KR20230047781A - Intergrated real-time pcr chip - Google Patents

Abstract

The present invention relates to an integrated real-time PCR chip, which comprises: a first substrate provided to accommodate a sample; a second substrate for heating the sample; and a third substrate disposed between the first substrate and the second substrate, wherein the first substrate includes a chamber for receiving the sample and a channel part provided to allow the sample to be introduced into the chamber. The integrated real-time PCR chip of the present invention can prevent a sample from leaking out during a PCR experiment by using a housing including a chamber and can minimize the use of tapes.

Description

Integrated real-time PCR chip {INTERGRATED REAL-TIME PCR CHIP}

The present application relates to an all-in-one real-time PCR chip.

Polymerase chain reaction (PCR) is an important technique for biological research because it tests for diseases with small amounts of DNA. It is used in a wide range of fields such as bacterial, fungal, criminal investigation, and diagnosis of infectious diseases caused by viruses. The DNA detection process requires four steps: DNA extraction, DNA amplification, electrophoresis, and gel image analysis. In general, PCR amplifies DNA and analyzes the results through several processes.

This process is time consuming and samples are potentially contaminated through several passive DNA detection steps. Real time PCR has been developed to overcome these disadvantages. Real-time PCR can detect the amount of nucleic acid amplification products in real time, analyze results in a shorter time than conventional methods, and minimize the possibility of contamination because there are no manual steps such as opening to monitor the reaction. A tube is required to transfer the to the detection device. Therefore, many studies on real-time PCR are being conducted and various devices are being sold.

Lab-on-a-Chip (LOC) is being developed to make biochemical processes cheaper and simpler. LOC is known as a technique capable of processing small amounts of samples. Therefore, it should be possible to lower the production cost and process a small amount of samples. Due to these demands, the development of microfluidic channels has become more active. Accordingly, channels using various materials have been developed, and experiments using readily available and flexible materials and easy-to-manufacture tapes are increasing. The tape is already available in a variety of colors and thicknesses, making it easier to fabricate than PDMS or PMMA.

Methods for detecting various fluorescence of the PCR chip generally include a method using a photodiode and a lens and a method using a camera including a CCD or CMOS sensor. The real-time PCR system has a chamber containing a sample for synthesizing DNA, and experiments are conducted at a high temperature of about 95 degrees during PCR amplification. Accordingly, air bubbles are generated in the sample, and it may be difficult to accurately measure fluorescence brightness.

Since real-time PCR experiments evaluate analysis and performance by fluorescence brightness values, accurate measurement may be difficult if air bubbles are generated in the channel during fluorescence brightness analysis. If an air bubble is created in the channel at the moment the fluorescence brightness is measured, the air bubble and its surroundings become dark. In addition, if the reagents to be amplified at the correct temperature according to the protocol order are out of the heating pattern due to air bubbles, the amplification efficiency greatly decreases.

In addition, the real-time PCR chip manufactured using a conventional tape has the advantage of being easily obtainable and easy to manufacture, as described above. However, various disadvantages may occur during production in the method of manufacturing by stacking several tapes. Tapes of different shapes must be processed, and disruptions occur when raw materials are out of stock or supply is interrupted. The most fatal disadvantage is that during PCR amplification at a high temperature of 95 degrees, the tapes attached to each other are deformed, and the adhesiveness of the tapes attached to each other deteriorates, causing the sample to leak into the chamber made by processing the tapes, resulting in normal experiments. A problem occurred that made this impossible.

In addition, each hole through which the sample is injected and withdrawn must be blocked in order to maintain a constant temperature or escape of the sample during PCR amplification. To this end, several methods are being used. In general, a tube type is used to contain the sample, and an integral cover is attached, but there are not many cases in which a chip type is used, so much research is needed for this.

The background technology of the present application is disclosed in Korean Patent Publication No. 10-2015-0094842.

The present invention is to solve the above-mentioned problems of the prior art, to prevent the leakage of the sample by using a housing including a chamber during PCR experiments, to minimize the use of tape, and to be integrally provided through a simple coupling structure An object of the present invention is to provide an all-in-one real-time PCR chip.

In addition, after the sample inlet and outlet are made in the PCR chip housing, an integral cover is included, and the shape of the chamber and channel is changed to collect air bubbles generated at high temperatures in air pockets, greatly increasing amplification efficiency and providing stable experimental results. An object of the present invention is to provide an all-in-one real-time PCR chip.

However, the technical problem to be achieved by the embodiments of the present application is not limited to the technical problems described above, and other technical problems may exist.

As a technical means for achieving the above technical problem, an integrated real-time PCR chip includes a first substrate provided to accommodate a sample; a second substrate for heating the sample; and a third substrate disposed between the first substrate and the second substrate, wherein the first substrate may include a chamber accommodating the sample and a channel portion provided to allow the sample to flow into the chamber. there is.

In addition, the chamber includes an air pocket formed in a shape in which the width increases toward the lower direction and an amplification unit extending from the other end of the air pocket toward the other side but having a cross-sectional area larger than that of the air pocket, The channel unit may include a first channel extending upward and downward to introduce a sample from an upper side, and a second channel bent upward from a lower side of the first channel and extending to a lower side of the amplifying unit.

In addition, the channel unit may include an inlet for injecting the sample and an outlet for discharging the sample, and the integrated real-time PCR chip may further include an integrated cover provided to block the inlet and the outlet. .

In addition, the sample may be introduced through the inlet and the first channel, and may be discharged through the top of the air pocket after PCR is completed.

In addition, the integrated real-time PCR chip may perform PCR in a vertical state.

In addition, the second substrate may include a heater for heating the sample; And it may include a temperature measuring unit for measuring the temperature of the sample.

In addition, the third substrate may be provided as an adhesive member interposed between the first substrate and the second substrate so that the first substrate and the second substrate are attached to each other.

In addition, the integrated real-time PCR chip may further include a fourth substrate accommodating the first to third substrates, and the fourth substrate may be provided as a housing.

In addition, the first substrate and the fourth substrate may be coupled to each other, and the integrated real-time PCR chip may be integrally provided.

The above-described problem solving means are merely exemplary and should not be construed as intended to limit the present disclosure. In addition to the exemplary embodiments described above, additional embodiments may exist in the drawings and detailed description of the invention.

According to the above-described problem solving means of the present application, the integrated real-time PCR chip according to an embodiment of the present application includes a first substrate and a fourth substrate provided to be coupled to each other, so that the sample bursts or leaks due to high temperature By preventing this, safer and more accurate experiments may be possible.

In addition, according to the above-described problem solving means of the present application, the first substrate includes a chamber and a channel portion for accommodating the sample, thereby eliminating the manufacturing step of punching or cutting the tape in the shape of a microfluidic channel during the PCR amplification experiment. It can be made simply and inexpensively.

In addition, according to the above-mentioned problem solving means of the present application, by including an air pocket formed in a shape in which the width increases as the chamber of the first substrate goes downward, air bubbles generated during PCR are collected in a separate space and sample can be efficiently accommodated, and it is possible to prevent reduction of internal brightness of the channel due to air bubbles during sample analysis.

In addition, according to the above-mentioned problem solving means of the present application, by including an integral cover provided to block the inlet and outlet on the first substrate, the experimenter's convenience is increased during the PCR amplification experiment and the temperature is stably maintained to detect nucleic acid amplification. can do.

However, the effects obtainable herein are not limited to the effects described above, and other effects may exist.

1 is a schematic exploded perspective view of an integrated real-time PCR chip according to an embodiment of the present application.
2 is a schematic configuration diagram (perspective view) of an integrated real-time PCR chip according to an embodiment of the present application.
3 is a schematic diagram of a first substrate of an integrated real-time PCR chip according to an embodiment of the present disclosure.
4 is a schematic diagram for explaining a chamber and a channel portion of a first substrate of an integrated real-time PCR chip according to an embodiment of the present disclosure.
5 is a schematic diagram for explaining an integrated real-time PCR chip according to an embodiment of the present application.
6 is a schematic configuration diagram for explaining an integrated real-time PCR chip according to an embodiment of the present application.
7 is an exploded configuration diagram for illustratively explaining an implementation example of an integrated real-time PCR chip according to an embodiment of the present application.
8 is an assembly completion diagram for illustratively explaining an embodiment of an integrated real-time PCR chip according to an embodiment of the present disclosure.
9 is a diagram schematically showing the results of PCR amplification experiments using an integrated real-time PCR chip according to an embodiment of the present application.
10 is a diagram schematically showing photographs of changes in fluorescence brightness before and after PCR amplification using an integrated real-time PCR chip according to an embodiment of the present application.

Hereinafter, embodiments of the present application will be described in detail so that those skilled in the art can easily practice with reference to the accompanying drawings. However, the present disclosure may be implemented in many different forms and is not limited to the embodiments described herein. And in order to clearly describe the present application in the drawings, parts irrelevant to the description are omitted, and similar reference numerals are attached to similar parts throughout the specification.

Throughout this specification, when a part is said to be "connected" to another part, this includes not only the case of being "directly connected" but also the case of being "electrically connected" with another element in between. do.

Throughout the present specification, when a member is referred to as being “on,” “above,” “on top of,” “below,” “below,” or “below” another member, this means that a member is located in relation to another member. This includes not only the case of contact but also the case of another member between the two members.

Throughout the present specification, when a part "includes" a certain component, it means that it may further include other components without excluding other components unless otherwise stated.

The present application relates to a functional change of a channel provided with an air pocket for removing air bubbles generated due to high temperature during PCR amplification in a Polymerase Chain Reaction Chip (hereinafter referred to as 'PCR chip'). . During the experiment, the sample does not flow fluidly, and amplification proceeds while the sample is contained in a channel with an air pocket at one end. Accurate fluorescence measurement is possible by separately collecting the air bubbles in the channel to the upper part of the air pocket containing the sample.

Hereinafter, an integrated real-time PCR chip (hereinafter referred to as 'this PCR chip') according to an embodiment of the present application will be described.

1 is a schematic exploded perspective view of an integrated real-time PCR chip according to an embodiment of the present application, FIG. 2 is a schematic configuration diagram (perspective view) of an integrated real-time PCR chip according to an embodiment of the present application, and FIG. It is a schematic diagram of a first substrate of an integrated real-time PCR chip according to an embodiment. 4 is a schematic diagram for explaining a chamber and a channel portion of a first substrate of an integrated real-time PCR chip according to an embodiment of the present disclosure.

For example, according to one embodiment of the present application, the PCR chip 1 has a four-layer structure consisting of a printed circuit board (PCB), a plastic cover, a plastic housing, and a transparent double-sided tape connecting the housing and the PCB. PCB (Printed Circuit Board) is the basic material of a PCR chip. A printed circuit board (PCB) can include a heater pattern that can be heated and a thermistor attached to the bottom. After fabricating this PCR chip (1) using a PCB and a plastic cover, PCR testing and gel electrophoresis testing can be performed.

LEDs can be used as illumination for measuring the amount of fluorescence. When using a glossy green PCB, the overall signal-to-noise ratio is reduced because the PCB reflects light and acts as noise in the process of measuring and analyzing fluorescence brightness. Therefore, by replacing the previous glossy green PCB with a matte black PCB and printing a white silk legend on the heating pattern part under the channel part, the brightness of the fluorescence changed in real time during the PCR process can be easily distinguished.

In addition, the PCR of the present application may be performed by standing the PCR chip 1 vertically. PCR is performed by standing the PCR chip 1 vertically, and by providing a separate space (air pocket 11a) for collecting air bubbles, the heated hot air gathers upward and the fluorescence brightness can be measured more efficiently.

In addition, the PCR chip 1 is provided as an integral type, and a more safe and accurate experiment can be performed by preventing the sample from bursting or leaking due to high temperature.

Referring to FIGS. 1 and 2 , the PCR chip 1 includes a first substrate 10 , a second substrate 20 and a third substrate 30 .

Referring to FIGS. 1 and 3 , the first substrate 10 may be provided to accommodate a sample. For example, the sample may be a liquid sample synthesizing DNA.

For example, the first substrate 10 (housing) is made of a plastic material and may be coated with a matte black coating to block light reflection. Also, for example, the first substrate 10 may be made of a transparent material for light transmission and detection with respect to a portion provided with the chamber 11 described below. For example, the transparent material may be any material that transmits light, and glass or synthetic resin may be used. For example, the synthetic resin may include transparent silicone, transparent orethane, transparent PVC, transparent polypropylene, transparent polycarbonate, and mixtures thereof.

In addition, the first substrate 10 may be provided as a cover film. The cover film may include polycarbonate. For example, the first substrate 10 may be a polycarbonate film having a thickness of 500 μm. Since the first substrate 10 is formed of a polycarbonate film having a thickness of 500 μm, it is possible to prevent expansion of the cover film under high temperature conditions during PCR. Since the first substrate 10 is made of polycarbonate, damage to the surface of the film can be minimized and durability of the film can be improved. In addition, since the first substrate 10 is made of a polycarbonate film, a change in brightness can be clearly observed during fluorescence detection.

In addition, the first substrate 10 may accommodate a sample for synthesizing DNA to perform PCR. For example, the first substrate 10 may be a microfluidic channel. For example, the first substrate 10 may be formed by being laminated to include at least one of a plastic material, a cover film, a transparent material, a silicon material, and a dry film resist (DFR), but is not limited thereto no. Since the thermal conductivity of DFR is very low compared to that of silicon, it can exhibit excellent performance in keeping a sample accommodated in the first substrate 10 warm. As another example, for a silicon material, a space for accommodating a sample may be formed inside the second substrate 20 by using an etching process using a mask pattern. For DFR, a space for accommodating a sample may be formed using an exposure process and a development process. The first substrate 10 may receive a sample introduced from the sample inlet (inlet 13a) and perform a polymerase chain reaction (hereinafter referred to as PCR).

On the other hand, when PCR is performed, air bubbles generated in a high-temperature section of about 95 degrees are located in the middle of the channel, and thus there is a problem that the shadow of the air bubble portion may decrease overall brightness during fluorescence analysis of the image. In addition, there is a problem that a large amount of the sample may be lost as air bubbles push the sample accommodated inside the channel to the outside.

Accordingly, in the present application, the first substrate 10 may include a space for injecting a sample, a space for accommodating the sample, and a space for collecting high-temperature air bubbles. In other words, the first substrate 10 may have air pockets 11a on the top of which high-temperature air bubbles can be collected when PCR is performed by standing the PCR chip 1 vertically. In order to solve the occurrence of air bubbles due to high temperature in measuring fluorescence brightness, the first substrate 10 may include a modified U-shaped microfluidic channel by manufacturing a cone-shaped space in the existing channel. Also, the first substrate 10 may be made of an epoxy component.

Referring to FIGS. 1 , 3 and 4 , the first substrate 10 may include a chamber 11 and a channel unit 12 . The chamber 11 of the first substrate 10 may accommodate a sample, and the chamber 11 may include an air pocket 11a and an amplification unit 11b.

The air pocket (11a) may be formed in a shape in which the width increases toward the lower direction. For example, the air pocket 11a may be formed in a cone shape. Also, for example, the width of the air pocket (11a) may be greater than the width of the channel portion (12). The large volume of the air pocket 11a can sufficiently collect a large amount of air generated during PCR. The upper part of the air pocket (11a) may be formed with a relatively narrow width compared to the lower part. In addition, the air pocket 11a may be a region provided so that high-temperature air bubbles can be collected on the upper part of the first substrate 10 (channel) when PCR is performed by standing the PCR chip 1 vertically. By providing the air pocket 11a inside the first substrate 10, air bubbles generated during PCR are collected in a separate space, and the loss of the sample in the first substrate 10 (channel) due to the air bubbles is fundamentally prevented. By preventing this, the sample can be accommodated efficiently. In addition, when the fluorescence detection test is performed with the PCR system by having the air pocket 11a, it is possible to confirm that the success of nucleic acid amplification and the change in fluorescence brightness are stable, shorten the experiment time, and perform PCR with low noise. there is.

In addition, referring to FIGS. 3 and 4 , the amplifying unit 11b extends from the other end of the air pocket 11a toward the other side and may be formed to have a cross-sectional area larger than that of the air pocket 11a. For example, the amplification unit 11b may refer to a region including a reagent supplied from the channel unit 12 . In addition, the amplification unit 11b may be a region that is photographed to compare fluorescence brightness in the process of performing PCR. Illustratively, referring to FIG. 3 , the amplifier 11b may refer to a space containing reagents. The amplification unit 11b calculates the light reflected from the irradiated light as a voltage and detects the fluorescence brightness of the sample in the amplification unit 11b to confirm DNA amplification.

The channel unit 12 may be provided to allow a sample to flow into the chamber 11 . For example, the channel unit 12 may be a passage through which a reagent (sample) is filled in the amplification unit 11b. The channel unit 12 may include a first channel 12a extending in an upward and downward direction so that a sample is introduced from the upper side. In addition, the channel unit 12 may include a second channel 12b that is bent upward from a lower portion of the first channel 12a and extends to a lower portion of the amplifier 11b. The channel unit 12 may include an inlet 13a for injecting a sample and an outlet 13b for discharging the sample. For example, a sample may be introduced (injected) from the inlet 13a and the upper end of the first channel 12a may receive the sample injected from the inlet 13a, and the second channel 12b may be provided with the first channel ( It can serve as a passage through which the sample moved along 12a) can be moved into the amplification unit 11b. In addition, after the PCR is completed, it can be discharged to the top of the air pocket (11a). For example, an outlet 13b may be formed on top of the air pocket 11a. In most PCR experiments, air bubbles are not generated in the channel part 12, and even if they are generated, they disappear or move to the chamber during amplification.

Illustratively, referring to FIG. 4 , a sample is supplied through the first channel 12a and the inside of the amplifier 11b along the curved surfaces of the first channel 12a and the second channel 12b by capillary action. The sample is filled in, and air generated during PCR is immediately discharged into the air pocket 11a, so that shadows of air bubbles in the amplification unit 11b can be fundamentally prevented.

In addition, by including air pockets 11a in the first substrate 10, air bubbles generated during PCR are collected in a separate space, and loss of samples in the amplification unit 11b due to air bubbles is fundamentally prevented. By doing so, the sample can be received efficiently.

Referring to FIG. 1 , the PCR chip 1 may include an integral cover 50 provided to block the inlet 13a and the outlet 13b. For example, the integrated cover 50 may be made of a silicon material to block holes of the inlet 13a and the outlet 13b of the channel unit 12 for injecting and collecting liquid samples.

5 is a schematic diagram for explaining an integrated real-time PCR chip according to an embodiment of the present application, and FIG. 6 is a schematic configuration diagram for explaining an integrated real-time PCR chip according to an embodiment of the present application.

Meanwhile, the second substrate 20 of the PCR chip 1 may heat the sample. For example, the second substrate 20 may be provided as a PCB substrate. For example, the second substrate 20 may be provided as a matte black PCB substrate. Also, for example, the second substrate 20 may be formed to a thickness of 0.2T - 200μm. By providing the second substrate 20 with a matte black PCB, it is possible to minimize the reflectance by the PCB and prevent the reflected light from acting as noise, thereby reducing the ratio of noise generation to the total signal.

Referring to FIGS. 5 and 6 , the second substrate 20 may include a heater 21 for heating the sample and a temperature measuring unit for measuring the temperature of the sample. The heater 21 may heat the sample. The heater 21 may heat the sample provided in the amplification unit 11b to perform a PCR reaction. For example, the heater 21 may be a thermoplate. The thermoplate is composed of platinum (Pt), silver (Ag), etc., or formed by coating a conductive material such as platinum (Pt), silver (Ag), titanium (Titanium, Ti), etc. by sputtering. It can be.

Also, the temperature measuring unit may be a thermocouple. The temperature measurement unit may be provided as a thin thermal couple line to measure the temperature of the reagent inside the amplification unit 11b of the first channel 10 . The temperature measuring unit may measure the temperature of the sample and the chip, compare the measured internal temperature of the sample with the temperature of the chip, and correct the internal temperature of the sample to reach a target temperature of the chip. For example, driving of the heater 21 may be controlled based on the temperature measured by the temperature measuring unit.

Also, the second substrate 20 may include a heater pattern. The heater pattern may be a region provided for heating. The heater pattern may be formed in an area corresponding to the area of the amplifier 11b included in the first substrate 10 . Illustratively, by providing a heater pattern with an annotation legend printed on a heating pattern portion at the lower end of the first substrate 10, it is possible to easily distinguish the brightness of fluorescence changed in real time while performing the PCR process. However, it is not limited thereto, and the heater pattern may be provided by printing a white silk legend.

Also, for example, referring to FIGS. 5 and 6 , the second substrate 20 may be provided to enable connection with a control unit for controlling the heater 21 and the temperature measuring unit. Accordingly, the control unit may control the heater 21 to heat the sample inside the amplification unit 11b or determine the temperature measured through the temperature measuring unit.

7 is an exploded configuration diagram for exemplarily explaining an implementation example of an integrated real-time PCR chip according to an embodiment of the present application, and FIG. 8 is an example of an implementation example of an integrated real-time PCR chip according to an embodiment of the present application. It is an assembly completeness for explanation.

Referring to FIGS. 1 and 7 , a third substrate 30 may be disposed between the first substrate 10 and the second substrate 20 . The third substrate 30 may be provided as an adhesive member to be interposed between the first substrate 10 and the second substrate 20 so that the first substrate 10 and the second substrate 20 are attached to each other. For example, the third substrate 30 may be provided with an adhesive member including tape for use in bio experiments, or more specifically, may be provided with a transparent double-sided tape.

Also, referring to FIGS. 1 , 7 and 8 , the PCR chip 1 may include a fourth substrate 40 . The fourth substrate 40 may accommodate the first substrate 10 to the third substrate 30 and may be provided as a housing. For example, the fourth substrate 40 is made of a plastic material and may be coated with a matte black coating to block light reflection.

Referring to FIGS. 1, 2 and 8, the first substrate 10 and the fourth substrate 40 may be coupled to each other, and accordingly, the PCR chip 1 may be integrally provided. can For example, the first substrate 10 and the fourth substrate 40 may be bound (coupled) using a hook method.

In this PCR chip (1), the amplification part (11b) and the channel part (12) are attached to the first substrate (10, housing) made of plastic instead of the existing method of manufacturing and attaching the amplification part and the channel part using tape. By including it, it can be manufactured more simply and inexpensively by eliminating the manufacturing step of punching or cutting the tape into the shape of a microfluidic channel during PCR amplification experiments.

In addition, this PCR chip 1 is different from the conventional adhesive method using a plurality of tapes, and the first substrate 10 and the fourth substrate ( 40) by using a hook method, it is possible to prevent the liquid sample from bursting or leaking due to high temperature.

In addition, the present PCR chip 1 includes an integral cover 50 covering the inlet 13a and the outlet 13b on the first substrate 10 to increase the convenience of the experimenter and stably maintain the temperature during the PCR amplification experiment. Thus, the amplification of nucleic acids can be detected.

9 is a diagram schematically illustrating PCR amplification test results using an integrated real-time PCR chip according to an embodiment of the present application, and FIG. 10 is a fluorescence brightness change before and after PCR amplification using an integrated real-time PCR chip according to an embodiment of the present application. It is a schematic representation of the picture.

Illustratively, the results of PCR amplification experiments can be converted into graphs through an Excel program by quantifying the brightness of experimental data (fluorescent photographs) using a program called IMAGE J. For example, referring to FIG. 9 , the results of PCR amplification experiments using the present PCR chip 1 can show stable results in all experiments.

In addition, referring to FIG. 10, it can be confirmed that there are almost no air bubbles in the microfluidic channel during the PCR amplification experiment using the present PCR chip (1).

According to another embodiment of the present application, the PCR chip 1 may be controlled by a PCR chip control system. The PCR chip control system is basically responsible for two categories of functions: biochemical function and user interface function. The biochemical function can drive the PCR protocol and control the temperature of the chip according to the temperature set in the PCR protocol. Also, this PCR chip 1 can be controlled by a PCR protocol. The PCR protocol can be pre-set to run at a specified temperature for a specified period of time. Heating and cooling of the PCR chip 1 may be designed to perform temperature sensing using a thermistor, and heating and cooling by a heater and a fan. The PCR protocol may control the PCR chip 1 through a series of operations instructing the system to maintain a specific temperature for a certain period of time. After driving the heater or fan in the real-time PCR system, heating or cooling the chip is performed according to the set temperature, and a process to be processed can be determined later. In addition, the PCR chip control system may include a user interface function for a user including functions such as file management and editing in addition to a biochemical function of processing a process for temperature control.

The temperature of the PCR chip 1 may increase or decrease at a rate of about 10° C./s. Temperature control settings for performing the PCR protocol can be set within an error range of ±0.5 °C. In addition, the PCR protocol may perform processing shorter than a preset time (eg, 50 ms) in order to control the temperature error. When the work is processed in real time and viewed as a standard criterion for functional division that must be met, the PCR protocol (not shown) repeats the second protocol step of heating from 72 ° C to 95 ° C, so that the standard is not violated. The PCR protocol process can be performed on the host PC.

The above description of the present application is for illustrative purposes, and those skilled in the art will understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present application. Therefore, the embodiments described above should be understood as illustrative in all respects and not limiting. For example, each component described as a single type may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined form.

The scope of the present application is indicated by the following claims rather than the detailed description above, and all changes or modifications derived from the meaning and scope of the claims and equivalent concepts thereof should be construed as being included in the scope of the present application.

1: All-in-one real-time PCR chip
10: first substrate
11: chamber
11a: air pocket
11b: amplification unit
12: channel unit
12a: first channel
12b: second channel
13a: inlet
13b: Outlet
20: second substrate
21: heater
30: third substrate
40: fourth substrate
50: integral cover

Claims (9)

In the PCR chip,
a first substrate provided to accommodate a sample;
a second substrate for heating the sample;
A third substrate disposed between the first substrate and the second substrate,
The first substrate,
The integrated real-time PCR chip comprising a chamber for accommodating the sample and a channel unit provided so that the sample can flow into the chamber. According to claim 1,
The chamber includes an air pocket formed in a shape in which the width increases toward the lower direction and an amplification part extending from the other end of the air pocket toward the other side and having a cross-sectional area larger than that of the air pocket,
The channel unit includes a first channel extending upward and downward to allow the sample to flow in from the upper side, and a second channel bent upward from the lower side of the first channel and extending to the lower portion of the amplification unit. According to claim 2,
The channel part,
Including an inlet for injecting the sample and an outlet for discharging the sample,
The integrated real-time PCR chip,
The integral real-time PCR chip further comprising an integral cover provided to block the inlet and the outlet. According to claim 3,
The sample is introduced through the inlet and the first channel and discharged through the upper part of the air pocket after PCR is completed, an integrated real-time PCR chip. According to claim 1,
The integral real-time PCR chip is an integral real-time PCR chip in which PCR is performed in a vertical state. According to claim 1,
The second substrate,
a heater for heating the sample; and
All-in-one real-time PCR chip comprising a temperature measuring unit for measuring the temperature of the sample According to claim 1,
The third substrate,
An integrated real-time PCR chip interposed between the first substrate and the second substrate and provided as an adhesive member so that the first substrate and the second substrate are attached to each other. According to claim 1,
The integrated real-time PCR chip further includes a fourth substrate accommodating the first to third substrates,
The fourth substrate is provided as a housing, integrated real-time PCR chip. According to claim 8,
The first substrate and the fourth substrate are provided to be coupled to each other,
The integral real-time PCR chip, wherein the integral real-time PCR chip is provided integrally.

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