KR100921752B1 - Apparatus for analyzing DNA fingerprint image and method therefor - Google Patents

Abstract

The present invention relates to a genetic fingerprint image analysis apparatus and a method thereof, and more particularly, to a genetic fingerprint image analysis apparatus and method for detecting and analyzing information of genes using computer image processing technology.

The present invention provides a device for analyzing a genetic fingerprint image, comprising: an extractor extracting DNA by amplifying gene information of a DNA sample; An ultraviolet irradiation unit for irradiating ultraviolet rays with DNA extracted through the extraction unit; An image acquisition unit for obtaining a gene fingerprint image from DNA irradiated with ultraviolet rays through an ultraviolet irradiation unit; A spectrophotometer for identifying DNA quantification from genetic fingerprint images; An image analyzer for correcting the skewing phenomenon and binarizing the genetic fingerprint image; An output unit for outputting an analysis result of the image analyzer; And a control unit which controls an extraction unit, an ultraviolet irradiation unit, an image acquisition unit, a spectrophotometer, an image analyzer, and an output unit. It includes.

According to the present invention, there is an effect of obtaining a genetic fingerprint image by using computer image processing technology, and correcting the distortion phenomenon generated during electrophoresis for the recognition of the genetic fingerprint image.

Genetic Fingerprint Imaging, Distortion, Correction, Binarization

Description

Apparatus for analyzing DNA fingerprint image and method therefor}

The present invention relates to a genetic fingerprint image analysis apparatus and a method thereof, and more particularly, to a genetic fingerprint image analysis apparatus and method for detecting and analyzing information of genes using computer image processing technology.

Detecting the information of genes in gene fingerprint images is of considerable value in and of itself, and analyzing and processing this data can yield more biological information needed by biotechnology.

Conventionally, a technique for analyzing a genetic fingerprint image has been published and published in a number of applications other than the Republic of Korea Patent Publication No. 10-2002-0079035 'method for examining the genetic fingerprint'.

The genetic fingerprinting method may include extracting DNA from the ear canal secretion; Performing PCR on the DNA sample; And separating the amplified DNA fragments.

Research into the molecules that make up the body of life, such as DNA, RNA, and proteins, is proceeding more and more actively due to its importance, and the structures of molecules that have not been revealed until now are rapidly revealed through experiments. The structural data of molecules revealed through this experiment are being systematically constructed in database, and more and more molecular structure data will be databased in the future.

However, until now, the method of acquiring the information of the genetic fingerprint information has been manually obtained and stored directly on the computer. This is a problem that is not uniform, depending on the professionalism of the subject to obtain genetic fingerprint information and changes in the individual information acquisition environment.

Disclosure of Invention An object of the present invention is to provide a genetic fingerprint image analysis device and method for acquiring a genetic fingerprint image using computer image processing technology.

Another object of the present invention is to provide an apparatus and method for analyzing a genetic fingerprint image for correcting distortion occurring during electrophoresis for recognition of a genetic fingerprint image.

Another object of the present invention is to provide a genetic fingerprint image analysis apparatus and method for binarizing each lane for recognition of a genetic fingerprint image irrelevant to a smear phenomenon.

An apparatus for analyzing a genetic fingerprint image, the apparatus comprising: an extraction unit configured to extract DNA by amplifying gene information of a DNA sample; An ultraviolet irradiation unit for irradiating ultraviolet rays with DNA extracted through the extraction unit; An image acquisition unit for obtaining a gene fingerprint image from DNA irradiated with ultraviolet rays through the ultraviolet irradiation unit; A spectrophotometer for identifying DNA quantification from the genetic fingerprint image; An image analyzer for analyzing the genetic fingerprint image to correct tilting and binarization; An output unit for outputting an analysis result of the image analyzer; And a control unit controlling the extraction unit, the ultraviolet irradiation unit, the image acquisition unit, the spectrophotometer, the image analyzer and the output unit. It includes.

On the other hand, in the genetic fingerprint image analysis method, (a) a step of extracting DNA by amplifying the genetic information of the DNA sample by the control unit of the genetic fingerprint image analysis unit; (b) electrophoresis of the controller by the extracted DNA; (c) the control unit irradiating ultraviolet rays with DNA extracted from the extraction unit through an ultraviolet irradiation unit; (d) acquiring, by the control unit, a genetic fingerprint image from the DNA irradiated with ultraviolet rays through the image acquisition unit; (e) checking, by the controller, whether DNA is present in the genetic fingerprint image; (f) the control unit confirming DNA quantification through a spectrophotometer; (g) correcting, by the controller, a tilting phenomenon of the genetic fingerprint image through an image analyzer; And (h) binarizing the gene fingerprint image for each lane by the image analyzer; It includes.

The present invention has the effect of obtaining a genetic fingerprint image using computer image processing technology.

In addition, there is an effect of correcting the distortion occurring during electrophoresis for the recognition of the genetic fingerprint image.

In addition, there is an effect to binarize for each lane (lane) for the recognition of the genetic fingerprint image irrelevant to the smear phenomenon.

In addition, there is an effect to solve the inaccuracies caused by the researcher's manual and visual marker recognition, and reduce manpower, time and cost.

The features and advantages of the present invention will become more apparent from the following detailed description based on the accompanying drawings. Prior to this, the terms or words used in the present specification and claims are defined in the technical spirit of the present invention on the basis of the principle that the inventor can appropriately define the concept of the term in order to explain his invention in the best way. It should be interpreted to mean meanings and concepts. In addition, when it is determined that the detailed description of the known function and its configuration related to the present invention may unnecessarily obscure the subject matter of the present invention, it should be noted that the detailed description is omitted.

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

The apparatus for analyzing a genetic fingerprint image according to an embodiment of the present invention will be described with reference to FIGS. 1 to 9.

1 is a block diagram of an apparatus for analyzing a gene fingerprint image according to an embodiment of the present invention, FIG. 2 is a phenomenon in which the lane of the gene fingerprint image is inclined according to an embodiment of the present invention, and FIG. Figure 4 is a smear phenomenon of a gene fingerprint image according to an embodiment of the present invention, Figure 4 is a phenomenon of severe severity of the genetic fingerprint image according to an embodiment of the present invention, Figure 5 is a view of the lane according to an embodiment of the present invention Before and after the slope correction, Figure 6 is a histogram of the binarized image according to an embodiment of the present invention, Figure 7 is a vertically projected image diagram according to an embodiment of the present invention, Figure 8 is the present invention FIG. 9 is a lightness distribution diagram of the entire image according to an embodiment of the present invention, and FIG. 9 is a lightness distribution diagram of the image for lane 6 according to an embodiment of the present invention.

Gene fingerprint image analysis apparatus according to an embodiment of the present invention, as shown in Figure 1, the extraction unit 100, ultraviolet irradiation unit 200, image acquisition unit 300, spectrophotometer 400, image analysis The unit 500 includes an output unit 600 and a control unit 700.

Extraction unit 100 performs PCR (Polymerase Chain Reaction) on DNA samples extracted from body organs or individuals to amplify and extract DNA with a large amount of genetic information.

The PCR is to amplify a desired DNA molecule in vitro by specifically repeatedly synthesizing a specific DNA region, and amplified into DNA having a large amount of genetic information using a very small amount of DNA. Generally, the PCR reaction mixture contains sample DNA as a template, two primers, 'tag DNA polymerase', and four 'dNTP'. As the double primer, oligonucleotides synthesized to bind complementarily to the three-terminal sequence of each single helix in a specific DNA fragment are used. First, PCR is timed by applying high temperature to the reaction mixture to separate the template DNA into single helix. Subsequently, by lowering the temperature, the primers in the mixed solution bind to each single helix of the target DNA, and a new DNA strand is synthesized by 'tag DNA polymerase'. Thus, new DNA synthesis proceeds along the DNA sequence between the two primers. Thus, by one cycle of PCR, the specific DNA fragments are doubled and they act as target DNA of the next PCR. After a continuous n cycles, the number of specific DNA fragments is exponentially amplified to 2n. will be.

In addition, the ultraviolet irradiation unit 200 serves to irradiate ultraviolet rays with DNA having a large amount of genetic information extracted through the extraction unit 100.

In addition, the image acquisition unit 300 obtains a genetic fingerprint image from the DNA irradiated with ultraviolet rays by the ultraviolet irradiation unit 200. The image acquisition unit 300 according to the present embodiment acquires at least one or more gene fingerprint images of RAPD, RFLP, AFLP, SSR, and CAPs.

In addition, the spectrophotometer 400 checks the DNA quantification in the gene fingerprint image obtained through the image acquisition unit 300.

The spectrophotometer 400 will be described below. When a molecule absorbs ultraviolet light, it causes the binding electrons and non-binding electrons to transition from the ground state to the excited state and then release energy by returning to the ground state through several paths. do. Spectrophotometer 400 observes these changes to determine the electronic structural properties of the molecule.

DNA quantification of the spectrophotometer 400 according to the present embodiment will be described with reference to the protein as follows. Tyrosine, phenylalanine and tryptophan residues in the protein absorb ultraviolet light at 275 nm and 280 nm, respectively. For proteins, the combined level of amino acids is nearly constant so the concentration of the protein is proportional to the absorbance at 280 nm. Of course, with the exception of pure proteins, it can be quickly determined by measuring the absorbance at 280 nm when the protein concentration is 1 mg / ml and the path of ultraviolet light is 1 cm, which means less time to quantify There is an advantage in that the protein can be recovered and reused after confirmation.

In addition, the image analyzer 500 corrects the tilt of the image by analyzing the genetic fingerprint image acquired through the image acquisition unit 300 and binarizes each lane. In order to perform this function, the image analyzer 500 includes a calibration module 510 and a binarization module 520.

Genetic fingerprint image according to an embodiment of the present invention has the following characteristics. As shown in FIG. 2 due to the foreign matter on the gel during electrophoresis, there is a phenomenon that the lane is inclined. In addition, due to the difference in voltage and gene concentration during electrophoresis, as shown in FIG. 3, there is a distortion phenomenon of marker information in which a bright band in a vertical direction occurs. This is called smearing. In addition, as shown in FIG. 4 due to the foreign matter on the gel during electrophoresis, there is a blemish in the image.

As described above, the calibration module 510 corrects a phenomenon that the lane is inclined due to foreign matter on the gel during electrophoresis.

The calibration module 510 according to the present embodiment will be described below.

,

) 매개 변수 평면의 곡선으로 Hough 변환시킨다. The calibration module 510 tracks the contour of the genetic fingerprint image, and uses one of the following equations (x, y) of the inclined portion of the (X, Y) plane to obtain the following equation (1):

,

) Convert the Hough to a curve in the parameter plane.

는 원점으로부터 (x, y)를 지나는 임의의 직선까지의 거리이며,

는 그 직선의 법선 벡터가 X축과 이루는 각도이다. here,

Is the distance from the origin to any straight line through (x, y),

Is the angle that the normal vector of the straight line makes with the X axis.

,

에 따라 곡선들이 교차하는 수를 누적한다. 이 누적값을 임계치와 비교하여 선 성분인지를 판단할 수 있으며, 검출된 선 성분의 기울기는 법선 벡터의 기울기를 이용하여 계산할 수 있다.The Hough transform draws a trajectory on the parameter plane for each black pixel of the input image.

,

Accumulate the number of intersections of the curves. The cumulative value may be compared with a threshold to determine whether it is a line component, and the detected slope of the line component may be calculated using the slope of the normal vector.

Gradient correction uses a shearing equation, such as the following equation 2, which is simple and relatively accurate to perform in lanes.

는 추정된 기울기이고

는 입력 숫자의 기본 기울기를 나타낸다. 본 실시예에 따른 유전자 지문영상에 대한 기울기 교정 전(a), 후(b)는 도 5에 도시된 바와 같다. Where (x, y) is a point of the tilted portion of the image and (x ', y') represents a point of the converted image.

Is the estimated slope

Denotes the default slope of the input number. Before (a) and after (b) the slope correction for the genetic fingerprint image according to the present embodiment is as shown in FIG.

In addition, the binarization module 520 functions to binarize the genetic fingerprint image in order to solve the smear miscellaneous.

Referring to the binarization module 520 according to the present embodiment.

Binarization is a technique of expressing an image with only two values, black and white. When expressed as a brightness value, binarization has a value of 0 or 255. A more general expression is that it only has a value of 0 or 1. In this case, a value used as a reference for dividing an image by a value of 0 or 1 is called a threshold. That is, binarization in a gray image is a process of determining a certain threshold value and making the brightness value 0 (black) if the brightness value of the pixel is smaller than the threshold value and making the brightness value 255 (white) if it is larger than the threshold value.

When an image has a unique brightness in the background and the foreground, and the brightness difference between them is sufficiently distinguished, since the histogram of the binarized image has two distribution shapes as shown in FIG. 6, the threshold is determined by finding a valley on the histogram. Can be easily binarized. However, although there are two peaks on the histogram, they do not always point to the foreground and background in the actual image, and sometimes only one peak appears but the foreground and background are clearly distinguished in the actual image. Therefore, a histogram that has only statistical characteristics of brightness values may not reflect the state of the image well.

These binarization methods can be divided into global binarization methods and local binarization methods. When global binarization is performed, when an outlier appears in an intermediate region of an image, it is impossible to provide accurate information about the corresponding region. To overcome this, binarization by dividing into meaningful local space is not affected by outliers.

비율만큼 삭제(

-cut)시킨다. 상기

비율은 잡영에 의해 누적된 레인영상 간에 연결된 임의의 값이다.The binarization module 520 divides the vertically projected genetic fingerprint image into regions for each lane. Genetic fingerprint images consisting of eight lanes are affected by various conditions of electrophoresis, so if the lanes are recognized as one image connected to each other, marker recognition by simple binarization will fail. none. Therefore, as shown in FIG. 7, the binarization module 520 divides lane regions when the parts connected to each lane are completely divided by 0 when vertical projection is performed, as shown in FIG. 7. When a connection is made between lane images accumulated by

Delete by percentage

-cut) remind

The ratio is an arbitrary value connected between lane images accumulated by ghosting.

In addition, the output unit 600 performs a function of outputting the result analyzed by the image analyzer 500.

The control unit 700 controls the extraction unit 100, the ultraviolet irradiation unit 200, the image acquisition unit 300, the spectrophotometer 400, the image analyzer 500, and the output unit 600. Electrophoresis to confirm whether the DNA extracted through 100 is normally extracted, the presence or absence of DNA in the genetic fingerprint image obtained through the image acquisition unit 300, and the brightness average and Perform the function of calculating the standard deviation.

When comparing the brightness average and standard deviation of the entire image of the genetic fingerprint image according to the present embodiment, and the brightness average and standard deviation of the image localized by each lane, it is shown in Table 1 above.

FIG. 8 is a brightness distribution map of the entire image, and FIG. 9 is a brightness distribution map of the image for lane 6. FIG.

Referring to Tables 1, 8 and 9, it can be seen that the brightness average and standard deviation of the local image are better representative values than the brightness average and standard deviation of the entire image.

Meanwhile, the overall flow of the method (hereinafter, referred to as 'genetic fingerprint image analysis method') using the gene fingerprint image analysis device according to an embodiment of the present invention will be described with reference to FIG. 10.

10 is an overall flowchart of a genetic fingerprint image analysis method according to an embodiment of the present invention.

As shown in FIG. 10, the control unit 700 performs a polymerase chain reaction on a DNA sample extracted from a body organ or an individual through the extracting unit 100 to extract DNA having a large amount of genetic information (S2). ).

Next, the control unit 700 is electrophoresed with DNA having a large amount of genetic information to confirm whether the DNA is normally extracted (S4).

Next, the control unit 700 irradiates the ultraviolet light with DNA extracted from the extraction unit 100 through the ultraviolet irradiation unit 200 (S6).

Next, the control unit 700 obtains a genetic fingerprint image from the DNA irradiated with ultraviolet rays through the image acquisition unit 300 (S8).

Next, the controller 700 checks the presence or absence of DNA in the obtained genetic fingerprint image (S10).

If the DNA is not confirmed as a result of step S10, the procedure is performed before step S2.

Next, the control unit 700 confirms DNA quantification through the spectrophotometer 400 (S12).

Next, the controller 700 analyzes the gene fingerprint image through the image analyzer 500 and corrects the skewing phenomenon (S14).

Next, the controller 700 binarizes the gene fingerprint image for each lane through the image analyzer 500 (S16).

The controller 700 outputs the result analyzed by the image analyzer 500 through the output unit 600 (S18).

A detailed flow of the tilt phenomenon correction step according to an embodiment of the present invention will be described with reference to FIG. 11 as follows.

11 is a detailed flowchart of the tilt phenomenon correction step according to an embodiment of the present invention.

As shown in FIG. 11, the controller 700 tracks the contour of the genetic fingerprint image through the correction module 510 of the image analyzer 500 (S22).

,

) 평면의 곡선으로 Hough 변환시킨다(24).Next, the control unit 700 sets a parameter (x, y) of the inclined portion of the (x, y) plane as a parameter (

,

Hough transform into a curve of the plane (24).

In addition, the controller 700 corrects the tilting phenomenon by converting the shearing through the calibration module 510 (S26).

A detailed flow of each lane binarization step according to an embodiment of the present invention will be described with reference to FIG. 12.

12 is a detailed flowchart of the binarization step for each lane according to an embodiment of the present invention.

As shown in FIG. 12, the controller 700 vertically projects the genetic fingerprint image through the binarization module 520 of the image analyzer 500 (S32).

Next, the control unit 700 divides the vertically projected genetic fingerprint image by each lane through the binarization module 520 (S34).

The control unit 700 calculates the brightness average and the standard deviation for each of the entire image and the divided lane image through the binarization module 520 (S36).

As described above and described with reference to a preferred embodiment for illustrating the technical idea of the present invention, the present invention is not limited to the configuration and operation as shown and described as described above, it is a deviation from the scope of the technical idea It will be understood by those skilled in the art that many modifications and variations can be made to the invention without departing from the scope of the invention. Accordingly, all such suitable changes and modifications and equivalents should be considered to be within the scope of the present invention.

1 is a block diagram of a genetic fingerprint image analysis apparatus according to an embodiment of the present invention.

2 is a phenomenon in which the lane of the genetic fingerprint image is inclined in accordance with an embodiment of the present invention.

Figure 3 is a smear phenomenon of the genetic fingerprint image according to an embodiment of the present invention.

Figure 4 is a severe phenomenon of the genetic fingerprint image according to an embodiment of the present invention.

5 is a view showing before and after the slope correction of the lane according to an embodiment of the present invention.

6 is a histogram of a binarized image according to an embodiment of the present invention.

7 is a vertical projection image in accordance with an embodiment of the present invention.

8 is a brightness distribution diagram of an entire image according to an embodiment of the present invention.

9 is a brightness distribution diagram of an image for lane 6 according to an embodiment of the present invention.

Figure 10 is a flow chart of the genetic fingerprint image analysis method according to an embodiment of the present invention.

11 is a detailed flowchart of the tilt phenomenon correction step according to an embodiment of the present invention.

12 is a detailed flowchart of the binarization step for each lane according to an embodiment of the present invention.

<Description of Drawing>

100: extraction unit 200: ultraviolet irradiation unit

300: image acquisition unit 400: spectrophotometer

500: image analyzer 600: output unit

700: control unit

Claims (7)

In the genetic fingerprint image analysis device, Extracting unit 100 for amplifying the DNA information of the DNA sample to extract DNA; An ultraviolet irradiation unit 200 for irradiating ultraviolet rays with DNA extracted through the extraction unit; An image acquisition unit 300 for obtaining a gene fingerprint image from DNA irradiated with ultraviolet rays through the ultraviolet irradiation unit; A spectrophotometer 400 for confirming DNA quantification from the genetic fingerprint image; The contour of the genetic fingerprint image is traced to determine a point (x, y) of the inclined portion of the (x, y) plane as a parameter (

,

Hough transformation to flat curve, shearing transformation to correct tilting, vertical projection of the genetic fingerprint image, and segmentation of the vertically projected genetic fingerprint image for each lane. An image analyzer 500 for deleting and binarizing by a random value α connected between the accumulated lane images by the ratio; An output unit 600 for outputting an analysis result of the image analyzer; And A controller 700 for controlling the extraction unit, the ultraviolet irradiation unit, the image acquisition unit, the spectrophotometer, the image analyzer and the output unit; Genetic fingerprint image analysis device comprising a. The method of claim 1, The extraction unit 100, Gene fingerprint image analysis device, characterized in that for amplifying DNA molecules using a polymerase chain reaction to the DNA sample. The method of claim 1, The image acquisition unit 300, Gene fingerprint image analysis device, characterized in that to obtain at least one or more genetic fingerprint image of RAPD, RFLP, AFLP, SSR and CAPs. The method of claim 1, The image analyzer 500, The contour of the genetic fingerprint image is traced to determine a point (x, y) of the inclined portion of the (x, y) plane as a parameter (

,

A correction module 510 for correcting a tilting phenomenon by converting the Hough into a curve of a plane and then performing a shearing conversion; And A binarization module for vertically projecting the gene fingerprint image and deleting and binarizing the vertically projected gene fingerprint image by an arbitrary value (α) ratio connected between lane images accumulated by ghosting in order to segment regions by each lane ( 520); Gene fingerprint image analysis device comprising a. In the genetic fingerprint image analysis method, (a) amplifying gene information of a DNA sample by the control unit 700 of the gene fingerprint image analyzing apparatus through the extraction unit 100 to extract DNA; (b) electrophoresis of the controller by the extracted DNA; (c) the control unit irradiating ultraviolet rays with DNA extracted from the extraction unit through the ultraviolet irradiation unit 200; (d) acquiring, by the control unit, a genetic fingerprint image from the DNA irradiated with ultraviolet rays through the image acquisition unit 300; (e) checking, by the controller, whether DNA is present in the genetic fingerprint image; (f) checking, by the controller, DNA quantification through a spectrophotometer (400); (g) The controller tracks the contour of the genetic fingerprint image through the image analyzer 500 to determine a point (x, y) of the inclined portion of the (x, y) plane as a parameter (

,

Hough transform into a curve of the plane, and then shearing to correct the tilt phenomenon; And (h) an arbitrary value connected between lane images accumulated by miscellaneous images in order for the controller to vertically project the genetic fingerprint image through the image analyzer and divide the region of the vertically projected genetic fingerprint image for each lane; α) binarizing by deleting the ratio; Gene fingerprint image analysis method comprising a. The method of claim 5, wherein The step (g) is (g-1) tracking, by the controller 700, the contour of the genetic fingerprint image through the correction module 510 of the image analyzer 500; (g-2) The control unit may determine a point (x, y) of the inclined portion of the (x, y) plane of the gene fingerprint image through the calibration module.

,

Hough transform to a curve of the plane; And (g-3) correcting the tilting phenomenon by the conversion of the controller through the calibration module; Gene fingerprint image analysis method comprising a. The method of claim 5, wherein The (h) step, (h-1) vertically projecting the genetic fingerprint image through the binarization module 520 of the image analyzer 500 by the controller 700; (h-2) dividing an area by each control unit by each lane through the binarization module; And (h-3) calculating, by the controller, brightness averages and standard deviations for the entire image and the divided lane images through the binarization module; Gene fingerprint image analysis method comprising a.

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