three different bacterial counts was added to each 1.5 mL microtube. Add 100 μL of commercial reagent, vortex for 15 seconds, and react at room temperature for 15 minutes. Next, it was placed in a heating block heated to 60 degrees, treated for 5 minutes, centrifuged at 12000 rpm for 10 minutes, and 100 μL of the sample solution was recovered in a new tube using a micropipette. Of these, 5 μL was used as a template for qPCR, and qPCR was performed using S. mutans PP. At this time, qPCR conditions were the same as the experimental group.

For P. gingivalis , the sample solution was recovered in the same manner using the sample preparation device and dedicated container according to the present invention, 5 μL of this was used as a template for qPCR, and qPCR was performed using P. gingivalis- specific PP. did. In the case of the control group, the procedure was the same as above.

As a result, in all bacterial solutions with three different numbers of bacteria for four types of bacteria, those pretreated with the sample preparation device according to the present invention showed lower Ct values compared to commercially available reagents, as shown in [Table 2]. , it was confirmed that preprocessing performance was superior.

Therefore, in the diagnosis of diseases using pathogenic bacterial DNA, it can be used for quick and efficient pretreatment of mixed solutions, and through rapid bacterial tests that enable qPCR analysis within a short period of time, not only oral diseases such as periodontitis and gingivitis, but also various systemic diseases linked to oral health. It can also be used for early diagnosis and preventive management.

The present invention has been described with reference to the embodiments shown in the drawings, but these are merely illustrative, and those skilled in the art will understand that various modifications and equivalent other embodiments are possible therefrom. Therefore, the true scope of technical protection of the present invention should be determined by the technical spirit of the attached patent claims.

The present invention can be applied to the field of oral bacteria testing.

Claims

A pre-treatment container portion filled with a mixture of the bacterial solution and the sample introduced for bacterial testing;

A rotating unit on which the container body of the pretreatment dedicated container unit is mounted and rotates the mounted container body to mix the mixed solution evenly and stimulate it at the same time;

a heat treatment unit that is provided to be spaced apart from the rotating unit and heats the container body to heat-treat the mixed liquid filled in the container body;

a power supply unit connected to the rotating unit and the heat treatment unit and supplying external power to the rotating unit and the heat treatment unit; and

A sample pretreatment device that is connected to the rotating unit, the heat treatment unit, and the power supply unit, and includes a control unit operated by a user to control operations of the rotating unit, the heat treatment unit, and the power supply unit.
According to claim 1,

A sample pretreatment device further comprising a case portion in which the rotation portion, the heat treatment portion, the power supply portion, and the manipulation portion are spaced apart from each other.
According to claim 1,

The pre-treatment dedicated container part,

A container body having a cylindrical shape that is sealed at one end and open at the other end, filled with a bacterial solution inside, and equipped with an airtight lid that seals the open other end;

A sample funnel that is attached and detachable to the other open end of the container body and collects the sample into the interior of the container body;

When the sample funnel is detached from the container body, one end is mounted on the other open end of the container body, and an extraction port for extracting a mixture of bacterial lysate and sample from the container body is located at the other end. It includes an extraction cap formed,

A sample pretreatment device equipped with a filter inside the extraction cap to filter out foreign substances in the mixed solution.
According to clause 3,

The pre-treatment dedicated container part,

It is mounted on the outlet of the extraction cap and further includes a discharge tip that discharges a small amount of mixed liquid through the outlet when the extraction cap is pressed,

A sample pretreatment device provided inside the discharge tip with a syringe filter that filters out smaller-sized foreign substances that pass through the filter.
According to clause 4,

The extraction cap is,

A main body having a cylindrical shape,

It is mounted on one end of the main body and includes a cover provided with the extraction port for extracting the mixed liquid,

A seating protrusion on which the filter is seated is formed on the inside of the main body,

The filter seated on the seating protrusion is fixed within the main body when the cover is mounted on the main body,

A sample pretreatment device, characterized in that the other end of the main body and the other end of the container body are fastened by a fitting method.
According to clause 3,

The sample funnel is,

an insertion end having a cylindrical shape and one side of which is inserted into the other open end of the container body;

A sample pretreatment device including a collection end that protrudes obliquely outward from the other side of the insertion end and collects the sample supplied into the container body.
According to clause 3,

The bacterial solution filled inside the container body includes Tris-HCl, NaCl, SDS, Nonidet P-40, and Chelex-100 resin,

A sample pretreatment device, characterized in that the sample is saliva.
According to claim 1,

The rotating part,

A mounting body on which the container body of the pre-processing dedicated container portion is mounted,

A rotation motor connected to the mounting body by a rotating shaft and providing power to rotate the mounting body,

A support frame supporting the rotation motor,

A sample pretreatment device including a rotation speed controller that controls the rotation speed of the rotation motor.
According to clause 8,

The mounting body is,

A first mounting body that is connected to the rotating shaft in a disk shape and has a mounting groove in the center where the container body is mounted, a mounting protrusion is formed on the circumferential surface, and a first magnet is attached to the inside of the circumferential surface;

A through hole through which the container body penetrates is formed in the central portion of the disc shape, a coupling groove coupled to the mounting protrusion is formed on the circumferential surface, and a second magnet is magnetically attached to the first magnet on the inside of the circumferential surface. A sample pretreatment device including a second mounting body to which this is attached.
According to clause 8,

The rotating part,

It is provided at the lower part of the support frame and further includes a slide rail that guides the support frame to slide toward the heat treatment unit,

A sample pretreatment device wherein a wheel member that moves on the slide rail is provided on the lower surface of the support frame.
According to claim 10,

The heat treatment unit,

a heat treatment body into which the container body slidably moved by the rotating unit is inserted;

a ceramic heater provided in the heat treatment body and applying heat to the container body inserted into the heat treatment body;

A sample pretreatment device connected to the ceramic heater and including a temperature controller that controls the temperature of the ceramic heater.
According to claim 10,

The heat treatment unit heat-treats the mixed solution filled inside the container body at a temperature of 95 degrees for more than 10 minutes or at a temperature of 70 degrees for more than 20 minutes.
According to claim 11,

The control unit,

An on-off switch connected to the power supply unit and turning the rotation unit and the heat treatment unit on/off by operating the power supply unit,

A first operation button connected to the rotation speed controller and operated to set the rotation speed of the rotation motor,

A sample pretreatment device including a second operation button connected to the temperature controller and operated to set the temperature of the ceramic heater.