KR20200061686A - Integrated Chip for DNA Analyzer - Google Patents

Abstract

The present invention relates to a chip device for a portable DNA analyzer. The chip device includes: an upper plate (10); a DNA film chip (20) disposed in such a manner that it may face the bottom of the upper plate (10); and a lower plate (30) fixed to the lower half portion of the upper plate in such a manner that the rear end portion of the DNA film chip (20) may be interposed between the upper plate (10) and the lower plate (30). It is possible to carry out a series of processes, including extracting and amplifying DNA genetic materials from a subject, separating multiple specific genes at once by using a capillary electrophoresis system and reading the result from the separated image, rapidly on the spot through an integral type device system. In addition, gene amplification, electrophoresis and analysis result reading can be carried out continuously, thereby providing an effect of reducing a time required for amplification, a time required for electrophoresis and a time required for analysis result reading.

Description

Integrated chip device for analysis {Integrated Chip for DNA Analyzer}

The present invention relates to a chip device for DNA analysis, and more particularly, to an integrated chip device for DNA analysis configured to be able to perform the entire process in one chip easily and conveniently in a place where DNA analysis is required in real time.

In general, DNA collected from a subject is analyzed using a polymerase chain reaction (PCR) method, and in order to proceed with such a PCR method, a thermal cycler device capable of amplifying the collected DNA and a thermal cycler equipment are collected. Control equipment is needed to control the analysis of a DNA.

However, the real-time PCR equipment to which the conventional PCR method is applied is developed for a desktop and has a very large size, and is configured to operate in connection with an external computer for data processing, and the internal structure is designed with low spatial efficiency. Therefore, it can be said that there are many inefficient and unnecessary parts for use in the actual field.

In particular, if you want to analyze the DNA of a suspected foot-and-mouth disease animal in order to determine a hereditary infectious disease such as foot-and-mouth disease, or you need to analyze this DNA by moving it to a place where real-time PCR equipment is installed, you will leave the area and secondary infection will occur. In addition to being concerned, serious problems may arise that are difficult to manage according to the travel time.

In addition, the molecular biology test, which is a test method that can be diagnosed early by confirming the presence or absence of direct factors such as genetic material of an infectious agent, has a risk of false positive results due to the tester or contamination.

Accordingly, in recent years, WHO has emphasized the need for on-site inspection for the diagnosis of infectious diseases, and a system for on-site inspection using molecular biological methods has been steadily developed since about 10 years ago.

Moreover, with the advent of lab-on-a-chip (LOC) technology that allows all molecular diagnostic procedures that consist of the extraction of genetic material from a sample, gene amplification reactions, and diagnosis of amplification, etc., in one system, field diagnosis Research on the high-efficiency and high-sensitivity system is being actively conducted.

As a result of this research, the strip sensor module for the molecular diagnostic field inspection device disclosed in Korean Patent Registration No. 10-1767978, the gene quantitative detection real-time PCR and genetic analysis DNA microarray disclosed in Korean Patent Registration No. 10-1302353 were used. There are automatic molecular diagnostic equipment that can be performed in combination.

However, in the case of such a conventional molecular diagnosis field inspection device, the DNA chip and measurement device configuration is generally complicated, so the cost of manufacturing the product is high, and the cost of using a single test is also high. To confirm the result of the PCR product, Since it has to be moved to a separate measuring device, there is a disadvantage in that measurement reliability due to contamination in the process of moving is low, and since gene extraction, amplification, separation, and reading are not made in one system, initial judgment is possible to some extent, but the final result is There is a disadvantage that it is not suitable for on-site inspection because it takes a long time to obtain.

Korean Registered Patent No. 10-1767978 Korean Registered Patent No. 10-1302353

The present invention has been devised to solve the above-mentioned problems, and the object of the present invention is to amplify the DNA genetic material extracted from a sample using a single integrated chip device, and to use a capillary electrophoresis device to generate multiple specific genes. It is to provide an integrated chip device for DNA analysis configured to perform a series of processes of separating at a time and reading a result from a separated image in real time in the field.

In order to solve the above problems, the integrated chip device for DNA analysis according to the present invention,

It is characterized in that it comprises a gene detection and analysis module that is affixed to and fixed on a sample deck, and is subjected to amplification and electrophoresis by introducing a DNA sample.

In addition, in the integrated chip device for DNA analysis according to the present invention,

The gene detection analysis module,

Top plate 10;

A DNA film chip 20 disposed to correspond to a lower portion of the top plate 10; And

It is characterized in that it is fixed to the lower half of the upper plate, and is composed of a lower plate (30) with a rear end of the DNA film chip (20) interposed between the upper plate (10).

In addition, in the integrated chip device for DNA analysis according to the present invention,

The top plate 10,

From the upper end, the air injection port 11, the sample injection port 12, the through hole 3, the gel solution injection port 14 and the DNA collection port 15 are sequentially formed in a plate shape,

The DNA film chip 20,

The electrophoretic section 26 including the air injection section 21 and the sample injection section 22, the gene amplification section 23 and the gel solution injection section 24 and the DNA collection section 25 is formed of fine capillaries,

The lower plate 30,

Characterized in that the contact point to which electricity can be applied is provided at the upper and lower ends to separate the nucleic acid of the amplified sample from the electrophoretic section 26, and to form an analysis reading section together with the upper plate 10. Integrated chip device for DNA analysis.

do.

In addition, in the integrated chip device for DNA analysis according to the present invention,

The rear end portion of the DNA film chip 25 is coupled between the upper plate 20 and the lower plate 27 and is characterized by being fixed with a fixed coupling band 40.

In addition, in the integrated chip device for DNA analysis according to the present invention,

The DNA analysis chip device,

It is configured to continuously perform gene amplification, electrophoresis, and analysis reading, and is characterized by reducing the time required for amplification, electrophoresis, and analysis reading.

The integrated chip device for DNA analysis according to the present invention extracts and amplifies DNA genetic material from a sample through an integrated device system, separates a plurality of specific genes at once using a capillary electrophoresis device, and separate images There is an effect that can quickly perform a series of processes to read the results from the field.

In addition, the integrated chip device for DNA analysis according to the present invention is configured to continuously perform gene amplification, electrophoresis and analytical reading, thereby reducing the time required for amplification, electrophoretic time, and analysis reading. have.

1 is a perspective view briefly showing the internal structure of an integrated chip device for DNA analysis according to an embodiment of the present invention,
Figure 2 is an exploded perspective view of Figure 1;

Hereinafter, preferred embodiments of the integrated chip device for DNA analysis according to the present invention will be described in more detail with reference to the accompanying drawings.

1 is a perspective view schematically showing an internal structure of an integrated chip device for DNA analysis according to an embodiment of the present invention, and FIG. 2 is an exploded perspective view of FIG. 1.

1 and 2, the integrated chip device 1 for DNA analysis according to the present invention amplifies a DNA sample extracted from a collected sample to a desired DNA region, and amplifies the sample to a gel solution. It is an integrated chip that moves according to the voltage applied therein to separate, and continuously processes the process of analyzing and reading the separated sample image, and consists of a gene detection and analysis means mounted on a sample deck (not shown). do.

One side of the sample deck (not shown) is connected to a sliding motor fixedly connected to the inside of the front part of the case of a miniaturized DNA analysis device to enable on-site inspection, and is slid in the front-rear direction. A rectangular groove having a predetermined width and length is formed.

Gene detection and analysis means is that the extracted DNA sample is amplified and electrophoresis is made, consisting of a top plate 10, a DNA film chip 20 and a bottom plate 30.

The upper plate 10 is a transparent rectangular plate shape having a predetermined width and length, from the upper end air inlet 11, sample inlet 12, through hole 13, gel solution inlet 14 and DNA collection port 15 Are sequentially formed and extended to form the second half of a certain length. The thickness of the top plate is made of approximately 3mm, but it is of course not limited thereto.

The DNA film chip 20 comprises a linear electrophoresis unit (26) comprising an air injection unit (21), a sample injection unit (22), a gene amplification unit (23), and a gel solution injection unit (24) and a DNA collection unit (25). ) Is formed of a fine capillary tube, which is disposed to correspond to the rear surface of the upper plate, and the lower end of the film chip is coupled between the upper plate and the lower plate and fixed with a fixed coupling band. Although the thickness of the film chip is approximately 430, it is of course not limited to this.

The air injection portion 21 and the sample injection portion 22 are provided with air injection holes and sample injection holes communicating with the air injection holes 11 and the sample injection holes 12 of the top plate 20, respectively. It allows the remaining bubbles and excess samples contained in the chip to be discharged, and the DNA sample extracted from the sample can be introduced.

At this time, a standard material serving as a reference marker is loaded into one of the fine capillaries of the sample injection part, and a sample is loaded into the remaining eight capillaries.

The gene amplification unit 23 amplifies a plurality of extracted DNAs to a desired DNA region, and can be heated or cooled to a temperature controlled according to the DNA sample by a heating and cooling device (not shown) for efficient amplification. Do.

The linear electrophoresis unit 26 is a microchip-based microcapillary tube formed by injection molding, and is formed to be continuous to the gene amplification unit 23.

The gel solution is injected into the injection hole of the gel solution injection part 24 communicating with the gel solution injection port 14 so that the nucleic acid of the amplified DNA sample is separated in the injected gel solution.

At this time, the linear electrophoresis unit 26 is also provided with a sample having a DNA to be measured for reference and a reference marker formed of a capillary of the same size equipped with a gel solution on the side, which is the same as the amplified sample of the sample. Allow to separate under conditions.

By providing the reference marker on the side as described above and separating the sample at the same time, the analysis difference caused by the conditional variation does not occur in the external environmental factors and separation process, thereby enabling more precise analysis and reading.

Here, a polyacrylamide gel or agarose gel may be used as a gel solution as a support, and the gel has functions of preventing convection, protecting a sample, and molecular sieve.

In addition, it is a matter of course that nucleic acids separated from the DNA collection unit 25 can be collected.

The lower plate 30 is a transparent rectangular plate shape having a predetermined width and length, and is provided with a contact point to which electricity can be applied to the upper and lower ends to separate the nucleic acid of the sample amplified by the electrophoresis section 26 by the applied electricity. It is possible to make a comparative measurement, fix a film chip interposed between the top plate and a plurality of linear images separated from the linear electrophoresis unit 26, and form an analysis reading unit for analyzing and reading compared to a reference marker. .

The size from the front end to the bottom of the lower plate 30 has a length corresponding to a surface extending from the gel solution inlet 14 of the upper plate 10 to the rear end of the upper plate, and the thickness of the lower plate 30 is approximately 5 mm. It is made, but of course not limited to this.

The fixed coupling band 40 is to fix the rear portion of the film chip interposed between the upper plate and the lower plate and the upper plate 10, and has coupling protrusions formed inside.

The inner coupling protrusion not only increases the bonding strength between the upper plate and the lower plate and the film chip, but also connects the gel solution inlet 24 to prevent the solution from leaking to the outside during the process.

The common area of the lower panel corresponding to the upper panel is an analysis reading unit, and analysis and reading are performed by comparing a plurality of linear images separated from nucleic acids in the electrophoretic unit 26 with a reference marker. For such an analysis, an imaging device for photographing the analysis reading unit may be provided at the upper portion and a light guide plate serving as a lower illumination may be configured at the lower portion.

The light guide plate may include a blue LED or an ultraviolet LED.

Integral chip device 1 for DNA analysis according to the present invention, amplifies a DNA sample extracted from a sample collected by simply moving anywhere, anytime, to a desired DNA region, and the amplified sample is a voltage applied in the gel solution It can be separated by moving according to, and the process of analyzing and reading the separated sample image can be continuously processed in real time.

Hereinafter, the operation of the integrated chip device 1 for DNA analysis according to the present invention will be described.

Air is supplied to the air inlet for each air inlet to remove moisture and remove excess samples.

Then, the prepared DNA sample is injected into the capillary of the sample injection part via a sample injection port (not shown) or through a sample injection port using a dispenser or through activation of an input operation button.

When the injected DNA sample is rapidly moved to the gene amplification section by a capillary phenomenon, amplification is performed while the gene amplification section is heated and cooled through a heating and cooling device.

The amplified sample is injected into the capillaries of the electrophoretic section 26 connected to communicate with the respective capillaries, and the gel solution is introduced through the gel solution inlet.

Next, when a DC voltage is applied to the electrophoretic unit, charged particles move in the injected gel, and the amplified DNA sample is separated and moved for each charged particle. This is because components such as nucleic acids are separated because they move according to the type of charged particles and at the same time have different transfer rates depending on the molecular weight. At this time, the movement speed is fast, such as the gel solution and the gene rapidly moving due to the capillary phenomenon according to the microchip-based capillary structure, and accordingly, the movement is rapidly spread and thus analysis is possible in a short time.

Next, a plurality of linear images measured separately from the electrophoretic unit are photographed using an imaging device and an LED light, compared with a reference marker through an internal analysis program, and analyzed and read, and the analyzed result is displayed on the display device or It may be provided to be shown in a separate play means.

In this way, by integrating the gene amplification part, the electrophoresis part, and the analysis reading part in the miniaturized and portable DNA analysis all-in-one chip device 1, samples can be automatically injected into the electrophoresis pad after PCR is performed and PCR separation is performed. Since it can be carried out at the same time, it is possible to prevent the hassle of moving the PCR amplification material or the contamination during the transfer process. In addition, since the time required from gene amplification to analysis reading is greatly reduced, it is relatively convenient at companies and research institutes and institutes and local governments that inspect agricultural and livestock products because of the low cost and on-site contamination prevention and rapid diffusion. Immediate inspection is possible.

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

1: Integral chip device for DNA analysis
10: top plate 11: air inlet
12: sample inlet 13: through hole
14: gel solution injection port 15: DNA collection port
20: DNA film chip
21: air injection section 22; Sample injection part
23: gene amplification part 24: gel solution injection part
25: DNA collection section 26: electrophoresis section
30 Bottom
40: fixed coupling band

Claims (5)

It is seated on the sample deck and fixed,
Integral chip device for DNA analysis, characterized in that it comprises a gene detection and analysis module that is amplified and electrophoretic by the introduction of a DNA sample.
According to claim 1, The gene detection analysis module,
Top plate 10;
A DNA film chip 20 disposed to correspond to a lower portion of the top plate 10; And
An integral chip device for DNA analysis, characterized in that it is fixedly fastened to the lower half of the upper plate, and is composed of a lower plate (30) with a rear end of the DNA film chip (20) interposed between the upper plate (10).
According to claim 2,
The top plate 10,
From the upper end, the air injection port 11, the sample injection port 12, the through hole 3, the gel solution injection port 14 and the DNA collection port 15 are sequentially formed in a plate shape,
The DNA film chip 20,
The electrophoresis section 26 including the air injection section 21 and the sample injection section 22, the gene amplification section 23 and the gel solution injection section 24 and the DNA collection section 25 is formed of fine capillaries,
The lower plate 30,
Characterized in that the contact point to which electricity can be applied is provided at the upper and lower ends to separate the nucleic acid of the amplified sample from the electrophoretic section 26, and to form an analysis reading section together with the upper panel 10. All-in-one chip device for DNA analysis.
According to claim 2,
The rear end portion of the DNA film chip (25) is an integrated chip device for DNA analysis, characterized in that fixed between the upper plate (20) and the lower plate (27) and fixed with a fixing band (40).
The method according to any one of claims 1 to 4,
The DNA analysis chip device,
A chip device for DNA analysis that is configured to continuously perform gene amplification, electrophoresis, and analysis reading, thereby reducing amplification time, electrophoretic time, and time to analysis reading.

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