KR102157380B1 - Integrated Chip for DNA Analyzer - Google Patents

Abstract

The present invention relates to a portable DNA analysis chip apparatus, comprising: a top plate 10; A DNA film chip 20 disposed to correspond to the lower portion of the upper plate 10; And a lower plate 30 fixedly coupled to the lower half of the upper plate and interposed between the rear end of the DNA film chip 20 between the upper plate 10 and the upper plate 10. Through this, a series of processes of extracting and amplifying DNA genetic material from a sample, separating a number of specific genes at once using a capillary electrophoresis device, and reading the results from the separated images can be performed quickly in the field. It is configured to be able to continuously perform amplification, electrophoresis, and analysis reading, and has the effect of reducing the time required for amplification, time required for electrophoresis, and time until analysis reading.

Description

Integrated Chip for DNA Analyzer {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 easily and conveniently perform the entire process on one chip 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, but in order to proceed with such a PCR method, a thermal cycler device and collection that can amplify the collected DNA 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, is configured to operate in connection with an external computer for data processing, and the internal structure is designed to have low spatial efficiency. Therefore, it can be said that there are many inefficient and unnecessary parts to be used in the actual field.

In particular, if you want to analyze the DNA of an animal suspected of foot-and-mouth disease in order to determine a hereditary infectious disease such as foot-and-mouth disease, or because this DNA must be analyzed by moving it to a place where a real-time PCR equipment is installed, it will leave the area and secondary infection will occur Not only is there a concern, but also a serious problem that is difficult to manage quarantine according to travel time can occur.

In addition, molecular biology tests, which can be diagnosed early by confirming the presence or absence of direct factors such as genetic material of an infectious agent, have a risk of deriving false positive results by the tester or contamination.

Accordingly, in recent years, WHO has emphasized the necessity of on-site inspection for diagnosing infectious diseases, and systems for on-site inspection using molecular biological methods have been steadily developed from about 10 years ago.

Moreover, with the advent of lab-on-a-chip (LOC) technology that allows all molecular diagnosis procedures including extraction of genetic material from a specimen, gene amplification reaction, and diagnosis of amplification, etc. Research progress on possible high-efficiency and high-sensitivity systems is being conducted more actively.

As a result of such research, a strip sensor module for a molecular diagnostic field test device disclosed in Korean Patent Registration No. 10-1767978, and a real-time PCR and gene analysis DNA microarray for quantitative detection of genes disclosed in Korean Patent No. 10-1302353 There are automatic molecular diagnostics analysis devices that can be performed in a complex manner.

However, in the case of such a conventional molecular diagnostic field test device, in general, the configuration of the DNA chip and the measurement device is complicated, so the product manufacturing unit cost is high, and the cost of using a single test is high. Since it has to be transferred to a separate measuring device, there is a disadvantage of lowering the measurement reliability due to contamination during the transfer process, and since gene extraction, amplification, separation, and reading are not performed within one system, initial determination is possible to some extent, but the final result There is a disadvantage that it takes a lot of time to obtain and is not suitable for field inspection.

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

The present invention was conceived to solve the above-described problem, and an object of the present invention is to amplify the DNA genetic material extracted from a specimen using an integrated single chip device, and use a capillary electrophoresis device to It is to provide an all-in-one chip device for DNA analysis configured to perform a series of processes of separating at once and reading the results from the separated images in real time quickly in the field.

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

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

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

The gene detection and analysis module,

Upper plate 10;

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

It is fixedly coupled to the lower half of the upper plate, characterized in that it is composed of a lower plate 30 interposed between the rear end of the DNA film chip 20 and 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 inlet 11, the sample inlet 12, the through hole 3, the gel solution inlet 14 and the DNA collection port 15 are sequentially formed in a plate shape,

The DNA film chip 20,

An electrophoretic unit 26 including an air injection unit 21 and 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 microcapillary tube,

The lower plate 30,

A contact point to which electricity can be applied is provided at the upper and lower ends to separate the nucleic acid of the sample amplified by the electrophoretic unit 26, and forming an analysis and reading unit together with the upper plate 10. All-in-one chip device for DNA analysis.

do.

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

The rear end of the DNA film chip 20 is coupled between the upper plate 10 and the lower plate 30 and is fixed with a fixed binding strip 40.

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

The integrated chip device for DNA analysis,

It is configured to be able to continuously perform gene amplification, electrophoresis, and analysis reading, and is characterized in that it reduces the time required for amplification, the time required for electrophoresis, and the time until analysis reading.

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

In addition, the integrated chip device for DNA analysis according to the present invention is configured to be able to continuously perform gene amplification, electrophoresis and analysis reading, and has the effect of reducing the time required for amplification, time required for electrophoresis, and time until analysis reading. have.

1 is a perspective view briefly showing the internal structure of the 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 of the present invention will be described in more detail with reference to the accompanying drawings.

1 is a perspective view schematically showing the 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 and 2, the integrated chip device 1 for DNA analysis according to the present invention amplifies a DNA sample extracted from the collected sample for a desired DNA region, and converts the amplified sample into a gel solution. It is an integrated chip that moves and separates according to the voltage applied within, and continuously processes the process of photographing, analyzing, and reading the separated sample image, consisting of a gene detection and analysis means mounted on a sample deck (not shown). do.

One side of the sample deck (not shown) is fixed to the inside of the front part of the case of the miniaturized DNA analysis device to enable on-site inspection and is connected to a connected sliding motor to slide in the front and rear direction.The gene detection and analysis module is mounted on the upper surface. A rectangular groove having a predetermined width and length is formed.

The gene detection and analysis means is amplified and electrophoresed by inputting the extracted DNA sample, and is composed of an upper plate 10, a DNA film chip 20 and a lower plate 30.

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

The DNA film chip 20 is a linear electrophoresis unit 26 comprising an air injection unit 21 and 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 microcapillary tube, and 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 bonding strip. The thickness of the film chip is manufactured to be approximately 430, of course, but is not limited thereto.

The air injection unit 21 and the sample injection unit 22 are provided with an air injection hole and a sample injection hole communicating with the air injection port 11 and the sample injection port 12 of the upper plate 10, respectively, so that the supplied air is a DNA film. The remaining air bubbles and excess sample contained in the chip can be discharged, and the DNA sample extracted from the sample can be added.

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

The gene amplification unit 23 amplifies a plurality of extracted DNA for each 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 electrophoretic unit 26 is formed by injection molding of a plurality of microchip-based microcapillaries, and is formed to be continuous with the gene amplification unit 23.

The gel solution is injected into the injection hole of the gel solution injection unit 24 communicated 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 reference marker formed on the side of the sample containing the DNA to be measured and a capillary tube of the same size provided with the gel solution, so that the sample is the same as the amplified sample of the sample. Allows separation under conditions.

In this way, by providing the reference marker on the side and separating it with the specimen at the same time, analysis differences due to external environmental factors and condition variations in the separation process do not occur, enabling more precise analysis and reading.

Here, polyacrylamide gel or agarose gel may be used as a gel solution as a support, and it goes without saying that the gel has functions such as convection prevention, sample protection, and molecular sieve.

In addition, it is of course that it is possible to collect the nucleic acid separated from the DNA collection unit 25.

The lower plate 30 is in the shape of a transparent rectangular plate having a predetermined width and length, and has contacts at the upper and lower ends to which electricity can be applied to separate the nucleic acids of the sample amplified by the electrophoretic unit 26 by the applied electricity. It enables comparative measurement, fixes the film chip interposed between the upper plate and the upper plate, and compares a plurality of linear images separated by the linear electrophoresis unit 26 with a reference marker to form an analysis reading unit that analyzes and reads. .

The size from the front end to the lower end of the lower plate 30 has a length corresponding to the 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 produced, of course, but is not limited thereto.

The fixed coupling strip 40 is fixedly coupled to the rear portion of the upper plate and the lower plate and the film chip interposed therebetween on the upper plate 10, and a coupling protrusion is 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 is connected to the gel solution injection port 24 to prevent the solution from leaking to the outside during the process.

The common region of the lower plate corresponding to the upper plate is an analysis and reading unit, and analysis and reading are performed by comparing a plurality of linear images from which nucleic acids are separated by the electrophoresis unit 26 with a reference marker. For this analysis, an imaging device for photographing an analysis and reading unit may be provided on the upper side and a light guide plate serving as a lower illumination may be provided at the lower part.

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

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

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

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

Then, the prepared DNA sample is injected into the capillary tube of the sample injection unit through the sample injection port through the sample injection unit (not shown) or through the use of a dispenser or by activating the injection operation button.

When the injected DNA sample is quickly moved to the gene amplification unit due to a capillary phenomenon, amplification is performed by heating and cooling the gene amplification unit through a heating and cooling device.

The amplified sample is injected into the capillary of the electrophoretic unit 26 connected to communicate with each capillary tube, and the gel solution is injected through the gel solution injection port.

Next, when a DC voltage is applied to the electrophoresis unit, charged particles move from 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 the moving speed varies according to the molecular weight. At this time, the gel solution and the gene move rapidly forward due to the capillary phenomenon according to the microchip-based capillary structure, and the movement speed is fast, and accordingly, the analysis can be performed in a short time.

Next, a plurality of linear images measured separated from the electrophoresis unit are photographed using an imaging device and LED light, compared with a reference marker through an internal analysis program, and analyzed and read, and the analyzed result value is obtained from a display device or It may be provided so as to appear on a separate play means.

In this way, by integrating the gene amplification unit, the electrophoresis unit, and the analysis reading unit in the miniaturized and portable DNA analysis unit 1, the sample can be automatically injected into the electrophoresis pad after PCR is performed, and PCR separation can be performed. Since it can be performed at the same time, it is possible to prevent the hassle of moving the PCR amplification material or contamination during the transfer process. In addition, since the time required from gene amplification to analysis and reading is greatly reduced, it is relatively simple and convenient in companies, research institutes, and local governments that are autopsies of agricultural and livestock products due to the time-critical contamination sites or sites to prevent diffusion due to rapid spread and low prices. Immediate inspection becomes possible.

Although the present invention has been described with reference to embodiments with reference to the drawings, this is only exemplary, and those of ordinary skill in the art will understand that various modifications and equivalent embodiments are possible therefrom. Accordingly, the true technical scope of the present invention should be determined by the technical spirit of the appended claims.

1: All-in-one chip device for DNA analysis
10: top plate 11: air inlet
12: sample injection port 13: through hole
14: gel solution injection port 15: DNA collection port
20: DNA film chip
21: air injection unit 22; Sample injection part
23: gene amplification unit 24: gel solution injection unit
25: DNA collection unit 26: electrophoresis unit
30 lower plate
40: fixed binding strip

Claims (5)

It is seated and fixed on the sample deck,
It comprises a gene detection and analysis module for amplification and electrophoresis by inserting a DNA sample,
The gene detection and analysis module,
Upper plate 10; A DNA film chip 20 disposed to correspond to the lower portion of the upper plate 10; And a lower plate 30 which is fixedly fastened with the lower half of the upper plate and interposed with the rear end of the DNA film chip 20 between the upper plate 10,
The top plate 10,
It is a plate shape in which an air inlet 11, a sample inlet 12, a through hole 3, a gel solution inlet 14 and a DNA collection port 15 are sequentially formed from the upper end,
The DNA film chip 20,
The electrophoretic unit 26 including the air injection unit 21 and the sample injection unit 22, the gene amplification unit 23 and the gel solution injection unit 24 and the DNA collection unit 25 is formed of a microcapillary tube,
The lower plate 30,
A contact point to which electricity can be applied is provided at the upper end and the lower end to separate the nucleic acid of the sample amplified by the electrophoretic unit 26, and to form an analysis reading unit together with the upper plate 10. All-in-one chip device for DNA analysis.
delete delete The method of claim 1,
An integrated chip device for DNA analysis, characterized in that the rear end of the DNA film chip (20) is coupled between the upper plate (10) and the lower plate (30) and fixed with a fixed binding strip (40).
The method of claim 1 or 4,
The integrated chip device for DNA analysis,
An integrated chip device for DNA analysis, characterized in that it is configured to be able to continuously perform gene amplification, electrophoresis, and analysis reading, thereby reducing the time required for amplification, time required for electrophoresis, and time until analysis reading.

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