WO2016117726A1

WO2016117726A1 - Cartridge

Info

Publication number: WO2016117726A1
Application number: PCT/KR2015/000694
Authority: WO
Inventors: Byeong Woo Bae; In Kyoung Hwang; Seong Min Park; Eun Ja Kim
Original assignee: Infopia Co., Ltd.
Priority date: 2015-01-23
Filing date: 2015-01-23
Publication date: 2016-07-28

Abstract

A cartridge includes a plurality of reagent chambers, a discard chamber, a PCR chamber, and a piston provided with a channel in which a first path and a second path are provided, wherein the first path is used to move a reagent between the plurality of reagent chambers, and the second path is used to move a material in the reagent chamber into the discard chamber or the PCR chamber.

Description

CARTRIDGE

The present invention relates to a cartridge, and more particularly, to a cartridge capable of extracting, cleaning, amplifying and measuring nucleic acid using one apparatus.

A method of detecting a pathogenic organism or the like using a reaction of detecting nucleic acid through extraction and amplification is used for medical and industrial purposes. For this, a process of extracting nucleic acid, a process of amplifying the extracted nucleic acid, and a process of measuring the nucleic acid amplified in the amplification process are needed.

In the related art, each of the processes requires an apparatus, which incurs a large expense, and execution of the processes is time-consuming. In addition, when each of the processes is performed by an individual apparatus, a material may be contaminated upon exchange of the material between the apparatuses.

An object of the present invention is to provide a cartridge capable of extracting, cleaning, amplifying and measuring nucleic acid using one apparatus.

Another object of the present invention is to provide a cartridge capable of reducing a time consumed in nucleic acid extraction, cleaning, amplification and measurement processes.

It will be appreciated by those skilled in the art that the objects of the present invention are not limited to the above-mentioned objects, and objects not described herein will be clearly understood from the specification and the accompanying drawings.

According to an aspect of the present invention, there is provided a cartridge including a plurality of reagent chambers; a discard chamber; a PCR chamber; and a piston provided with a channel in which a first path and a second path are provided, wherein the first path is used to move a reagent between the plurality of reagent chambers, and the second path is used to move a material in the reagent chamber to the discard chamber or the PCR chamber.

It will be appreciated by those skilled in the art that the means of the present invention are not limited to the above-mentioned means, and means not described herein will be clearly understood from the specification and the accompanying drawings.

According to the present invention, a processing time can be reduced by mixing a reagent using a channel of a piston, transmitting the mixed reagent to a filter unit, and extracting and cleaning nucleic acid.

According to the present invention, contamination of a PCR reagent can be prevented by moving the PCR reagent using a separate auxiliary channel.

It will be appreciated by those skilled in the art that the effects of the present invention are not limited to the above-mentioned effects, and effects not described herein will be clearly understood from the specification and the accompanying drawings.

FIG. 1 is a block diagram showing an automatic analysis apparatus according to an embodiment;

FIG. 2 is a perspective view showing a cartridge according to the embodiment;

FIG. 3 is an exploded perspective view showing the cartridge according to the embodiment;

FIG. 4 is a plan view showing a chamber housing according to the embodiment;

FIG. 5 is a cross-sectional view showing the chamber housing according to the embodiment;

FIG. 6 is a cross-sectional view showing a state in which a piston according to the embodiment is lowered;

FIG. 7 is a cross-sectional view showing a lower housing according to the embodiment;

FIG. 8 is a view showing a connecting state of a valve channel according to the embodiment;

FIG. 9 is a view showing a method of controlling an automatic analysis apparatus according to an embodiment;

FIG. 10 is a view showing a process of injecting a sample according to the embodiment;

FIG. 11 is a view showing a process of generating a PK reagent according to the embodiment;

FIG. 12 is a view showing a process of mixing a reagent according to the embodiment;

FIG. 13 is a view showing a process of mixing the reagent according to the embodiment;

FIG. 14 is a view showing the reagent moved to a first path according to the embodiment;

FIG. 15 is a view showing movement of a cleaning reagent according to the embodiment;

FIG. 16 is a view showing movement of an elution reagent according to the embodiment;

FIG. 17 is a view showing a process of generating a PCR reagent according to the embodiment;

FIG. 18 is a view showing a process of generating the PCR reagent according to the embodiment; and

FIG. 19 is a view showing movement of a reagent to an auxiliary channel according to the embodiment.

Since the embodiments disclosed herein are provided to clearly describe the spirit of the present invention to those skilled in the art, the present invention is not limited to the embodiments disclosed herein, and the scope of the present invention should be construed to include changes or modifications without departing from the spirit of the present invention.

The terms used throughout this specification are provided to describe embodiments but not intended to limit the present invention, and may differ due to intentions or customs of those skilled in the art or appearance of new technologies. However, unlike this, when a specified term is defined and used with an arbitrary meaning, the meaning of the term will be separately defined. Accordingly, the terms used herein should be construed based on substantial meaning and contents described throughout the specification rather than simple names of the terms.

The accompanying drawings of the specification are provided to easily describe the present invention and dimensions shown therein may be exaggerated for the purpose of understanding of the present invention, but the present invention is not limited to the drawings.

When it is determined that specific description of known elements or functions related to the present invention may obfuscate the spirit of the present invention, detailed description thereof will be omitted according to necessity.

According to an aspect of the present invention, a cartridge including a plurality of reagent chambers; a discard chamber; a PCR chamber; and a piston provided with a channel in which a first path and a second path are provided, wherein the first path is used to move a reagent between the plurality of reagent chambers, and the second path is used to move a material in the reagent chamber to the discard chamber or the PCR chamber may be provided.

In addition, the channel may be a groove formed in the piston.

In addition, the first path may be provided by rotation of the piston.

In addition, the second path may be provided by vertical movement of the piston.

In addition, the first path may be a path through which a material is moved between the neighboring reagent chambers.

In addition, the cartridge may further include a valve configured to selectively supply the material from the second path into the discard chamber or the PCR chamber.

In addition, the cartridge may further include a filter unit disposed between the second path and the valve and configured to filter a target material from the material from the second path.

In addition, the filter unit may include a membrane filter.

In addition, the cartridge may further include an auxiliary channel configured to connect at least one of the plurality of reagent chambers and the PCR chamber.

In addition, the auxiliary channel may be formed at a lower housing.

In addition, the auxiliary channel may be connected to at least one of the plurality of reagent chambers through the channel.

In addition, a partial region of an outer surface of the PCR chamber may be a light transmission region.

In addition, a pneumatic pressure may be supplied into the plurality of reagent chambers.

In addition, the cartridge may further include a packing member disposed at an outer circumferential surface of a central region of the piston.

In addition, the central region may be an intermediate region between the reagent chamber and the filter unit.

In addition, the valve may include a V-shaped channel.

Hereinafter, an automatic analysis apparatus and a cartridge according to an embodiment of the present invention will be described.

FIG. 1 is a block diagram showing an automatic analysis apparatus according to the embodiment.

Referring to FIG. 1, an automatic analysis apparatus 1 according to the embodiment may include a control unit 10, a power unit 20, a pneumatic unit 30, a temperature adjustment unit 40, and a measurement unit 50.

The automatic analysis apparatus 1 may further include a cartridge 100 configured to perform a nucleic acid extraction and amplification process using the power unit 20, the pneumatic unit 30, the temperature adjustment unit 40 and the measurement unit 50.

The control unit 10 can control the power unit 20, the pneumatic unit 30, the temperature adjustment unit 40 and the measurement unit 50.

The power unit 20 can provide power to the cartridge 100.

The power unit 20 may include a piston power unit 21 and a valve power unit 23. The piston power unit 21 can provide power to a piston of the cartridge 100. The piston power unit 21 can move the piston of the cartridge 100 vertically and can rotate the piston.

The valve power unit 23 can provide power to a valve of the cartridge 100. The valve power unit 23 can rotate the valve of the cartridge 100.

The pneumatic unit 30 can supply a pneumatic pressure into the cartridge 100. The pneumatic unit 30 can selectively supply the pneumatic pressure into a plurality of chambers of the cartridge 100. The pneumatic unit 30 can selectively supply the pneumatic pressure into a plurality of reagent chambers of the cartridge 100. That is, pneumatic unit 30 can selectively inject a gas into the plurality of reagent chambers of the cartridge 100. The pneumatic unit 30 may suction the gas in a discard chamber and a PCR chamber of the cartridge 100.

The temperature adjustment unit 40 can adjust a temperature of a partial region of the cartridge 100. The temperature adjustment unit 40 can adjust a temperature of the PCR chamber of the cartridge 100. The temperature adjustment unit 40 can heat and cool the PCR chamber. The temperature adjustment unit 40 can repeat a process of heating and then cooling the PCR chamber.

The measurement unit 50 can measure whether a specific nucleic acid is present in the amplification process of the nucleic acid accommodated in the PCR chamber. The measurement unit 50 can determine whether the specific nucleic acid is present using a light source. In the amplification process of the nucleic acid accommodated in the PCR chamber, the measurement unit 50 can inject light having a specific wavelength into the PCR chamber to photograph the light discharged through the nucleic acid, and determine whether the specific nucleic acid is present by analyzing the photographed light. While an optical method of measurement by the measurement unit 50 has been exemplarily described, the present invention is not limited thereto.

FIG. 2 is a perspective view showing a cartridge according to the embodiment, and FIG. 3 is an exploded perspective view showing the cartridge according to the embodiment.

Referring to FIGS. 2 and 3, the cartridge 100 according to the embodiment may include a chamber housing 110, an upper cover 120, a piston 130, a packing member 140 and a lower housing 150.

The upper cover 120 is disposed at an upper section of the chamber housing 110, and the lower housing 150 is disposed at a lower section of the chamber housing 110.

A chamber cover 111 may be disposed at an upper surface of the chamber housing 110, and a plurality of injection holes 112 and a chamber through-hole 113 may be formed in the chamber cover 111. The chamber cover 111 may have a shape corresponding to the upper surface of the chamber housing 110. The chamber cover 111 may have a circular shape.

The chamber through-hole 113 may be formed at a central region of the chamber cover 111. The piston 130 may be accommodated through the chamber through-hole 113. A through-hole (not shown) is also formed in the chamber housing 110.

The piston 130 may pass the chamber housing 110 through the through-holes (not shown) of the chamber through-hole 113 and the chamber housing 110. The chamber through-hole 113 may have a shape corresponding to the shape of the piston 130. The chamber through-hole 113 may have a circular shape.

The plurality of injection holes 112 may be formed in a peripheral region of the chamber through-hole 113. The plurality of injection holes 112 may be formed along an outer circumferential surface of the chamber through-hole 113. The plurality of injection holes 112 may be disposed to form a circular shape.

The upper cover 120 may have a shape corresponding to the chamber cover 111. The upper cover 120 may have a circular shape. A plurality of pneumatic pressure injection ports 121, an upper cover through-hole 123 and a sample injection port 125 may be formed in the upper cover 120.

The upper cover through-hole 123 may be formed at a central region of the upper cover 120. The upper cover through-hole 123 may be formed at a position corresponding to the chamber through-hole 113. The piston 130 may be accommodated through the upper cover through-hole 123. The piston 130 may pass through the upper cover through-hole 123.

The plurality of pneumatic pressure injection ports 121 may be formed at a peripheral region of the upper cover through-hole 123. The plurality of pneumatic pressure injection ports 121 may be formed along an outer circumferential surface of the upper cover through-hole 123. The plurality of pneumatic pressure injection ports 121 may be disposed to form a circular shape. The plurality of pneumatic pressure injection ports 121 may be formed at positions corresponding to each of the plurality of injection holes 112. The plurality of pneumatic pressure injection ports 121 can receive the pneumatic pressure from the pneumatic unit 30 to transmit the pneumatic pressure to the injection holes 112. The plurality of pneumatic pressure injection ports 121 may have a fastening structure to be fastened to the pneumatic unit 30.

The sample injection port 125 may be formed to pass through the upper cover 120. The sample injection port 125 may be formed at a region adjacent to any one of the plurality of pneumatic pressure injection ports 121. The sample injection port 125 may come in communication with any one of the plurality of injection holes 112. That is, a sample injected through the sample injection port 125 may be injected into any one of the plurality of injection holes 112.

The piston 130 may have a cylindrical shape. The piston 130 may include a groove 131 and a power transmission groove 133.

The groove 131 may be formed at a side surface of the piston 130. The groove 131 may be recessed from the side surface of the piston 130.

The power transmission groove 133 may be formed at the upper surface of the piston 130. The power transmission groove 133 may be recessed from the upper surface of the piston 130. The power transmission groove 133 may be coupled to the power unit 20. Power from the power unit 20 is transmitted to the power transmission groove 133, and the piston 130 can be rotated and moved vertically by the power. The piston 130 may be rotated about a rotary shaft 135.

The packing member 140 may have a ring shape. An inner circumferential surface of the packing member 140 may have the same size as the outer circumferential surface of the piston 130. The packing member 140 may be formed of a rubber or silicon material to prevent leakage of the pneumatic pressure and the material.

A plurality of reagent channels 151, an auxiliary channel 152, a first exhaust hole 156 and a second exhaust hole 157 may be formed in the lower housing 150. The lower housing 150 may include a filter unit 153.

The filter unit 153 may have a circular shape. The filter unit 153 may have a shape corresponding to the outer circumferential surface of the piston 130. The piston 130 may be inserted into the filter unit 153.

The plurality of reagent channels 151 may be formed at a region in communication with the filter unit 153. The plurality of reagent channels 151 may be formed to come in contact with the outer circumferential surface of the filter unit 153. The plurality of reagent channels 151 may be connected to the filter unit 153.

The auxiliary channel 152 may be formed to come in contact with the outer circumferential surface of the filter unit 153.

The first exhaust hole 156 and the second exhaust hole 157 may be formed from the upper surface of the lower housing 150 toward the lower surface of the lower housing 150. The plurality of reagent channels 151, the auxiliary channel 152, the first exhaust hole 156 and the second exhaust hole 157 may be formed at an upper section of the lower housing 150.

A valve power transmission hole 158 may be formed in the side surface of the lower housing 150. A partial configuration of the valve power unit 23 may be inserted into the valve power transmission hole 158 to provide power to the valve of the cartridge 100.

FIG. 4 is a plan view showing the chamber housing according to the embodiment, and FIG. 5 is a cross-sectional view showing the chamber housing according to the embodiment.

Referring to FIGS. 4 and 5, the chamber housing 110 according to the embodiment may include a plurality of reagent chambers 114.

The plurality of reagent chambers 114 may be formed to surround the chamber through-hole 113. The plurality of reagent chambers 114 may have spaces divided by a plurality of partition walls 116. The plurality of partition walls 116 may be formed from the outer surface of the chamber housing 110 toward the chamber through-hole 113. The plurality of partition walls 116 can prevent movement of the reagent between the neighboring reagent chambers 114.

A plurality of reagent moving holes 115 may be formed between the plurality of reagent chambers 114 and the chamber through-hole 113. The reagent moving holes 115 may be connected to each of the reagent chambers 114. The reagent accommodated in the reagent chambers 114 can move to the reagent moving holes 115.

Lower sections of the plurality of reagent chambers 114 may have inclinations toward the chamber through-hole 113. The reagent chamber 114 has a cross-sectional area reduced toward the lower region. The reagent moving hole 115 may be formed to be connected to the lower region of the reagent chamber 114. The reagent moving hole 115 may be formed to be connected to the chamber cover 111. That is, the reagent moving hole 115 may be a groove formed at an end of the chamber cover 111.

The plurality of reagent chambers 114 may accommodate reagents. The reagents accommodated in the plurality of reagent chambers 114 may be a plurality of samples for pre-processing the samples. The plurality of reagents may include a reagent for extracting nucleic acid from the sample, a cleaning reagent (washing buffer) for cleaning the extracted nucleic acid, an elution reagent (elution buffer) for eluting the extracted nucleic acid, and a PCR reagent for performing a PCR reaction.

The plurality of reagents may be stored in a liquid phase in the reagent chamber 114, and the reagent in a solid phase and the reagent for dissolving the reagent in the solid phase may be stored in different reagent chambers among the reagent chambers 114. The reagent in the solid phase and the reagent for dissolving the reagent in the solid phase may be stored in the reagent chambers 114 that neighbor each other. The reagent in the solid phase may be a freeze-dried reagent.

The reagent for extracting the nucleic acid from the sample may be a PK reagent (proteinase K), a dissolution buffer (lysis buffer) and ethanol (ETOH). The PK reagent may be used to digest a protein of the sample, and remove a contaminant from the nucleic acid. The PK reagent may be generated by the freeze-dried PK material and PK buffer in the cartridge 100. The PK material and the PK buffer may be stored in different reagent chambers among the reagent chambers 114. The PK material and the PK buffer may be stored in the reagent chambers 114 that neighbor each other.

The dissolution buffer may use various materials capable of dissolving the sample. For example, the dissolution buffer may include a chaotropic agent such as a guanidinium salt (for example, guanidinium thio cyanate), a chelating agent such as ethylenediaminetetraacetic acid (EDTA), and a buffer salt such as trihidroxymethylaminomethane (Tris-HCl). In addition, a non-ionic surfactant may be included. Either a polyethyleneglycol type non-ionic surfactant or a polyhydric alcohol type non-ionic surfactant may be used, preferably, Triton X-100, tween (additives of ethylene oxide of solbitan ester) or 2-mercapto ethanol may be used, or most preferably, Triton X-100 may be used. The dissolution buffer may have non-acidity, for example, neutrality or alkalinity. However, the present invention is not limited thereto and the dissolution buffer may have various materials.

The ethanol may be used to separate the nucleic acid from the sample.

The cleaning reagent may function to clean impurities that may be included with the nucleic acid or the reaction solution used in the previous process to increase purity of the nucleic acid. The cleaning reagent may include ethanol, isopropanol, and so on.

As the elution reagent, various materials that can dissolve the captured nucleic acid may be used. The elution reagent may include water, TE buffer (Tris-Cl, EDTA), or the like.

The PCR reagent may be a material for performing a PCR process, and the PCR reagent may include a primer, DNA polymerase, deoxyribonucleoside triphosphate (hereinafter referred to as "dNTP") for forming new DNA, and so on.

The reagents accommodated in the neighboring reagent chambers 114 of the plurality of reagent chambers 114 may be mixed through the groove 131 of the piston 130.

The plurality of reagent chambers 114 may include first to eighth reagent chambers 114a to 114h. Each of the first to eighth reagent chambers 114a to 114h may accommodate a reagent. The first to eighth reagent chamber 114a to 114h may be connected to first to eighth reagent moving holes 115a to 115h, respectively.

The PK material in the solid phase may be accommodated in the first reagent chamber 114a. The PK buffer in the liquid phase may be accommodated in the second reagent chamber 114b. A dissolution buffer may be accommodated in the third reagent chamber 114c. The ethanol may be accommodated in the fourth reagent chamber 114d. The cleaning reagent may be accommodated in the fifth reagent chamber 114e. The elution reagent may be accommodated in the sixth reagent chamber 114f. The PCR sample may be accommodated in the seventh reagent chamber 114g. The deionized water may be accommodated in the eighth reagent chamber 114h. While the cleaning reagent is shown as being accommodated in one reagent chamber, the cleaning reagent may be accommodated in a plurality of the reagent chambers.

As shown in FIG. 6, when the piston 130 is lowered in the chamber through-hole 113 and the groove 131 is disposed at the reagent moving hole 115 in a horizontal direction, the reagent of the reagent chamber 114 can move into the groove 131 through the reagent moving hole 115.

The groove 131 may be formed to have a width at which it can be formed across the reagent moving holes 115 that neighbor each other. That is, the groove 131 may have a width corresponding to a sum of widths of two of the reagent moving holes 115 and an interval between the neighboring reagent moving holes 115. The groove 131 can provide a first path through which the reagent moves between the neighboring reagent moving holes 115.

For example, when two of the reagent chambers 114 of FIG. 4 that neighbor each other are defined as the first reagent chamber 114a and the second reagent chamber 114b, the reagent moving hole 115 in communication with the first reagent chamber 114a is defined as the first reagent moving hole 115a, and the reagent moving hole 115 in communication with the second reagent chamber 114b is defined as the second reagent moving hole 115b, the groove 131 can be disposed to connect the first reagent moving hole 115a and the second reagent moving hole 115b.

The materials of the first reagent chamber 114a and the second reagent chamber 114b can be mixed by the first reagent moving hole 115a, the second reagent moving hole 115b and the groove 131. The pneumatic unit 30 can completely mix the materials of the first reagent chamber 114a and the second reagent chamber 114b by injecting the pneumatic pressure into the first reagent chamber 114a and the second reagent chamber 114b through the pneumatic pressure injection port 121. For example, when the reagent in the solid phase is accommodated in the first reagent chamber 114a and the reagent for dissolving the reagent in the solid phase is accommodated in the second reagent chamber 114b, the pneumatic unit 30 can dissolve the reagent in the solid phase by alternately injecting the pneumatic pressure into the first reagent chamber 114a and the second reagent chamber 114b.

As the piston 130 is rotated, the neighboring reagent chambers 114 to be mixed can be selected from the plurality of reagent chambers 114. That is, as the piston 130 is angularly moved, the groove 131 can be disposed at the specific neighboring reagent moving holes 115 such that the materials in the neighboring reagent chambers 114 can be selectively mixed. Rotation of the piston 130 may be controlled by the piston power unit 21.

FIG. 7 is a cross-sectional view showing a lower housing according to the embodiment.

Referring to FIG. 7, the plurality of reagent channels 151, the auxiliary channel 152, the filter unit 153, the first exhaust hole 156, the second exhaust hole 157, a valve 160, a discard chamber 171 and a PCR chamber 173 may be formed at the lower housing 150 according to the embodiment.

The plurality of reagent channels 151, the auxiliary channel 152, the first exhaust hole 156 and the second exhaust hole 157 may be formed from the upper surface of the lower housing 150 toward the lower surface.

The plurality of reagent channels 151 may be formed to come in communication with the filter unit 153. The plurality of reagent channels 151 may be formed at a position perpendicular to at least one reagent chamber 114 of the plurality of reagent chambers 114. The reagent channel 151 may be used as a second path configured to move the reagent from the reagent chamber 114 to the filter unit 153.

The auxiliary channel 152 may be formed to come in communication with the filter unit 153. The auxiliary channel 152 may be formed in a position in which it is perpendicular to at least one reagent chamber 114 of the plurality of reagent chambers 114. One end of the auxiliary channel 152 may be connected to the filter unit 153, and the other end of the auxiliary channel 152 may be connected to the PCR chamber 173. The auxiliary channel 152 may be used as a third path configured to move the reagent from the reagent chamber 114 to the PCR chamber 173. The auxiliary channel 152 may be connected to the reagent chamber 114 in which the PCR reagent is accommodated. The auxiliary channel 152 may be formed at a position corresponding to at least one reagent chamber of the seventh reagent chamber 114g and the eighth reagent chamber 114h.

The filter unit 153 may include a membrane filter 154. The membrane filter 154 may be disposed at a lower region of the filter unit 153. The membrane filter 154 can filter the sample and reagent injected from the reagent channel 151. The membrane filter 154 can filter the nucleic acid from the sample and reagent injected from the reagent channel 151. That is, when the sample and the reagent are injected into the membrane filter 154, only the nucleic acid serving as a target material remains at an upper section of the membrane filter 154, and other materials pass through the membrane filter.

A connecting channel 155 may be formed at a lower section of the filter unit 153. The connecting channel 155 may include a first connecting channel 155a, a second connecting channel 155b and a third connecting channel 155c.

The first connecting channel 155a may connect the filter unit 153 and the valve 160, the second connecting channel 155b may connect the valve 160 and the discard chamber 171, and the third connecting channel 155c may connect the valve 160 and the PCR chamber 173.

The valve 160 can move the material in the filter unit 153 into the discard chamber 171 or the PCR chamber 173. The valve 160 can selectively supply the material in the filter unit 153 into the discard chamber 171 or PCR chamber.

The valve 160 may include a valve channel 161. The valve channel 161 may be formed in a V shape. The valve channel 161 may include a first valve channel 161a and a second valve channel 161b. An angle between the first valve channel 161a and the second valve channel 161b may be 120 degrees.

FIG. 8 is a view showing a connecting state of the valve channel according to the embodiment.

The valve 160 may be rotated. The valve 160 may be rotated by the valve power unit 23. The valve 160 may be rotated about a region at which the first valve channel 161a meets the second valve channel 161b.

The valve 160 may be rotated into a first connecting state as shown in FIG. 8a. When the valve 160 is in the first connecting state, the filter unit 153 may be connected to the discard chamber 171. The first connecting channel 155a may be connected to the first valve channel 161a, and the third connecting channel 155c may be connected to the second valve channel 161b.

The filter unit 153 may be connected to the discard chamber 171 through the first connecting channel 155a, the first valve channel 161a, the third connecting channel 155c and the second valve channel 161b. That is, the material accommodated in the filter unit 153 can be transmitted to the discard chamber 171 in the first connecting state.

The valve 160 may be rotated into a second connecting state as shown in FIG. 8b. When the valve 160 is in the second connecting state, the filter unit 153 may be connected to the PCR chamber 173. The first connecting channel 155a may be connected to the second valve channel 161b, and the second connecting channel 155b may be connected to the first valve channel 161a.

The filter unit 153 may be connected to the PCR chamber 173 through the first connecting channel 155a, the second valve channel 161b, the first valve channel 161a and the second connecting channel 155b. That is, the material accommodated in the filter unit 153 can be transmitted to the PCR chamber 173 in the second connecting state.

Returning to FIG. 7, the discard chamber 171 may be connected to the first exhaust hole 156. The discard chamber 171 may be connected to the outside through the first exhaust hole 156. As the discard chamber 171 is connected to the first exhaust hole 156, the air can be discharged to the outside through the first exhaust hole 156 to an extent of a volume of the material injected into the discard chamber 171.

Alternatively, the first exhaust hole 156 may be connected to the pneumatic unit 30 and the pneumatic unit 30 can absorb the gas in the discard chamber 171. As the pneumatic unit 30 absorbs the gas in the discard chamber 171, the material accommodated in the filter unit 153 can be controlled to move into the discard chamber 171.

Alternatively, the first exhaust hole 156 may be used as a passage through which the material accommodated in the discard chamber 171 is discharged.

The PCR chamber 173 may be connected to the second exhaust hole 157. The PCR chamber 173 may be connected to the outside through the second exhaust hole 157. As the PCR chamber 173 is connected to the second exhaust hole 157, the air corresponding to the volume of the material injected into the PCR chamber 173 can be discharged to the outside through the second exhaust hole 157.

Alternatively, the second exhaust hole 157 may be connected to the pneumatic unit 30, and the pneumatic unit 30 can absorb the gas in the PCR chamber 173. As the pneumatic unit 30 absorbs the gas in the PCR chamber 173, the material accommodated in the filter unit 153 can be controlled to move into the PCR chamber 173.

The PCR chamber 173 may have a transparent region formed in at least one surface thereof. As the least one surface of the PCR chamber 173 has the transparent region, the light from the measurement unit 50 can be radiated to the nucleic acid in the PCR chamber 173. The light discharged through the nucleic acid can be output to the measurement unit 50 through the transparent region, and the measurement unit 50 can determine whether the specific nucleic acid is present by analyzing the light.

The PCR chamber 173 may have a reflection region formed in at least one surface thereof. As the reflection region is formed at the PCR chamber 173, the light injected from the measurement unit 50 can be reflected for the purpose of radiating it to the nucleic acid, thereby improving measurement efficiency.

The outer surface of the PCR chamber 173 may be formed of a material having large thermal conductivity. As the PCR chamber 173 is formed of the material having the large thermal conductivity, the heat applied by the temperature adjustment unit 40 can be easily transmitted into the PCR chamber 173.

At least one surface of the PCR chamber 173 may be formed of a material having large thermal conductivity and good reflectance such as aluminum. Accordingly, measurement efficiency and thermal efficiency can be simultaneously increased.

As another example, the lower housing 150 may have a separate PCR region 175. When the PCR region 175 is separately provided, the transparent region or the reflection region may be formed at the PCR region 175.

The PCR region 175 may be connected to the PCR chamber 173. The material accommodated in the PCR chamber 173 may move into the PCR region 175 to perform the PCR reaction and measurement process.

Here, the second exhaust hole 157 may be connected to the PCR region 175. The second exhaust hole 157 can suction the air in the PCR chamber 173 and the PCR region 175 such that the material accommodated in the PCR chamber 173 can be easily moved to the PCR region 175.

FIG. 9 is a view showing a method of controlling an automatic analysis apparatus according to an embodiment of the present invention.

Referring to FIG. 9, the method of controlling the automatic analysis apparatus according to the embodiment includes preparing a cartridge (S210), injecting a sample into the cartridge (S220), generating the PK reagent (S230), extracting nucleic acid from the sample (S240), cleaning the extracted nucleic acid (S250), eluting the nucleic acid (S260), generating a PCR reagent (S270), injecting the PCR reagent (S280), and performing amplification and measurement (S290).

Hereinafter, the method of controlling the automatic analysis apparatus will be described with reference to the accompanying drawings.

In preparing the cartridge (S210), the reagent is injected into the plurality of reagent chambers 114 of the cartridge 100. The cartridge 100 may be provided in a state in which the reagent is injected into the plurality of reagent chambers 114. The plurality of reagent chambers 114 may include the first to eighth reagent chamber 114a to 114h as shown in FIG. 4. The reagents may be accommodated in each of the first to eighth reagent chambers 114a to 114h.

The PK material in the solid phase may be accommodated in the first reagent chamber 114a. The PK buffer in the liquid phase may be accommodated in the second reagent chamber 114b. The dissolution buffer may be accommodated in the third reagent chamber 114c. The ethanol may be accommodated in the fourth reagent chamber 114d. The cleaning reagent may be accommodated in the fifth reagent chamber 114e. The elution reagent may be accommodated in the sixth reagent chamber 114f. The PCR sample may be accommodated in the seventh reagent chamber 114g. The deionized water may be accommodated in the eighth reagent chamber 114h. While the cleaning reagent is shown as being accommodated in one reagent chamber, the cleaning reagent may be accommodated in a plurality of the reagent chambers.

A sample 101 is injected into the prepared cartridge 100 as shown in FIG. 10 (S220).

The sample may be injected into any one reagent chamber of the plurality of reagent chambers 114. The sample may be injected into the reagent chamber 114, in which the dissolution buffer (lysis buffer) is accommodated, in the plurality of reagent chambers 114. The sample may be injected into the third reagent chamber 114c. The sample may be blood.

When the sample 101 is injected, the cartridge 100 generates the PK reagent (S230).

Referring to FIG. 11, the PK material in the solid phase and the PK buffer in the liquid phase are accommodated in the first reagent chamber 114a and the second reagent chamber 114b, which are adjacent to each other, respectively.

The piston 130 is lowered and the groove 131 is disposed at the first reagent moving hole 115a and the second reagent moving hole 115b at a horizontal region. A first path is formed by the groove 131, the first reagent moving hole 115a and the second reagent moving hole 115b, and the first reagent chamber 114a and the second reagent chamber 114b come in communication with each other through the first path.

Here, the pneumatic pressure is alternately supplied into the first reagent chamber 114a and the second reagent chamber 114b by the pneumatic unit 30, and the PK material and the PK buffer are mixed to generate the PK reagent. The pneumatic unit 30 alternately supplies the pneumatic pressure into the first reagent chamber 114a and the second reagent chamber 114b to generate the PK reagent, and when the PK reagent is generated, supplies the pneumatic pressure into the first reagent chamber 114a to inject the PK reagent into the second reagent chamber 114b.

Next, the cartridge 100 performs a nucleic acid extraction process (S240).

First, the PK reagent injected into the second reagent chamber 114b is mixed with the dissolution buffer (lysis buffer) and the sample accommodated in the third reagent chamber 114c.

As shown in FIG. 12, the piston 130 is rotated and the groove 131 is moved to a position corresponding to the second reagent chamber 114b and the third reagent chamber 114c. The groove 131 is disposed at the second reagent moving hole 115b and the third reagent moving hole 115c at a horizontal region. The first path is formed by the groove 131, the second reagent moving hole 115b and the third reagent moving hole 115c, and the second reagent chamber 114b and the third reagent chamber 114c come in communication with each other through the first path.

Here, the pneumatic pressure is alternately supplied into the second reagent chamber 114b and the third reagent chamber 114c by the pneumatic unit 30 to mix the PK reagent with the dissolution buffer and the sample. After the PK reagent is mixed with the dissolution buffer and the sample, the pneumatic pressure is supplied into the second reagent chamber 114b, and the material obtained by mixing the PK reagent with the dissolution buffer and the sample is injected into the third reagent chamber 114c.

Next, the material obtained by mixing the PK reagent with the dissolution buffer and the sample is mixed with the ethanol (ETOH) accommodated in the fourth reagent chamber 114d.

As shown in FIG. 13, the piston 130 is rotated and the groove 131 is moved to a position corresponding to the third reagent chamber 114c and the fourth reagent chamber 114d. The groove 131 is disposed at the third reagent moving hole 115c and the fourth the reagent moving hole 115d at a horizontal region. The first path is formed by the groove 131, the third reagent moving hole 115c and the fourth reagent moving hole 115d, and the third reagent chamber 114c and the fourth reagent chamber 114d come in communication with each other through the first path.

Here, the pneumatic pressure is alternately supplied into the third reagent chamber 114c and the fourth the reagent chamber 114d by the pneumatic unit 30, and the material obtained by mixing the PK reagent with the dissolution buffer and the sample is mixed with the ethanol. The material obtained by mixing the PK reagent with the dissolution buffer and the sample is mixed with the ethanol to extract the nucleic acid from the sample.

When the nucleic acid is extracted, as shown in FIG. 14, the piston 130 is lowered and the groove 131 is disposed at a region corresponding to the reagent moving hole 115 and the reagent channel 151. That is, the groove 131 is disposed at a position at which the third reagent moving hole 115c and the fourth reagent moving hole 115d correspond to the reagent channel 151. A second path is formed by the groove 131, the third reagent moving hole 115c, the fourth reagent moving hole 115d and the reagent channel 151, and the extracted nucleic acid and the material accommodated in the third reagent chamber 114c and fourth the reagent chamber 114d are moved into the filter unit 153.

Here, the pneumatic pressure from the pneumatic unit 30 can be injected into the third reagent chamber 114c and the fourth reagent chamber 114d, and the material accommodated in the third reagent chamber 114c and the fourth reagent chamber 114d can be smoothly moved into the filter unit 153.

The valve 160 may be rotated into a first connecting state. The valve 160 connects the filter unit 153) and the discard chamber 171 to move materials other than the extracted nucleic acid into the discard chamber 171. The extracted nucleic acid remains on the membrane filter 154.

Next, a cleaning process of the extracted nucleic acid is performed (S250).

The cleaning reagent accommodated in the fifth reagent chamber 114e is moved into the filter unit 153 and the nucleic acid remaining in the filter unit 153 is cleaned.

As shown in FIG. 15, the piston 130 is rotated and the groove 131 is disposed at a position corresponding to the fifth the reagent chamber 114e. Since the piston 130 is in a lowered state, the second path is formed by the groove 131, the fifth reagent moving hole 115e and the reagent channel 151. Since the cleaning reagent accommodated in the fifth reagent chamber 114e is moved into the filter unit 153 through the second path and the valve 160 maintains the second connecting state, the cleaning reagent passes through the filter unit 153 and moves into the discard chamber 171. The pneumatic pressure from the pneumatic unit 30 can be injected into the fifth reagent chamber 114e in this process as well.

The nucleic acid remaining on the membrane filter 154 can be cleaned by the cleaning reagent.

The cleaning process can be repeatedly performed. When the cleaning process is repeatedly performed, the cleaning reagent is accommodated in the plurality of reagent chambers 114, the piston 130 is angularly rotated to dispose the groove 131 in the reagent chamber 114, and the cleaning reagent of the reagent chamber 114 is sequentially supplied into the filter unit 153 to repeatedly clean the nucleic acid.

After the nucleic acid is cleaned, the elution process is performed (S260).

The elution reagent is injected into the filter unit 153 and the nucleic acid is eluted.

As shown in FIG. 16, the valve 160 is rotated into a second connecting state. The valve 160 can connect the filter unit 153 and the PCR chamber 173.

The piston 130 is rotated and the groove 131 is moved to a position corresponding to the sixth reagent chamber 114f, in which the elution reagent is accommodated, to form the second path. The elution reagent is moved into the filter unit 153 through the second path, and the extracted nucleic acid remaining on the filter unit 153 is eluted to move the nucleic acid into the PCR chamber 173.

After the elution process is performed with respect to the nucleic acid, the PCR reagent is generated (S 270).

As shown in FIGS. 17 and 18, the piston 130 is raised and rotated to dispose the groove 131 at a position corresponding to the seventh reagent chamber 114g and the eighth reagent chamber 114h, in which the PCR sample and the deionized water (DW) are accommodated.

The groove 131 is disposed at the seventh reagent moving hole 115g and the eighth reagent moving hole 115h at a horizontal region. The first path is formed by the groove 131, the seventh reagent moving hole 115g and the eighth reagent moving hole 115h, and the seventh reagent chamber 114g and the eighth reagent chamber 114h come in communication with each other through the first path.

Here, the pneumatic pressure is alternately supplied into the seventh reagent chamber 114g and the eighth reagent chamber 114h by the pneumatic unit 30 to mix the PCR sample with the deionized water to generate the PCR reagent.

When the PCR reagent is generated, the cartridge 100 injects the PCR reagent into the PCR chamber 173 (S280).

Referring to FIG. 19, since the auxiliary channel 152 is formed at a position corresponding to at least one reagent chamber of the seventh reagent chamber 114g and the eighth reagent chamber 114h, the PCR reagent is injected into the PCR chamber 173 by the auxiliary channel 152.

The piston 130 is lowered to form a third path between at least one reagent chamber of the seventh reagent chamber 114g and the eighth reagent chamber 114h, and the auxiliary channel 152. That is, the third path is formed by at least one reagent chamber of the seventh reagent chamber 114g and the eighth reagent chamber 114h, the groove 131, and the auxiliary channel 152.

The nucleic acid eluted through the third path and the PCR reagent are mixed in the PCR chamber 173. As the PCR reagent is moved into the PCR chamber 173 through a separate third path, contamination of the PCR reagent can be prevented to obtain an accurate nucleic acid analysis result.

When the PCR reagent is mixed with the eluted nucleic acid, PCR with respect to the nucleic acid accommodated in the PCR chamber 173 is performed. The PCR chamber 173 is repeatedly heated and cooled by the temperature adjustment unit 40 to amplify the nucleic acid, and in the process of amplifying the nucleic acid, the measurement unit 50 can measure whether the specific nucleic acid is present to detect a pathogenic organism or the like.

The processes of performing extraction, amplification, and measurement of the nucleic acid using one apparatus can be simultaneously performed to reduce a measurement time, prevent contamination, and obtain an accurate measurement value.

It will be apparent to those skilled in the art that various modifications can be made to the above-described exemplary embodiments of the present invention without departing from the spirit or scope of the invention. Thus, it is intended that the present invention cover all such modifications provided they come within the scope of the appended claims and their equivalents.

According to the present invention, the present invention use for reducing a processing time by mixing a reagent using a channel of a piston, transmitting the mixed reagent to a filter unit, and extracting and cleaning nucleic acid.

According to the present invention, the present invention use for prevent contamination of a PCR reagent by moving the PCR reagent using a separate auxiliary channel.

1: automatic analysis apparatus

10: control unit

20: power unit

30: pneumatic unit

40: temperature adjustment unit

50: measurement unit

100: cartridge

110: chamber housing

111: chamber cover

112: injection hole

113: chamber through-hole

114: reagent chamber

115: reagent moving hole

120: upper cover

121: pneumatic pressure injection port

130: piston

131: groove

133: power transmission groove

135: rotary shaft

140: packing member

150: lower housing

151: reagent channel

152: auxiliary channel

153: filter unit

155: connecting channel

156: first exhaust hole

157: second exhaust hole

160: valve

161: valve channel

171: discard chamber

173: PCR chamber

Claims

A cartridge comprising:

a plurality of reagent chambers;

a discard chamber;

a PCR chamber; and

a piston provided with a channel in which a first path and a second path are provided,

wherein the first path is used to move a reagent between the plurality of reagent chambers, and

the second path is used to move a material in the reagent chamber into the discard chamber or the PCR chamber.
The cartridge of claim 1, wherein the channel is a groove formed in the piston.
The cartridge of claim 1, wherein the first path is provided by rotation of the piston.
The cartridge of claim 1, wherein the second path is provided by vertical movement of the piston.
The cartridge of claim 3, wherein the first path is a path through which a material is moved between the neighboring reagent chambers.
The cartridge of claim 1, further comprising a valve configured to selectively supply the material from the second path into the discard chamber or the PCR chamber.
The cartridge of claim 6, further comprising a filter unit disposed between the second path and the valve and configured to filter a target material from the material from the second path.
The cartridge of claim 7, wherein the filter unit comprises a membrane filter.
The cartridge of claim 1, further comprising an auxiliary channel configured to connect at least one of the plurality of reagent chambers and the PCR chamber.
The cartridge of claim 9, wherein the auxiliary channel is formed at a lower housing.
The cartridge of claim 9, wherein the auxiliary channel is connected to at least one of the plurality of reagent chambers through the channel.
The cartridge of claim 1, wherein a partial region of an outer surface of the PCR chamber is a light transmission region.
The cartridge of claim 1, wherein a pneumatic pressure is supplied into the plurality of reagent chambers.
The cartridge of claim 7, further comprising a packing member disposed at an outer circumferential surface of a central region of the piston.
The cartridge of claim 14, wherein the central region is an intermediate region between the reagent chamber and the filter unit.
The cartridge of claim 6, wherein the valve comprises a V-shaped channel.