KR101437261B1 - Sample treatment chip for rotary pcr processing, rotary pcr apparatus with the same and rotary pcr method using the same - Google Patents

Abstract

According to the present invention, there is provided a unit sample processing apparatus comprising a unit sample processing unit spaced apart from a rotation center in a radial direction, the unit sample processing unit including a trapping passage having an inlet and an outlet, part; A sample chamber located radially inward of the target substance capturing portion and connected to an inlet of the trapping passage and providing a space in which a sample is stored; A washing buffer liquid chamber located radially inward of the target substance capturing portion and providing a space in which the washing buffer solution is stored; A wash buffer liquid passage located between the wash buffer solution chamber and the inlet of the trap passage; An elution buffer solution chamber located radially inward of the target substance capturing part and providing a space in which an elution buffer solution is stored; And an eluting buffer solution introducing passage positioned between the eluting buffer solution chamber and an inlet of the trapping passage, the washing buffer solution introducing passage including an inner switching curve portion for switching the flow of the washing buffer solution from the radially inner to the outer Wherein the eluting buffer solution introducing passage includes a first inner switching curve portion and a second inner switching curve portion which are sequentially arranged in the flow direction and which convert the flow of the eluting buffer solution from the radially inner side to the outer side, A sample processing chip for a rotary PCR process is provided.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a sample processing chip for a rotary PCR process, a rotary PCR apparatus having the same, and a rotary PCR method using the same. BACKGROUND ART [0002]

The present invention relates to PCR, and more particularly, to a sample processing chip for performing a PCR process by a rotation method, a PCR apparatus having the same, and a PCR method using the same.

Generally, DNA amplification technology has been widely used for research and development and diagnosis purposes in life sciences, genetic engineering and medical fields, and in particular, DNA amplification technology by polymerase chain reaction (PCR) has been widely used . The polymerase chain reaction (PCR) is used to amplify a specific DNA sequence in the genome as necessary.

The PCR process involves three steps: denaturation, annealing, and elongation. In the first stage of denaturation, the two strands of DNA are heated and separated. Each separated DNA serves as a template. In the second step, the binding step, the primers bind to the template DNA. The temperature in the binding step is an important factor in determining the accuracy of the reaction. If the temperature is too high, the primer will bind too weakly to the template DNA, resulting in very little amplified DNA product. If the temperature is too low, undesirable DNA can be amplified because the primer binds nonspecifically. In the third step, the kidney step, heat-resistant DNA polymerase makes new DNA from template DNA.

In general, a substance such as DNA or RNA (hereinafter referred to as a "target substance") which is subjected to a PCR process is separated and purified by pretreatment of a sample containing the target substance. Generally, pretreatment of a sample is carried out by allowing the sample to flow to a trap means such as silica beads to adsorb the target substance to the trap means and flow the wash buffer solution as a trap means to remove the component other than the target substance from the trap means, And separating the target material adsorbed to the capturing means by flowing into the capturing means.

An object of the present invention is to provide a sample processing chip for a rotary PCR process capable of performing a PCR process including a pretreatment of a sample in a rotating manner, a rotation PCR device having the same, and a rotation PCR method using the same.

According to an aspect of the present invention,

And a unit sample processing unit disposed radially from the rotation center, wherein the unit sample processing unit includes a target substance capturing unit including a capturing channel having an inlet and an outlet and a capturing means filled in the capturing channel; A sample chamber located radially inward of the target substance capturing portion and connected to an inlet of the trapping passage and providing a space in which a sample is stored; A washing buffer liquid chamber located radially inward of the target substance capturing portion and providing a space in which the washing buffer solution is stored; A wash buffer liquid passage located between the wash buffer solution chamber and the inlet of the trap passage; An elution buffer solution chamber located radially inward of the target substance capturing part and providing a space in which an elution buffer solution is stored; And an eluting buffer solution introducing passage positioned between the eluting buffer solution chamber and an inlet of the trapping passage, the washing buffer solution introducing passage including an inner switching curve portion for switching the flow of the washing buffer solution from the radially inner to the outer Wherein the eluting buffer solution introducing passage includes a first inner switching curve portion and a second inner switching curve portion which are sequentially arranged in the flow direction and which convert the flow of the eluting buffer solution from the radially inner side to the outer side, A sample processing chip for a rotary PCR process is provided.

The washing buffer solution introducing passage may further include an outer switching curve portion located upstream of the inner switching curve portion and switching the flow of the washing buffer solution from the radially outer side to the inner side.

Wherein the eluting buffer solution introducing passage includes a first outside switching curve portion located on the upstream side of the first inside switching curve portion and switching the flow of the eluting buffer solution from the outside to the inside in the radial direction, And a second outer switching curve portion for switching the flow of the eluting buffer solution from the radially outer side to the inner side.

The trapping passage connects the inlet and the outlet in a zigzag shape, and the inlet and the outlet may be located radially inward and outward, respectively.

The unit sample processing unit may further include a discharge passage located radially outward of the target substance capturing unit and connected to the outlet of the trapping passage.

The unit sample processing unit may further include a circulation passage for connecting the discharge passage to the sample chamber, the wash buffer solution chamber, and the elution buffer solution chamber.

A plurality of unit sample processing units may be disposed along the circumferential direction.

The trapping means may be a silica-based material.

The target material trapping portion may have a weir structure.

According to another aspect of the present invention, there is provided a sample processing chip for the rotary PCR process, And a rotation driving unit for rotating the sample processing chip.

The rotary PCR apparatus may further include a temperature regulator in which the sample processing chip is accommodated and a heating region for providing a required temperature for each step of the PCR process is disposed along the circumferential direction.

According to another aspect of the present invention,

As the rotation PCR method using the sample processing chip for the rotary PCR process,

Preparing a sample processing chip in which a sample is injected into the sample chamber, a washing buffer solution is injected into the washing buffer solution chamber, and an elution buffer solution is injected into the elution buffer solution chamber; And a preprocessing step of rotating the sample processing chip to perform a pretreatment on the sample, wherein the preprocessing step is a step of rotating the sample processing chip at a first rotation speed to move the target Material capture stage; Wherein the washing buffer solution is passed through the inside switching curve portion of the washing buffer solution introducing passage so that the eluting buffer solution is passed through the elution buffer solution A first introducing step of passing the first inner switching curve portion of the introduction passage; A washing buffer solution loading step of increasing the rotational speed of the sample processing chip to a third rotational speed that is faster than the second rotational speed to introduce the washing buffer solution into the target substance capturing part; A second introduction step of decelerating the rotation speed of the sample processing chip at a fourth rotation speed slower than the third rotation speed so that the elution buffer solution passes through the second inside transition curve portion of the elution buffer solution introduction passage; And an elution buffer solution loading step of increasing the rotation speed of the sample processing chip to a fifth rotation speed faster than the fourth rotation speed so as to introduce the elution buffer solution into the target substance capturing part.

The second rotation speed and the fourth rotation speed may be the same.

The rotational PCR method may further include a target material collecting step of collecting target material by increasing the rotational speed of the sample processing chip to a sixth rotational speed that is higher than the fifth rotational speed after the elution buffer solution loading step.

According to the present invention, all of the objects of the present invention described above can be achieved. Specifically, since the process of introducing the washing buffer solution and the elution buffer solution into the target substance capturing part is controlled by controlling the rotation speed, the sample pretreatment can be easily performed without forming the microvalve.

1 is a perspective view of a rotating PCR device according to an embodiment of the present invention.
2 is a perspective view of the sample processing chip for the rotary PCR process shown in FIG.
FIG. 3 is a plan view of the unit sample processing unit shown in FIG. 2. FIG.
FIG. 4 is a plan view schematically showing the configuration of the temperature control unit shown in FIG. 1. FIG.
5 is a flowchart showing a rotation PCR method using the rotation PCR apparatus shown in FIG.
FIG. 6 is a flowchart showing a detailed process of the preprocessing step shown in FIG.
FIGS. 7A to 7G are diagrams illustrating a process in which the preprocessing step shown in FIG. 6 is performed in the unit sample processing unit shown in FIG.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings.

1 is a perspective view of a rotating PCR device according to an embodiment of the present invention. Referring to FIG. 1, the rotating PCR apparatus 100 includes a sample processing chip 110, a temperature controller 180, and a chip rotation driver 190. The rotary PCR apparatus 100 performs a PCR process including pretreatment of a sample while rotating the sample processing chip 110.

Fig. 2 is a perspective view of the sample processing chip 110 for the rotary PCR process shown in Fig. Referring to FIGS. 1 and 2, a sample processing chip 110 for a rotary PCR process is generally disk-shaped and rotates about a rotation axis X passing through the center by a rotation drive unit 190. The sample processing chip 110 for the rotation PCR process has a plurality of unit sample processing units 120 arranged in order along the circumferential direction. In the present embodiment, the unit sample processing unit is described as having four unit samples, but the present invention is not limited thereto. The present invention also includes the case where the sample processing chip 110 for a rotary PCR process has one to three unit sample processing units or five or more unit sample processing units. Since all four unit sample processing units 120 have the same configuration, only one unit sample processing unit 120 will be described in detail here. The unit sample processing unit 120 is formed inside the sample processing chip 110 for the rotation PCR process. For example, a sample processing chip 110 for a rotating PCR process is manufactured by processing a disk-shaped polycarbonate (PC) having a thickness of approximately 1 mm using a CNC milling machine to form a pattern of a unit sample processing unit 120 And then adhering a PC film having a thickness of approximately 100 mu m to the processed surface.

FIG. 3 is a plan view of the unit sample processing unit 120 shown in FIG. 2 and 3, the unit sample processing unit 120 includes a target substance capturing unit 125, a sample chamber 130, a sample introduction channel 135, a washing buffer chamber 140 A washing buffer solution introducing passage 145, an elution buffer chamber 150, an eluting buffer solution introducing passage 155, a connecting passage 160a, a discharging passage 160, A waste material chamber 165, a target material chamber 170, and a circulation passage 175. [

The target material capturing portion 125 has a trapping passage 126 in a zigzag shape and trapping means 128 such as a silica bead filled in the trapping passage 126. In the target substance capturing part 125, the substance containing the target substance in the sample introduced into the trapping passage 126 is captured by the capturing means 128. An inlet 126a and an outlet 126b are located at both ends of the trapping passage 126. [ The inlet 126a is positioned radially inward with respect to the axis of rotation X and the outlet 126b is located radially outward. In the capturing means 128, a substance containing the target substance is adsorbed from the sample. In this embodiment, the capturing means 128 is described as being a silica bead. Although not shown in detail, the trapping passage 126 has a weir structure on the side of the outlet 126b, so that the trapping means is constructed inside.

The sample chamber 130 is positioned radially inward of the inlet 126a of the trapping passage 126. [ A sample is stored in the sample chamber 130. In the sample processing chip 110, a sample injection hole 110a for injecting a sample into the sample chamber 130 is formed.

The sample introduction channel 135 connects the sample chamber 130 and the inlet 126a of the capture channel 126. [ The sample stored in the sample chamber 130 flows into the trapping passage 126 through the sample introduction passage 135 by the centrifugal force generated when the sample processing chip 110 rotates.

The washing buffer chamber 140 is positioned radially inward of the inlet 126a of the trapping passage 126 and positioned closer to the axis of rotation X than the sample chamber 130. [ The wash buffer solution chamber 140 stores the wash buffer solution. The wash buffer solution rinses and removes the remaining components from the trapped means 128, except for the target material, from the trapped means 128. The sample processing chip 110 is provided with a washing buffer solution injection hole 110b for injecting the washing buffer solution into the washing buffer solution chamber 140. [ The washing buffer solution stored in the washing buffer solution chamber 140 flows into the washing buffer solution introducing passage 145 by the centrifugal force generated when the sample processing chip 110 rotates.

The washing buffer solution introducing passage 145 extends from the washing buffer solution chamber 140 and is connected to the connecting passage 160a. The washing buffer solution stored in the washing buffer solution chamber 140 through the washing buffer solution introducing passage 145 flows into the trapping passage 126 via the connecting passage 160a.

Referring to FIG. 3, the wash buffer solution introducing passageway 145 extends from the radial end of the wash buffer solution chamber 140, generally radially outward, and then smoothly curves and extends in a radially inward direction Followed by a soft curve and the direction is turned radially outward to extend. Accordingly, the washing buffer solution introducing passage 145 has the outside switching curve portion 146 provided on the upstream side and the inside switching curve portion 147 provided on the downstream side along the flow direction of the washing buffer solution. The outer switching curve portion 146 switches the flow of the washing buffer solution from the radially outer side to the inner side and flows into the inner switching curve portion 147. The inner switching curve portion 147 turns the flow of the washing buffer solution radially outward and flows into the connecting passage 160a.

The elution buffer chamber 150 is positioned radially inward of the inlet 126a of the trapping passage 126 and the distance from the axis of rotation X is similar to the wash buffer solution chamber 140. In the elution buffer solution chamber 150, an elution buffer solution is stored. The elution buffer solution separates the target material adsorbed by the trapping means 128 from the trapping means 128. In the sample processing chip 110, an elution buffer solution injection hole 110c for injecting an elution buffer solution into the elution buffer solution chamber 150 is formed.

The elution buffer solution introduction passage 155 extends from the elution buffer solution chamber 150 and is connected to the connection passage 160a. The elution buffer solution stored in the elution buffer solution chamber 150 through the elution buffer solution introduction passage 155 flows into the trapping passage 126 via the connection passage 160a.

Referring to FIG. 3, the eluting buffer solution introducing passage 155 extends from the radial end of the eluting buffer solution chamber 150 to a generally radially outward direction, And then extends in a radially outward direction while being smoothly curved and extends in a radially outward direction while being smoothly curved and extending in a radially inward direction. Accordingly, the eluting buffer solution introducing passage 155 has the first outside switching curve portion 156, the first inside switching curve portion 157, and the second inside switching curve portion 157, which are arranged in order from the upstream side to the downstream side along the flow direction of the eluent, A second outside switching curve portion 158, and a second inside switching curve portion 159. The second inside switching curve portion 159 and the second inside switching curve portion 159 are the same. The first outer switching curve portion 156 switches the flow of the eluting buffer solution from the radially outer side to the inner side to introduce the eluting buffer solution into the first inner switching curve portion 157. The first inside switching curve portion 157 switches the flow of the eluting buffer solution from the radially inner side to the outer side to introduce the eluting buffer solution into the second outer switching curve portion 158. The second outer switching curve portion 158 switches the flow of the eluting buffer solution from the radially outer side to the inward side to introduce the eluting buffer solution into the second inner switching curve portion 159. The second inside switching curve portion 159 switches the flow of the eluting buffer solution from the inside to the outside of the elution buffer to introduce the eluting buffer solution into the connection passage 160a.

The connecting passage 160a is connected to the ends of the washing buffer solution introducing passage 145 and the eluting buffer solution introducing passage 155 and extends radially outwardly to be connected to the inlet 126a of the catching passage 126. [

The discharge passage 160 extends from the outlet 126b of the trapping passage 126. [ The discharge passage 160 is positioned radially outwardly away from the outlet 126b of the trapping passage 126 along the path so that the liquid discharged from the trapping passage 126 as the sample processing chip 110 rotates And moves along the discharge passage 160. The discharge passage 160 has a connection section 162 extending such that the downstream side thereof is located in the forward rotational direction. The waste liquid chamber 165 and the target material chamber 170 are connected to the connection section 162.

The waste liquid chamber 165 is located radially outward of the connection section 162 of the discharge passage 160. The waste liquid chamber 165 is connected to the connection section 162 of the discharge passage 160 through the first connection passage 166. In the waste liquid chamber 165, unnecessary components other than the target substance are stored.

The target material chamber 170 is located radially outward of the connection section 162 of the discharge passage 160. The target material chamber 170 is connected to the connection section 162 of the discharge passage 160 through the second connection passage 171. The portion of the second connection passage 171 that is connected to the connection section 162 is positioned downstream of the portion of the first connection passage 166 that is connected to the connection section 162.

The circulation passage 175 extends from the downstream end of the discharge passage 160 and is connected to the sample chamber 130, the wash buffer solution chamber 140 and the elution buffer solution chamber 150, respectively. The circulation passage 175 facilitates the movement of each liquid.

Referring to FIG. 1, the temperature regulator 180 includes a lower member 181 and an upper member 182. The temperature regulator 180 regulates the temperature required for the PCR process. On the upper surface of the lower member 181, a sample processing chip 110 is rotatably mounted on the temperature adjusting portion 180. The upper member 182 moves up and down with respect to the lower member 181 to receive the sample processing chip 110 therein. 185b, 185c, 185d, 185e, 185f, 185g, 185h, 185i, 185j, 185k, 185m are formed in order along the circumferential direction in the temperature regulating part 180 do. An insulator or cooling means 186 is provided between the heating regions 185a, 185b, 185c, 185d, 185e, 185f, 185g, 185h, 185i, 185j, 185k, The heating zones 185a, 185b, 185c, 185d, 185e, 185f, 185g, 185h, 185i, 185j, 185k, 185m may be formed by suitable heating means such as heating blocks. A plurality of heating zones 185a, 185b, 185c, 185d, 185e, 185f, 185g, 185h, 185i, 185j, 185k, 185m are arranged along the circumferential direction, The fourth heating zone 185d, the fifth heating zone 185e, the sixth heating zone 185f, the seventh heating zone 185g, the eighth heating zone 185h, the ninth heating zone 185d, A heating zone 185i, a tenth heating zone 185j, an eleventh heating zone 185k, and a twelfth heating zone 185m. The first, fourth, seventh and tenth heating zones 185a, 185d, 185g and 185j provide the temperatures required for the denaturation step in the PCR process. The second, fifth, eighth, and eleventh heating zones 185b, 185e, 185h, and 185k provide the temperatures required for the coupling step in the PCR process. The third, sixth, ninth, and twelfth heating zones 185c, 185f, 185i, and 185m provide the temperatures required for the extension step in the PCR process. 185h and 185i (185j, 185h and 185i), 185j, 185k and 185m constitute a unit temperature control section. That is, the temperature regulating unit 180 has four unit temperature regulating sections, each of which corresponds to each of the four unit sample processing units 120 located along the circumferential direction on the sample processing chip 110, Temperature.

1, the chip rotation driving unit 190 includes a rotation driving motor for rotating the sample processing chip 110 about the rotation axis X. As shown in FIG. The chip rotation driving unit 190 rotates the sample processing chip 110 to generate a centrifugal force necessary for liquid movement in the preprocessing step and controls the temperature of each target material chamber 170 of the sample processing chip 110 (180) so as to be positioned in the required heating area.

An embodiment of the PCR method using the PCR apparatus described with reference to FIG. 1 to FIG. 4 will now be described with reference to FIG. Referring to FIG. 5, the PCR method includes a sample preparation chip preparation step S10, a sample processing chip mounting step S20, a preprocessing step S30, and a PCR execution step S40.

In the sample processing chip preparing step (S10), the sample, the washing buffer solution and the eluting buffer solution are supplied to the sample processing chip 110 as shown in FIG. Specifically, the sample is injected into the sample chamber 130, the washing buffer solution is injected into the storage part 140, and the elution buffer solution is injected into the elution buffer solution chamber 150.

In the sample processing chip mounting step S20, the sample processing chip 110 into which the sample, the washing buffer solution, and the eluting buffer solution are injected is loaded in the temperature controller 180 through the sample preparation chip preparation step S10. The sample processing chip 110 accommodated in the temperature adjusting unit 180 is connected to the rotation driving unit 190 and is rotatable.

In the preprocessing step S30, the target material contained in the sample stored in the sample chamber 130 of the sample processing chip 110 is separated from other unnecessary components and stored in the target material chamber 170. A more detailed process of the preprocessing step S30 is shown as a flowchart in Fig. The preprocessing step S30 includes the steps of capturing the target material S31, the first introduction step S32, the washing liquid loading step S33, the second introduction step S34, the eluting buffer solution loading step S35, , And a target material collecting step (S36). 7A to 7G show the state of the unit sample processing unit 120 in each step of the preprocessing step S30.

The target material capturing step S31 is performed by rotating the sample processing chip 110 around a rotation axis X for a predetermined time (for example, 4 minutes) at a first rotational speed (for example, 5000 RPM) . In the target material capturing step (S31), the target material contained in the sample is adsorbed to the capturing means (128). 7A and 7B show the states of the unit sample processing unit 120 at the beginning and end of the target substance capturing step S31. Referring to FIG. 7A, the sample S stored in the sample chamber 130 flows into the target material capturing unit 125 by centrifugal force. The substance containing the target substance in the sample S is adsorbed to the capturing means 128 as it passes through the capturing portion 125. [ The entire wash buffer solution W and the entire elution buffer solution E remain in the wash buffer solution chamber 140 and elution buffer solution chamber 150, respectively. Referring to FIG. 7B, the component S1 not captured in the trapping means 128 in the sample by the centrifugal force is stored in the waste liquid chamber 165 via the discharge passage 160. As shown in FIG. The washing buffer solution W is maintained in a state of being moved to one position between the outside switching curve portion 146 and the inside switching curve portion 147 on the washing buffer solution introducing passage 145. The eluting buffer solution E is maintained in a state of being moved to a position between the first outside switching curve portion 156 and the first inside switching curve portion 157 on the eluting buffer solution introducing path 155.

In the first introduction step S32, after the target material capturing step S31, the sample processing chip 110 rotates at a second rotation speed (for example, 100 RPM) substantially lower than the first rotation speed about the rotation axis X, For a predetermined time (for example, 30 seconds). FIG. 7C shows the state of the unit sample processing unit 120 in the first introduction step (S32). Referring to FIG. 7C, the washing buffer solution W flows into the target substance capturing part 125 via the inner switching curve part 147 and the connecting passage 160a. In the washing buffer solution introducing step (S32), the eluting buffer solution (E) passes through the first inside switching curve portion (157), and a portion between the first inside switching curve portion (157) and the second outside switching curve portion And remains in the moved state to the position. The passage of the washing buffer solution W through the inner switching curve portion 147 and the passage of the eluting buffer solution E through the first inside switching curve portion 157 are caused by the centrifugal force reduction due to a sudden deceleration.

The washing buffer solution loading step S33 is a step in which after the first introduction step S32 the sample processing chip 110 is rotated at a third rotational speed which is significantly higher than the second rotational speed about the rotational axis X 5000 RPM, which is equal to the rotation speed) and is rotated for a predetermined time (for example, 30 seconds). In the washing buffer solution loading step S33, the washing buffer solution W passes through the target material capturing part 125 by the centrifugal force, and unnecessary components other than the target material in the target material capturing part 125 Wash and remove. The washing buffer solution W1 passing through the target substance capturing part 125 is stored in the waste liquid chamber 165 as shown in Fig. 7D. At this time, the sample component S1 stored in the waste solution chamber 165 and the wash buffer solution W1 completely fill the waste solution chamber 165. [ 7B, the elution buffer solution E passes through the washing buffer solution loading step S33, and the elution buffer solution E passes through the elution buffer solution introducing passage 155 as shown in FIG. 7D to the second inside switching curve part 158 and the second inside switching curve part (159), as shown in Fig.

In the second introduction step S34, after the washing buffer solution introduction step S33, the sample processing chip 110 rotates about the rotation axis X at a fourth rotation speed (for example, the second rotation speed (100 RPM, which is the same as the rotation speed) and is rotated for a predetermined time (for example, 30 seconds). FIG. 7E shows the state of the unit sample processing unit 120 in the second introduction step (S34). Referring to FIG. 7E, the eluting buffer solution E is moved to the connection passage 160a through the second inside switching curve portion 159. [ The passage of the eluting buffer solution (E) through the second inside switching curve portion (159) is caused by a decrease in centrifugal force due to a sudden deceleration.

The eluting buffer solution loading step S35 is performed after the second introduction step S34 after the sample processing chip 110 has rotated at a fifth rotational speed (for example, 2000 RPM ) And is rotated for a predetermined time (for example, 30 seconds). 7F, in the elution buffer solution loading step S35, the elution buffer solution E passes through the target material capturing part 125 by centrifugal force, and captures the target material trapped in the target material capturing part 125 From the means 128.

The target material collection step S36 may be performed such that the sample processing chip 110 rotates about the rotation axis X at a sixth rotation speed (for example, 5000 RPM) higher than the fifth rotation speed after the elution buffer solution loading step S35, And is rotated for a predetermined time (for example, 1 minute). Referring to FIG. 7G, the target material T separated from the capturing means 128 by the elution buffer solution in the target material collection step (S36) is stored in the target material chamber 170. FIG.

Referring again to FIG. 5, after the preprocessing step S30 is completed, the PCR execution step S40 is performed. In the PCR execution step (S40), the denaturation step, the combining step, and the elongation step are performed in order. The denaturing step is performed by locating each target material chamber 170 of the sample processing chip 110 in the first, fourth, seventh, and tenth heating zones 185a, 185d, 185g, and 185j that provide the temperature of the denaturation step . The bonding step is performed such that after the denaturation step is completed, the sample processing chip 110 is rotated at a certain angle so that each of the target material chambers 170 is heated in the second, fifth, eighth, and eleventh heating zones 185b, 185e, 185h, 185k. The elongating step is performed such that after the bonding step is completed, the sample processing chip 110 rotates at a certain angle so that each target material chamber 170 is heated to the third, sixth, ninth, and twelfth heating zones 185c, 185f, 185i, 185m.

Although the present invention has been described with reference to the above embodiments, the present invention is not limited thereto. It is to be understood that the above-described embodiments may be modified or changed without departing from the spirit and scope of the present invention, and those skilled in the art will recognize that such modifications and changes are also within the scope of the present invention.

100: PCR apparatus 110: sample processing chip
120: unit sample processing unit 125: target substance capturing unit
128: capturing means 130: sample chamber
135: sample introduction passage 140: washing buffer solution chamber
145: washing buffer solution introducing passage 146: outer switching curve portion
147: inner switching curve section 150: elution buffer solution chamber
155: eluting buffer solution introducing passage 156: first outside switching curve section
157: first inside switching curve portion 158: second inside switching curve portion
159: second inside switching curve section 160: discharge passage
165: waste liquid chamber 170: target material chamber
175: circulation passage 180: temperature control section
190:

Claims (14)

And a unit sample processing unit (120) spaced apart from the rotation center in the radial direction,
The unit sample processing unit includes:
A target material catching part 125 having a catching passage 126 having an inlet and an outlet and a catching means 128 filled in the catching passage;
A sample chamber 130 located radially inward of the target substance capturing portion and connected to an inlet of the trapping passage and providing a space in which a sample is stored;
A washing buffer solution chamber (140) located radially inward of the target material capturing part and providing a space for storing the washing buffer solution therein;
A washing buffer solution introducing passage (145) located between the washing buffer solution chamber and the inlet of the trapping passage;
An elution buffer solution chamber (150) located radially inward of the target substance capturing part and providing a space in which an elution buffer solution is stored; And
And an elution buffer solution introducing passage (155) located between the eluting buffer solution chamber and the entrance of the trapping passage,
The washing buffer solution introducing passageway includes an inner switching curve portion 147 for converting the flow of the washing buffer solution from the inside to the outside in the radial direction, And a second inside switching curve section (159) arranged in order along the flow direction, the first inside switching curve section (157) and the second inside switching curve section (159). The method according to claim 1,
Wherein the washing buffer solution introducing passage further comprises an outer switching curve portion (146) located on the upstream side relative to the inner switching curve portion and switching the flow of the washing buffer solution from the radially outer side to the inward side. Process Sample Processing Chip. The method according to claim 1,
The elution buffer solution introduction passage includes a first outside switching curve portion 156 located upstream of the first inside switching curve portion and switching the flow of the eluting buffer solution from the outside to the inside in the radial direction, Further comprising a second outer switching curve portion (158) located between the curved portions and switching the flow of the eluting buffer solution from the outside to the inside in the radial direction. The method according to claim 1,
Wherein the trapping channel connects the inlet and the outlet in a staggered configuration, the inlet and the outlet being located radially inward and outward, respectively. The method according to claim 1,
Wherein the unit sample processing unit further comprises a discharge passage (160) located radially outward of the target substance capturing unit and connected to an outlet of the trapping channel. The method of claim 5,
Wherein the unit sample processing unit further comprises a circulation passage (175) for connecting the discharge passage to the sample chamber, the wash buffer solution chamber, and the elution buffer solution chamber. The method according to claim 1,
Wherein the plurality of unit sample processing units are arranged along the circumferential direction. The method according to claim 1,
Wherein the capturing means is a silica-based material. The method according to claim 1,
Wherein the target substance capturing portion has a weir structure. A sample processing chip for rotary PCR process according to any one of claims 1 to 9; And
And a rotation driving unit for rotating the sample processing chip. The method of claim 10,
Further comprising a temperature regulator in which the sample processing chip is accommodated and a heating region for providing a required temperature for each step of the PCR process is disposed along the circumferential direction. A rotary PCR method using a sample processing chip for a rotary PCR process according to any one of claims 1 to 9,
Preparing a sample processing chip in which a sample is injected into the sample chamber, a washing buffer solution is injected into the washing buffer solution chamber, and an elution buffer solution is injected into the elution buffer solution chamber; And
And a preprocessing step of rotating the sample processing chip to perform a pretreatment on the sample,
The pre-
A target material capturing step of rotating the sample processing chip at a first rotation speed to introduce the sample into the target material capturing part;
Wherein the washing buffer solution is passed through the inside switching curve portion of the washing buffer solution introducing passage so that the eluting buffer solution is passed through the elution buffer solution A first introducing step of passing the first inner switching curve portion of the introduction passage;
A washing buffer solution loading step of increasing the rotational speed of the sample processing chip to a third rotational speed that is faster than the second rotational speed to introduce the washing buffer solution into the target substance capturing part;
A second introduction step of decelerating the rotation speed of the sample processing chip at a fourth rotation speed slower than the third rotation speed so that the elution buffer solution passes through the second inside transition curve portion of the elution buffer solution introduction passage; And
And an elution buffer liquid loading step of increasing the rotational speed of the sample processing chip to a fifth rotational speed that is faster than the fourth rotational speed so as to introduce the elution buffer solution into the target substance capturing part. The method of claim 12,
Wherein the second rotation speed and the fourth rotation speed are the same. The method of claim 12,
Further comprising a target material collecting step of collecting the target material by increasing the rotational speed of the sample processing chip to a sixth rotational speed that is higher than the fifth rotational speed after the elution buffer solution loading step.

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