TW569010B - Bioagent automatic detection system - Google Patents

Abstract

A bioagent automatic detection system comprises a detection machine and a data processing device. The detection machine is installed with a rail for a bioagent moving in-and-out of the housing space of the detection machine. The housing space is illuminated by two light sources installed symmetrically and in tilt. A camera is used to take the image of a bioagent. The image is transmitted to a data processing device for image treatment. The invented system calculates the image photo intensity in the photo field measurement region between a measurement line and a control line of the bioagent in order to obtain an average gray value as a graystand, and divides the image of the measurement line into n regions. Next, the system compares the graystandard with the control line portion and the image photo field intensity threshold Tn of each region of the measurement line to determine the detection result.

Description

569010 V. Description of the invention (1) [Technical field to which the invention belongs] The present invention relates to an automatic detection system for biological reagents, which has fast, high sensitivity and other concerns. The detection system uses the special arrangement of the backlight panel, CCD, magnifying head and computer to identify the detection results and the results of the reaction. [Previous technology] According to the press, hepatitis C is an inflammation of the liver caused by Hepatitis c virus (HCV). Hepatitis C is transmitted through HCV-infected blood and body fluids through the skin or mucous membranes, and finally into the blood. H C V infection can cause sexual hepatitis. Such chronic hepatitis is usually asymptomatic and is usually found on a regular health checkup. Hepatitis C can cause severe liver complications. Patients often die from severe cirrhosis or hepatocellular carcinoma after 15-20 years of infection. After being infected with HCV, patients will produce anti-HCV antibodies to fight against the invading H C V, and the anti-H C V antibody is the main target of the biological reagent automatic detection system. The reagents used in the biological reagent automatic detection system apply immunoassay and inspection techniques and chromatography principles to fix the recombinant protein of HCV (NS 3, NS 4, NS 5, core protein) at the position of the test line, and combine with PrOtein g Colloidal gold is used as a reagent and coloring agent, mainly to detect anti-HCV antibodies (IgG, IgM, IgA) in the test body, and to quickly and correctly determine whether human blood is infected with HCV to help individuals and physicians make early diagnosis. Samples are serum or plasma, and can be tested immediately after blood extraction by centrifugation. For temporary storage, it must be stored at 2-8. C. If long-term storage is required, it must be stored at -20 ° C.

Page 6569010 —----- —______ V. Description of the invention (2) Since the sensitivity of the biochemical rapid test reagent is much higher than the traditional test, this difficulty can be overcome, so the rapid sensitivity of the biochemical test is high. . θ $ Μ As shown in Figure 1, it is the indication of a rapid hepatitis test reagent, and the biological reagent 8 has an action zone 8 丄 for dripping into the test: liquid or body fluid, a judgment whether the reagent is effective The control line 8 3 is a test line 8 2 for viewing the results of the biochemical reaction, and the area where the control line = test line 82 is located is referred to as a test area A. If this test is performed on the second grade, the color & will appear on the empty line 8 3, and the biochemical reaction result after the test serum drop f 8 1 will be presented on the test line 8 2, so that the reaction color To determine if the blood is infected with hepatitis. "Because the biochemical reaction uses fluorescence or color arrangement, combination, etc. to judge its result" and to detect the sunburn by visual method, general inspectors are prone to deviations due to subjective judgment during visual inspection. & The machine vision system can be used to identify the arrangement of colors and the inspection results. With the development of biochemical technology and the birth of biological crystals and various types of biochemical sensors, machine vision technology has become a trend for the identification and detection of biochemical reactions. For example, Victor et Morozov et al. 2 0 0 One year, a method of detecting array-type protein wafers using a CCD was proposed. Alberto Delgad0 proposed in 2002 a set of D N A arrangement inspection system for MA daily films. In 2005, a method of detecting DNA using CMOS was proposed by j〇〇η μ yong g S og and others. However, because the test line 8 2 on the biological reagent 8 is usually made by a rolling application method, it does not have a very good uniformity, and the edges often have blurring and uneven concentration. And test line 8 2 must be slanted

Page 7569010 V. Description of the invention (3) Only the light can highlight the detection result, but when the oblique light is irradiated on the test area A of the biological reagent 8, it also causes a gradual change in the detection image. The "field measurement area" must use the light field intensity of the reference light field measurement area as the basis for reference detection. The material of the test strip is to be firmly combined with the specimen, so it causes the surface of rice paper or cloud paper, and the detection image often has the effect of dyeing, which brings noise disturbance. These problems will affect the accuracy of machine vision technology to identify the detection results of biological reagents, and even cause misjudgment. Therefore, how to develop a system that can accurately identify the results of biochemical reactions of biological agents has become the subject of the present invention. [Summary of the Invention] The main object of the present invention is to solve the above-mentioned problems and provide a camera to capture the image of the tested area of the biological reagent, and then cut the image of the test line part of the biological reagent into n areas, so that By comparing the thresholds of each area, it is possible to accurately determine the efficacy of the biological reagent test result. In order to achieve the aforementioned object, the biological reagent automatic detection system of the present invention includes: a testing machine having a containing space therein, and the testing machine having an openable and closable door for biological reagents Entering and exiting the accommodating space 5 and the testing machine has several cooling holes' for air circulation, a slide handle is erected in the accommodating space of the testing machine, and a positioning auxiliary rod is provided at the end of the slide rail. , The biological reagent enters and leaves the accommodating space through the slider, and the positioning aid is provided by the positioning auxiliary rod; a light source group 'which is composed of two light sources', and the two light sources are spaced apart

569010 V. Description of the invention (4) At the measurement area corresponding to the biological reagent above the slider, the two light sources are arranged in a symmetrically inclined form, and the direction in which the two light sources are inclined is perpendicular to the extension direction of the far slide In addition, heat radiating fins are attached to each light source to enhance the cooling effect; a camera is set up between the two light sources of the light source group and corresponds to the test area of the biological reagent; more than one fan, which It is set up in the accommodating space, so as to generate airflow, the heat in the accommodating space is exhausted through several heat dissipation holes; a data processing device, which is connected with the camera and biological reagents obtained by the camera The image is subjected to image processing, and the image light field intensity of the light field measurement area between the test line and the control line of the biological reagent is calculated to obtain an average gray value as a threshold value Graystandard, and the image of the test line is vertically and horizontally divided into η regions, and compare the threshold value Graystandard with the image light field intensity threshold τ n of each area of the control line and the test line, the average gray of the image in any area When the value Grayave is lower than the regional threshold Tη, the region is a dangerous reaction, and the grayscale average value Grayp 'of all positive reaction regions is calculated, and the level of the biochemical reaction concentration is calculated based on the grayscale average. The degree Grayave is higher than the regional threshold, so that the detection result of the biological reagent is determined as a negative reaction. The reagents used in the biological reagent automatic detection system of the present invention apply immunoassay testing technology and chromatography principles to fix HCV recombinant proteins (NS3, NS4, NS5, core protein) at the position of the test line, and the specimen rises to the position of the test line 1 ～ 5 minutes, the color is judged by the visual detection system, and the image is read and interpreted by the computer. The detection result can be known within 0.3 seconds,

569010 V. Description of the invention 高 High sensitivity and very simple operation. The above and other advantages and advantages of the present invention 5 It is not difficult to obtain a deep understanding from the detailed description and drawings of the selected embodiment below. Of course? Some of the parts or arrangements of other parts are allowed to be different, but the selected embodiment is described in detail in this specification, and its structure is shown in the drawings. [Embodiment] Please refer to Figure 1 To the 16th figure, the structure shown in the figure is the selected embodiment of the structure of the present invention. This is for illustration only. In the patent, please do not limit this structure. The biological reagent dynamic detection system of the invention 5 It includes a testing machine 1 part of the hardware and a data processing device part of the software. There is an accommodating space 1 1 j, and the testing machine 1 has a door 12 that can be opened and closed for biological reagents 8 to enter and leave the accommodating space 11, and the testing machine 1 has several cooling holes 1 on both sides of the testing machine 1. 3 for air circulation. 0 * The accommodating space 1 1 of the inspection machine 1 has ri-Π. Δ again. • There is a slide rail 1 4 5 The frame δ is in the accommodating space 1 of the inspection machine 1 1 1 A positioning aid rod 1 4 1 is provided at the end of the slide rail 1 4 for biological reagents 8 Follow the slider 1 4 to enter and leave the accommodation space 1 1 and provide positioning function by the positioning assistance rod 1 4 1 The group is composed of two light sources 1 5 in this embodiment, and the light source is a yellow light backlight plate 1. The two light sources 15 are a spacer ^ rTL δ and correspond to the test area A of the biological reagent 8 above the slide rail 1 4 To be " light

Page 569569010 V. Description of the invention (6) The source 15 is symmetrically inclined, and the direction of the two light sources 15 is perpendicular to the extension direction of the slider 14 and on the light sources 15 Attach heat sink fins 1 5 1 俾 to enhance heat dissipation. A camera 16 is set up between the two light sources i 5 of the first source group and corresponds to the measurement area a of the biological reagent 8. Two fans 17 are set up in the accommodating space 1 1 for the heat dissipation holes 13 on both sides of the testing machine 1 to generate airflow, and the heat in the accommodating space 11 is divided into a plurality of heat dissipation holes 1 3 discharge. The data processing device 3 is connected to the camera 16 and performs image processing on the image of the biological reagent 8 obtained by the camera 16, and between the test line 8 2 and the control line 8 3 of the biological reagent 8 The light field intensity of the image in the light field measurement area B is calculated to obtain an average gray value as a reference threshold Graystandard, and the image of the test line 8 2 is vertically and horizontally divided into ^ regions C, and the base V threshold is Gray ^ ㈣ ^^ Compare with the control line inspection area d and test line 8 2 in each area C image light field intensity threshold τ η. If the average gray value of the image in any area c is Grayave is lower than the area threshold τn, the area c It is a positive reaction, and calculates the grayscale average value Grayp of all positive reaction areas, and calculates the level of the biochemical reaction concentration based on the average grayscale value. If the average grayscale value of all areas in the area is Grayave south of the area threshold ^, the biological reagent The test result was judged as a negative reaction. When taking the image of the biological reagent 8 with the camera 16, it is necessary to overcome the noise problem first. The present invention adopts a method of averaging multiple consecutive images to suppress the noise. °

Page 11569010 V. Description of the invention (7) L & v & i where S (X, Y) is the signal value of the image; the noise level of the i-th image acquisition of Ni (X, Y) is assumed to be the i-th image acquisition The overall noise is randomly distributed. The noise value of the overall image can be expressed as

The image miscellaneous image is as follows: L & v & I image is taken M times, and then its average value _ I Μ

Among them, Gray i (X, Y) is the gray value of each single point; M is the number of images.

a / MxSNR means that after the image has been acquired M times, the noise-to-noise ratio of the image will be increased by ^ times, and the noise is suppressed. Refer to Figure 6 for reference.

In addition, since the test line 8 2 on the biological reagent 8 is usually made by roll coating, it does not have very good uniformity, and the edges often have blurring and uneven concentration. To overcome this problem, the present invention performs the following operations on the image obtained by the camera 16 (as shown in FIG. 7): The operation that the binary image A is etched by B can be expressed as follows: ΑΘΒ = {χ ( Β) χ ^ Α}

Page 12569010 V. Description of the invention (8) The mathematical expression of the etching operation of gray image f and b can be expressed as follows: ίβφ, t) = min {f (x + s, i + y) -b ( x,: φ + χ), (^ + ¥); (χ, > 〇ε where Df and Db are the regions of f and b. If degradation becomes a function of a variable, the above formula becomes: (fBh) (s) = min {f (s + x)-ά (χ) | (^ + χ) ε Z? / and xe where B is a mask with a content of 1 in the range of 6 * 1. Further, when testing When the line 8 2 has an obvious reaction phenomenon, it can be clearly seen on the image of the light field measurement area B between the test line 8 2 and the control line 8 3 in the image of the test area A of the biological reagent 8 in FIG. 7. Figure 8 is the result of inverting the image of Figure 7 into a 3D three-dimensional figure, but when the test line 8 2 has no obvious reaction phenomenon, as shown in Figure 9 biological reagents It is difficult to determine the exact position of the test line 8 2 in the image of the tested area A. At this time, the data shown in Figures 10 and 11 can be obtained through the aforementioned calculations, and the test line can be clearly determined. 8 2 is indeed consistent position. The following tasks are performed in order: 1 · Check whether the control line 8 3 is normal to the control line 8 3 In the inspection area D, check whether the control line 8 3 is working normally. 2 · Calculate the geometric center position of the control line 8 3 and estimate the test line 8 2 and the location of the light field measurement area A.

In the geometry of 569010; test line test line 8 field summer measurement area line 8 2 detection field change, the light field measurement is offset from the camera 2 inspection area difference, if there is a threshold difference, each point = 1 ~ · 30 In the inspection, the value of the automatic adjustment of the valve in the average inspection zone of the environment B that needs to be inspected should be the existence of each of the points in Figure 12 during the occurrence of the photosensitive wafer phenomenon. Therefore, in order to reduce the cutting area, the light field amount is set to Pc; the light field measurement area A 2 and the center position of P 2 ° and the center distance difference is Δd. In the environment, the gray value of several changes in range The base threshold value is fixed, and the color drift is shown in the light cut area. The threshold value can be measured according to the light source. V. Explanation of the invention (9) Assuming that the geometric center position of the control line 8 3 is the control estimated by P i Line 8 3 and Μ rules; 3. Light field measurement In order to guarantee the error intensity measurement produced by each detection of low interference, the light value of the detection hypothesis is used as the test 4 · According to the theoretical error of light intensity, the overall color and Test line 8 The light field intensity of the fixed difference is a fixed area. The average gray value of the test line inspection area is different, and the primes are kept fixed. The light field at a specific position is lowered as the lightness. However, because of the lighting phenomenon of the light source system, the hidden light field intensity C of the extracted light field measurement has the light field intensity of the near area c 1 and the area c ^ light field changes' automatic adjustment error. It is assumed that ; Light intensity difference of city in flat divided area of light measurement area

Page 14569010 V. Description of the invention (? Ο) 2-'· standard +, f " J = 1 ··· 30 5 · Test line for detecting biological reagents In this example, the test line 8 2 is tested Divided into 30 small areas 'Each small area sets a different threshold depending on the light field' to test whether the test line 8 2 reacts. In addition, the number of pixels with positive reactions in the segmented area C in the inspection area often is smaller than the number of pixels with negative reactions. In order to avoid the average gray value of the segmented area C being misjudged as a negative reaction, When the gray value is averaged, the gray value of the pixel that may be a positive response will be increased to improve the accuracy of the judgment. Assume the area threshold rule, Η = 1 ~ 30; the area average gray value; the gray value of the pixel less than the area threshold; the number of pixels less than the area threshold; Gray value of pixels greater than regional threshold ^ Butterfly (X, magic; number of pixels greater than regional threshold

Gmyt

Num ^ +

GraykJ ^ y, S < Grny

6 · Judging the reaction result When the reaction color Gr of any of the divided areas in the test area is lower than the area threshold, and π = 1 ~ 30, it is a positive reaction, and all positives are calculated.

Page 15 569010 V. Description of the invention ο) The average gray value P of the color in the reaction area, and the level of the biochemical reaction concentration is calculated based on the average gray value. If there is no response, it is judged as a negative reaction. Positive reaction: Assume that there is a segmented area showing a positive response; the color of the segmented area showing a positive response; the area threshold 7 ;, η = 1 ~ 30; the color of all positive reaction areas is gray. The average gray value of the measurement area of the average light field is ^; the grade of the biochemical reaction concentration is Z. d; the grade interval is η

Level

Num, where to apply ~> 0 and <7; Negative reaction: The average gray value of the negative area is higher than the inter-area value, then the detection result of the HCV reagent according to the present invention has 2 1 ==, should (Posltive) : There is a kind of w " < Jiu Zijian = ^ f indicates that the control line appears with a color band, and at the same time the test line also appears with a color band at the same time, indicating that the test subject's blood, and the main device are wide. The color of the ribbon, the dish, the bite is also the owner = blood, the f of the month has Antl-HCV and the anti-steam is low. / Clothing times can represent the antibody titer (t i ter) in serum

Page 16569010 —- ~~~~~~ One __________ 5. Explanation of the invention (12) 2 · Negative: When the test result shows that only the control line shows a color band, it means that the test subject's serum does not contain Anti-HCV antibodies. 3 · Invalid: If there is no color band in the test result or no color band appears on the control line ’this means that the reagent has expired, expired, or operated improperly, and another test is required. The data processing device 3 of the present invention can be a personal computer, and the image is received to the computer through a USB2.0 transmission interface (the transmission rate is up to 4800 Mbps). Using the automatic visual inspection rule proposed by the present invention, the biochemical reaction can be identified in real time to determine whether Positive or negative. If it is a positive response, then the level of the reaction concentration is calculated to provide objective and accurate data ′ as an important basis for medical personnel to diagnose hepatitis C. In terms of operation, the user only needs to place the biological reagent 8 to be tested in the slide 1 of the testing machine 1 and then press the button in the software of the data processing device 3 to start the detection. The system will automatically recognize the biochemical reaction , And output the result to the table, and mark the area of the positive reaction in the image. As shown in Figure 13 is the result of detecting a negative reaction; as shown in Figures 14 to 16 is the result of detecting a positive reaction. Among them. Brother 1 4 SI system shows no biochemical reaction 丨 Chen degree grading 丨 = 50 state. Figure 15 shows the state of _; = 75 in the biochemical reaction concentration level. Fig. 16 shows a state in which the concentration level of the biochemical reaction is ㈣ = 1 00. In summary, the present invention uses a camera 16 to capture the biological agent 8

569010 V. Description of the invention (13) Measure the image of area A, and then cut the image of part 2 of the test line 8 of biological reagent 8 into n areas. By comparing the thresholds of each area, the biological test can be accurately judged. The effectiveness of the test results. The disclosure of the embodiments described above is used to illustrate the present invention, and is not intended to limit the present invention. Therefore, any change in the value or replacement of equivalent components should still belong to the scope of the present invention. From the above detailed description, those skilled in the art can understand that the present invention can indeed achieve the aforementioned purpose, and it has indeed complied with the provisions of the Patent Law, and filed a patent application.

Page 18569010 Brief description of the drawings [Simplified description of the drawings] Figure 1 is a schematic diagram of the appearance of a biological reagent. Figure 2 is a schematic diagram of the configuration of the detection system of the present invention. Figure 3 is one of the schematic diagrams of the structure of the testing machine of the present invention. Figure 4 is the second schematic diagram of the structure of the testing machine of the present invention. Figure 5 is the schematic diagram of the biological reagent image detection area of the present invention. Figure 6 is the single-point schematic diagram of the present invention when filtering out noise. Image 8 of the biological reagent 1 obtained by the camera of the present invention. Figure 8 is the three-dimensional coordinate value of the image of the biological reagent 1 obtained by the camera of the present invention. Figure 9 is the image of the biological reagent 2 obtained by the camera of the present invention. The three-dimensional coordinate values of the image. Figure 1 is a binary value figure obtained after the binary image of the biological reagent in the present invention is obtained through a binary operation. Figure 12 is a schematic diagram of the light field distribution in the present invention. Display of the day when the negative reaction result is detected in the invention. Figure 14 shows the concentration level of the biochemical reaction when the positive reaction is detected by the present invention, iewi = 50. The display screen in the state of Figure 15 shows the result of the biochemical detection of the positive reaction by the present invention. = Density level should display surface of the first day 16 the state of FIG. 75 positive biochemical reaction system ώνβ = concentration level state of the display screen 100 [FIG No. Description The present invention is detected

Page 19 569010 Schematic description

Slide rail 1 4 Light source 1 5 Camera 1 6 Data processing device 3 Operating area 8 1 Control line 8 3 Light field measurement area B Control line inspection area D Positioning auxiliary rod 1 4 1 Radiating fin 1 5 1 Fan 1 7 Biological reagent 8 Test line 8 2 Test area A Area C, C 1, C 2

Page 20

Claims (1)

569010 VI. Scope of patent application1. An automatic biological reagent detection system, which includes: a testing machine, the testing machine has an accommodation space, and the testing machine has a door that can be opened and closed for biological reagents Enter and exit the accommodating space, and the testing machine has several cooling holes for air circulation; a sliding handle, which is erected in the accommodating space of the testing machine, and a positioning aid is provided at the end of the slide rail. A rod for the biological reagent to enter and exit the accommodation space through the slider, and the positioning auxiliary rod provides the positioning function; a light source group, which is composed of two light sources, which are spaced apart from the corresponding biological above the slide rail At the test area of the reagent, the two light sources are arranged symmetrically and inclined, and the direction in which the two light sources are inclined is perpendicular to the extending direction of the slider. In addition, heat radiation fins 贴 are attached to each light source to strengthen it. Radiating effect; a camera is set between two light sources of the light source group and corresponds to a test area of biological reagents; more than one fan is set in the In the installation space, the heat in the accommodation space is exhausted through several heat dissipation holes to generate airflow. A data processing device is connected to the camera and image-processes the biological reagent image obtained by the camera. Use a mask with a range of 1 in the range of 6 * 1 to perform the #engraving operation to eliminate the noise of the dyeing effect on the reagent, and perform the following tasks in order; (1) · Check whether the control line is normal in the control line inspection area Check whether the control line works normally; Page 21 569010 VI. Patent application scope (2) · Calculate the geometric center position of the control line, and estimate the position of the test line and the light field measurement area A. Measure the light field between the test line of the biological reagent and the control line. Area image light% intensity calculation to obtain the average gray value as the threshold value Graystandard 'and divide the test line image horizontally and vertically into regions' and use the threshold value Graystandard and the control line part and each area of the test line image light The% intensity threshold τ η is compared. When the average gray value Grayave of any area image is lower than the area threshold τn, the area is a positive reaction, and the average gray level grayp of all positive reaction areas is calculated. The average value of the gray level is used to calculate the level of the biochemical reaction concentration. The average gray value of Grayave in all the areas on the right is higher than the threshold value of the area ^^ The detection result of the biological reagent is judged as a negative reaction; if the judgment and processing rules are not: (1) The threshold value of the light field intensity of each area is equal to the fixed difference between the light field intensity of the light field measurement area and ω η. Then, the light field intensity threshold value of each area τη = the reference valve. If the value is Graystandard + ω ^, the data processing device will calculate the threshold value τη of each area accordingly, and then compare the calculated threshold value τη of each area with the image light field intensity of the test line to reduce the The error produced; (2) The average gray value of each region desired by the data processing device, Grayave, is less than the regional threshold value of the gray value of the pixel, Grayi0w (X, 乂): the number of pixels smaller than the regional threshold value, NumlQW; Grayscale weight value ω of the pixel smaller than the regional threshold; Grayhigh (X, y) of the gray value of the pixel larger than the regional threshold; the number of pixels larger than the regional threshold 569010 VI. The scope of patent application NU m high? The regional average degree value, where Omy ^ ix.y) ^ < 〇Γ〇γ ^ 9ν), thereby increasing the grayscale weight value of the pixel that may be a positive reaction, to Improve the accuracy of judgment; (3) The data processing device calculates the concentration level of biochemical reaction Level Hiichi, 'Σ Gray-The formula is ^ = Bao Zhou, where% > 0, Gray ^ <7;, and Nump is the number of blocks showing a positive reaction, and η is the interval of concentration levels of biochemical reactions. 2. The biological reagent automatic detection system according to item 1 of the scope of the patent application, wherein each light source of the light source group is a yellow light backlight board. 3. The biological reagent automatic detection system according to item 1 of the scope of the patent application, wherein both sides of the detection machine have several heat dissipation holes, and the accommodation space is close to the heat dissipation holes on both sides of the test machine, respectively. Equipped with a fan. Page 23