Tongue swab sample fluorescence detection method for mycobacterium tuberculosis based on image processing

Abstract

The invention discloses an image processing-based tongue swab sample fluorescence detection method for Mycobacterium tuberculosis, and relates to the technical field of image processing, the image processing-based tongue swab sample fluorescence detection method comprises a flora monitoring module, an image analysis module and a concentration determination module, the flora monitoring module is used for identifying a flora culture solution and monitoring flora on the flora culture solution in real time, and the concentration determination module is used for determining the concentration of Mycobacterium tuberculosis; the image analysis module is used for carrying out straight division processing and flora contour fitting on a shot image, and the concentration determination module is used for analyzing the flora size, the shape and the life cycle of mycobacteria in image data to carry out targeted fluorescent agent concentration adjustment on florae with different growth conditions. The flora monitoring module comprises an image monitoring unit and a flora culture solution recognition module, the image monitoring unit is electrically connected with the flora culture solution recognition module and the flora recognition module, and the flora culture solution recognition module is electrically connected with the flora recognition module.

Description

Technical Field

[0001] The invention relates to the technical field of image processing, and in particular to a method for fluorescence detection of Mycobacterium tuberculosis by tongue swab samples based on image processing. Background Art

[0002] Fluorescence detection of Mycobacterium tuberculosis in tongue swab samples is a laboratory method for diagnosing tuberculosis. Fluorescence detection technology can quickly and sensitively identify the presence of Mycobacterium tuberculosis, thereby achieving early diagnosis of tuberculosis. During the growth and division of Mycobacterium tuberculosis, the absorption of fluorescent agents will also change. In the early stages of growth and division, the individuals are small and the rod-shaped divergence characteristics are not obvious. With the rapid proliferation of bacteria, the number of bacteria increases and they will present a typical rod-shaped divergence structure. As the bacteria die, the division rate will slow down.

[0003] When the existing mycobacterium flora is taken out of the culture room for observation and growth experiment, the same concentration of fluorescent agent is uniformly added for coloring. However, the life cycle of the flora is different. Studies have shown that the concentration of fluorescent agent adapted to different flora life cycles is also different. As the bacterial life cycle continues, its absorption capacity of fluorescent agent decreases, so more fluorescent agent is needed for better observation. Mycobacterium flora in the early stage of the life cycle will absorb excessive fluorescent agent, resulting in unclear observation of details. Therefore, it is necessary to design a real-time control method for fluorescence detection of mycobacterium tuberculosis in tongue swab samples based on image processing. Summary of the invention

[0004] The object of the present invention is to provide a method for fluorescence detection of Mycobacterium tuberculosis from tongue swab samples based on image processing, so as to solve the problems raised in the above background technology.

[0005] In order to solve the above technical problems, the present invention provides the following technical solutions: a method for fluorescent detection of Mycobacterium tuberculosis in tongue swab samples based on image processing, comprising a flora monitoring module, an image analysis module, and a concentration determination module. The flora monitoring module is used to identify the flora culture fluid and to perform real-time monitoring of the flora on the flora culture fluid. The image analysis module is used to perform straight division processing on the captured image and fit the flora contour. The concentration determination module is used to analyze the flora size, Mycobacterium shape and life cycle in the image data to make targeted adjustments to the fluorescent agent concentration for flora with different growth conditions.

[0006] According to the above technical solution, the flora monitoring module includes an image monitoring unit, a flora culture solution recognition module, and a flora recognition module. The image monitoring unit is electrically connected to the flora culture solution recognition module and the flora recognition module. The flora culture solution recognition module is electrically connected to the flora recognition module. The image monitoring unit is used to obtain bacterial culture liquid image data, the bacterial culture liquid identification module is used to identify the bacterial culture liquid using image recognition technology to determine the bacterial culture liquid in which the bacterial colony grows, and the bacterial colony identification module is used to identify the bacterial colony on the bacterial culture liquid and determine the position of the bacterial colony in the bacterial culture liquid; The image analysis module includes a coordinate establishment module, a growth area division module, a position marking module, a flora outline drawing module, and an interval photographing module. The interval photographing module is electrically connected to the position marking module, and the position marking module is electrically connected to the flora outline drawing module. The coordinate establishment module is electrically connected to the growth area division module and the position marking module in sequence. The coordinate establishment module is used to generate a culture area map for the monitored flora culture liquid image. The growth area division module is used to intersect the culture area map with straight lines and vertical lines at the same interval size, and set boundaries at the intersection points. The position marking module is used to record the point coordinates of the flora image elements. The flora outline drawing module is used to identify the outline of the flora image and draw a simplified flora outline. The interval photographing module is used to take pictures and record the image at a certain period. The concentration determination module includes a time marking module, a bacterial colony size analysis module, a growth speed calculation module, an individual shape judgment module, a concentration calculation module, and a weight determination module. The time marking module is electrically connected to the growth speed calculation module, the growth speed calculation module is electrically connected to the concentration calculation module, and the bacterial colony size analysis module and the individual shape judgment module are electrically connected to the bacterial colony outline drawing module. The time stamp module is used to calculate the time as the basis for other data, the bacterial colony size analysis module is used to analyze the point coordinates of the bacterial colony image elements and the outline size of the bacterial colony, the growth rate calculation module is used to calculate the life cycle of the bacterial colony based on the changes in shape and size during the image recording cycle, the individual shape judgment module is used to judge the shape of individual mycobacteria in the bacterial colony in combination with the image elements, the concentration calculation module is used to reduce the concentration of fast-growing bacterial colonies, and the weight determination module is used to adjust the weight of the influence of the size of the bacterial colony and the shape of the individual on the life cycle of the bacterial colony according to the use time of the culture solution.

[0007] According to the above technical solution, the working method of the system is: S1, taking mycobacteria and dividing them into several portions of similar size, using an image monitoring unit to photograph mycobacterial colonies of the same initial size, and generating coordinates of each intersection as the growth position of the colony, and counting the use time of the culture solution; S2, taking photos and recording the image information of the intersection at a certain period, and not recording the image information at other times, and drawing the outline of the bacterial colony according to whether the image elements of the bacterial colony appear at the coordinate point; S3, calculating the size of the bacterial colony's outline in combination with the bacterial colony's outline scanning image to obtain the bacterial colony's division speed, and analyzing the bacterial colony's shape to obtain the bacterial colony's growth stage; S4. According to the use time of the culture medium, the weight of the shape of the mycobacterium on the life cycle of the bacterial community is adjusted, and the life cycle of the bacterial community is calculated; S5. Adjust the concentration of the fluorescent agent according to the life cycle of the bacterial colony so that the observation conditions of each bacterial colony are adjusted to a similar state, which is convenient for unified observation and comparison.

[0008] According to the above technical solution, in S3, the method for calculating the outline size of the bacterial colony is: S3-1: After the flora recognition module identifies the flora outline, the flora outline drawing module marks and locks the pixel points on the shape edge of the flora image, and smoothly connects the adjacent pixel points. The image grid is at the base distance between the flora and the image monitoring unit. The base unit area is ; S3-2: Calculate the number of image grids that are surrounded by smooth lines , which is the image size of the bacterial colony, and the distance value perceived by the image distance , quantify the bacterial colony growth rate at the current distance , at the same time The bigger the size, the faster the splitting speed. is the conversion factor between the image size of the bacterial colony and its reproduction rate.

[0009] According to the above technical solution, in S3, the specific method for analyzing the shape of the flora is: S3-3. From the microscope image, the horizontal extreme point coordinates of the Mycobacterium individuals in the bacterial colony are: , and connect the marked points to draw a horizontal line for the middle mycobacterium; S3-4, from the microscope image, the vertical extreme point coordinates of the mycobacterium individuals in the bacterial colony are , and connect the marked points to draw a longitudinal line of mycobacteria; S3-5. Perform shape analysis on all mycobacterium individuals in the bacterial community and calculate the average rod-shaped divergence characteristics of mycobacterium individuals in the bacterial community, i.e., the growth stage value , , , , , , , , , are the coordinate values ​​of the marked points. The larger the growth stage, the closer it is to maturity.

[0010] According to the above technical solution, in S4, the life cycle of the bacterial community The calculation method is as follows: Since the proportion of bacterial flora at different stages of development and the size of mature shapes are different, the bacterial flora reproduction speed and the value of bacterial flora growth stage are different. Neither can fully and accurately reflect the life cycle of the bacterial community , The bacterial colony growth rate and the growth stage values ​​of the flora The comprehensive value of , is the weight coefficient of bacterial colony growth rate, is the weight coefficient of the bacterial colony growth stage, which is determined based on actual experience. In the early stage of using bacteria in the culture medium, Maximum, with the continuous use of culture medium, It will decrease in direct proportion to the usage time.

[0011] According to the above technical solution, in S5, the specific method for adjusting the concentration of the fluorescent agent is: set up an injection point for each coordinate point, and take pictures at each time point according to the life cycle of the bacterial colony in the normal growth stage of the bacterial colony. Perform preliminary setting of fluorescent agent concentration, i.e. standard life cycle of bacterial colonies at different time points They are , corresponding to the standard concentration of fluorescent agent , when the life cycle of a bacterial community Below this time point When the concentration of fluorescent agent is reduced , otherwise it increases .

[0012] According to the above technical solution, in S5, the specific calculation method of the concentration of the fluorescent agent is: ,in is the concentration conversion factor of the fluorescent agent.

[0013] Compared with the prior art, the beneficial effects achieved by the present invention are as follows: the present invention uses image analysis to conduct comprehensive statistics on the division rate and growth stage of the Mycobacterium flora to obtain the life cycle of the flora. This method of calculating the life cycle takes two factors into consideration and will not cause excessive deviations due to a single judgment. At the same time, the concentration of the fluorescent agent can be adjusted to make it as consistent as possible with the life cycle, which is convenient for accurate observation. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings: Figure 1 It is a schematic diagram of the overall module structure of the present invention. DETAILED DESCRIPTION

[0015] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0016] See also Figure 1 The present invention provides a technical solution: a method for fluorescent detection of tuberculosis mycobacterium from a tongue swab sample based on image processing, comprising a flora monitoring module, an image analysis module, and a concentration determination module, wherein the flora monitoring module is used to identify the flora culture solution and to monitor the flora on the flora culture solution in real time, the image analysis module is used to perform straight division processing on the captured image and fit the flora contour, and the concentration determination module is used to analyze the flora size, mycobacterium shape and life cycle in the image data to perform targeted fluorescent agent concentration adjustment for flora with different growth conditions; The flora monitoring module includes an image monitoring unit, a flora culture solution recognition module, and a flora recognition module. The image monitoring unit is electrically connected to the flora culture solution recognition module and the flora recognition module. The flora culture solution recognition module is electrically connected to the flora recognition module. The image monitoring unit is used to obtain the image data of the bacterial culture solution, the bacterial culture solution recognition module is used to identify the bacterial culture solution using the image recognition technology, and determine the bacterial culture solution in which the bacterial community grows, and the bacterial community recognition module is used to identify the bacterial community on the bacterial culture solution and determine the position of the bacterial community in the bacterial culture solution; The image analysis module includes a coordinate establishment module, a growth area division module, a position marking module, a bacterial colony outline drawing module, and an interval photographing module. The interval photographing module is electrically connected to the position marking module, and the position marking module is electrically connected to the bacterial colony outline drawing module. The coordinate establishment module is electrically connected to the growth area division module and the position marking module in sequence. The coordinate establishment module is used to generate a culture area map for the monitored bacterial colony culture liquid image. The growth area division module is used to intersect the culture area map with straight lines and vertical lines at the same interval size, and set boundaries at the intersection points. The position marking module is used to record the point coordinates of the bacterial colony image elements. The bacterial colony outline drawing module is used to identify the outline of the bacterial colony image and draw a simplified bacterial colony outline. The interval photographing module is used to take pictures and record the image at a certain period. The concentration determination module includes a time marking module, a bacterial colony size analysis module, a growth speed calculation module, an individual shape judgment module, a concentration calculation module, and a weight determination module. The time marking module is electrically connected to the growth speed calculation module, the growth speed calculation module is electrically connected to the concentration calculation module, and the bacterial colony size analysis module and the individual shape judgment module are electrically connected to the bacterial colony outline drawing module. The time stamp module is used to calculate the time as the basis for other data. The bacterial colony size analysis module is used to analyze the bacterial colony outline size according to the point coordinates of the bacterial colony image elements. The growth rate calculation module is used to calculate the bacterial colony life cycle according to the changes in shape and size during the image recording cycle. The individual shape judgment module is used to judge the shape of individual mycobacteria in the bacterial colony in combination with the image elements. The concentration calculation module is used to reduce the concentration of fast-growing bacterial colonies. The weight determination module is used to adjust the weight of the influence of the size of the bacterial colony and the shape of the individual on the bacterial colony life cycle according to the use time of the culture solution. The system works as follows: S1, taking mycobacteria and dividing them into several portions of similar size, using an image monitoring unit to photograph mycobacterial colonies of the same initial size, and generating coordinates of each intersection as the growth position of the colony, and counting the use time of the culture solution; S2, taking photos and recording the image information of the intersection at a certain period, and not recording the image information at other times, and drawing the outline of the bacterial colony according to whether the image elements of the bacterial colony appear at the coordinate point; S3, calculating the size of the bacterial colony's outline in combination with the bacterial colony's outline scanning image to obtain the bacterial colony's division speed, and analyzing the bacterial colony's shape to obtain the bacterial colony's growth stage; S4. According to the use time of the culture medium, the weight of the shape of the mycobacterium on the life cycle of the bacterial community is adjusted, and the life cycle of the bacterial community is calculated; S5. Adjust the concentration of the fluorescent agent according to the life cycle of the bacterial colony so that the observation conditions of each bacterial colony are adjusted to a similar state, which is convenient for unified observation and comparison; In S3, the method for calculating the outline size of the bacterial colony is: S3-1: After the flora recognition module identifies the flora outline, the flora outline drawing module marks and locks the pixel points on the shape edge of the flora image, and smoothly connects the adjacent pixel points. The image grid is at the base distance between the flora and the image monitoring unit. The base unit area is ; S3-2: Calculate the number of image grids that are surrounded by smooth lines , which is the image size of the bacterial colony, and the distance value perceived by the image distance , quantify the bacterial colony growth rate at the current distance , at the same time The bigger the size, the faster the splitting speed. is the conversion factor between the image size of the bacterial colony and its reproduction speed; In S3, the specific method for analyzing the shape of the bacterial colony is: S3-3. From the microscope image, the horizontal extreme point coordinates of the Mycobacterium individuals in the bacterial colony are: , and connect the marked points to draw a horizontal line for the middle mycobacterium; S3-4, from the microscope image, the vertical extreme point coordinates of the mycobacterium individuals in the bacterial colony are , and connect the marked points to draw a longitudinal line of mycobacteria; S3-5. Perform shape analysis on all mycobacterium individuals in the bacterial community and calculate the average rod-shaped divergence characteristics of mycobacterium individuals in the bacterial community, i.e., the growth stage value , , , , , , , , , are the coordinate values ​​of the marked points. The larger the growth stage, the closer to maturity; In S4, the life cycle of the bacterial community The calculation method is: Since the proportion of bacterial colonies at different stages of development and their shapes and sizes after maturity are different, the bacterial colony reproduction speed and the growth stage values ​​of the flora Neither can fully and accurately reflect the life cycle of the bacterial community , The bacterial colony growth rate and the growth stage values ​​of the flora The comprehensive value of , is the weight coefficient of bacterial colony growth rate, is the weight coefficient of the bacterial colony growth stage, which is determined based on actual experience. In the early stage of using bacteria in the culture medium, Maximum, with the continuous use of culture medium, It will decrease in direct proportion to the time of use; In S5, the specific method for adjusting the concentration of the fluorescent agent is: set up an injection point for each coordinate point, and take pictures at each time point according to the life cycle of the bacterial colony in the normal growth stage of the bacterial colony. Perform preliminary setting of fluorescent agent concentration, i.e. standard life cycle of bacterial colonies at different time points They are , corresponding to the standard concentration of fluorescent agent , when the life cycle of a bacterial community Below this time point When the concentration of fluorescent agent is reduced , otherwise it increases ; In S5, the specific calculation method of the concentration of the fluorescent agent is: ,in is the concentration conversion factor of the fluorescent agent.

[0017] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0018] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for fluorescence detection of Mycobacterium tuberculosis from tongue swab samples based on image processing, characterized in that: The method adopts a detection system to work, which includes a flora monitoring module, an image analysis module, and a concentration determination module. The flora monitoring module is used to identify the flora culture solution and monitor the flora on the flora culture solution in real time. The image analysis module is used to flatten the captured image and fit the flora contour. The concentration determination module is used to analyze the flora size, mycobacterium shape and life cycle in the image data to make targeted adjustments to the fluorescent agent concentration for flora with different growth conditions.

2. The method for fluorescence detection of Mycobacterium tuberculosis from tongue swab samples based on image processing according to claim 1, characterized in that: The flora monitoring module includes an image monitoring unit, a flora culture solution recognition module, and a flora recognition module. The image monitoring unit is electrically connected to the flora culture solution recognition module and the flora recognition module. The flora culture solution recognition module is electrically connected to the flora recognition module. The image monitoring unit is used to obtain bacterial culture liquid image data, the bacterial culture liquid identification module is used to identify the bacterial culture liquid using image recognition technology to determine the bacterial culture liquid in which the bacterial colony grows, and the bacterial colony identification module is used to identify the bacterial colony on the bacterial culture liquid and determine the position of the bacterial colony in the bacterial culture liquid; The image analysis module includes a coordinate establishment module, a growth area division module, a position marking module, a flora outline drawing module, and an interval photographing module. The interval photographing module is electrically connected to the position marking module, and the position marking module is electrically connected to the flora outline drawing module. The coordinate establishment module is electrically connected to the growth area division module and the position marking module in sequence. The coordinate establishment module is used to generate a culture area map for the monitored flora culture liquid image. The growth area division module is used to intersect the culture area map with straight lines and vertical lines at the same interval size, and set boundaries at the intersection points. The position marking module is used to record the point coordinates of the flora image elements. The flora outline drawing module is used to identify the outline of the flora image and draw a simplified flora outline. The interval photographing module is used to take pictures and record the image at a certain period. The concentration determination module includes a time marking module, a bacterial colony size analysis module, a growth speed calculation module, an individual shape judgment module, a concentration calculation module, and a weight determination module. The time marking module is electrically connected to the growth speed calculation module, the growth speed calculation module is electrically connected to the concentration calculation module, and the bacterial colony size analysis module and the individual shape judgment module are electrically connected to the bacterial colony outline drawing module. The time stamp module is used to calculate the time as the basis for other data, the bacterial colony size analysis module is used to analyze the point coordinates of the bacterial colony image elements and the outline size of the bacterial colony, the growth rate calculation module is used to calculate the life cycle of the bacterial colony based on the changes in shape and size during the image recording cycle, the individual shape judgment module is used to judge the shape of individual mycobacteria in the bacterial colony in combination with the image elements, the concentration calculation module is used to reduce the concentration of fast-growing bacterial colonies, and the weight determination module is used to adjust the weight of the influence of the size of the bacterial colony and the shape of the individual on the life cycle of the bacterial colony according to the use time of the culture solution.

3. The method for fluorescence detection of Mycobacterium tuberculosis from tongue swab samples based on image processing according to claim 2, characterized in that: The system works as follows: S1, taking mycobacteria and dividing them into several portions of similar size, using an image monitoring unit to photograph mycobacterial colonies of the same initial size, and generating coordinates of each intersection as the growth position of the colony, and counting the use time of the culture solution; S2, taking photos and recording the image information of the intersection at a certain period, and not recording the image information at other times, and drawing the outline of the bacterial colony according to whether the image elements of the bacterial colony appear at the coordinate point; S3, calculating the size of the bacterial colony's outline in combination with the bacterial colony's outline scanning image to obtain the bacterial colony's division speed, and analyzing the bacterial colony's shape to obtain the bacterial colony's growth stage; S4. According to the use time of the culture medium, the weight of the shape of the mycobacterium on the life cycle of the bacterial community is adjusted, and the life cycle of the bacterial community is calculated; S5. Adjust the concentration of the fluorescent agent according to the life cycle of the bacterial colony so that the observation conditions of each bacterial colony are adjusted to a similar state, which is convenient for unified observation and comparison.

4. The method for fluorescence detection of Mycobacterium tuberculosis from tongue swab samples based on image processing according to claim 3, characterized in that: In S3, the method for calculating the outline size of the bacterial colony is: S3-1: After the flora recognition module identifies the flora outline, the flora outline drawing module marks and locks the pixel points on the shape edge of the flora image, and smoothly connects the adjacent pixel points. The image grid is at the base distance between the flora and the image monitoring unit. The base unit area is ; S3-2: Calculate the number of image grids that are surrounded by smooth lines , which is the image size of the bacterial colony, the distance value perceived by the image distance, and the quantification of the bacterial colony reproduction speed at the current distance , at the same time The bigger the size, the faster the splitting speed. is the conversion factor between the image size of the bacterial colony and its reproduction rate.

5. The method for fluorescence detection of Mycobacterium tuberculosis from tongue swab samples based on image processing according to claim 4, characterized in that: In S3, the specific method for analyzing the shape of the bacterial colony is: S3-3. From the microscope image, the horizontal extreme point coordinates of the Mycobacterium individuals in the bacterial colony are: , and connect the marked points to draw a horizontal line for the middle mycobacterium; S3-4, from the microscope image, the vertical extreme point coordinates of the mycobacterium individuals in the bacterial colony are , and connect the marked points to draw a longitudinal line of mycobacteria; S3-5. Perform shape analysis on all mycobacterium individuals in the bacterial community and calculate the average rod-shaped divergence characteristics of mycobacterium individuals in the bacterial community, i.e., the growth stage value , ， , , , , , , , are the coordinate values ​​of the marked points. The larger the growth stage, the closer it is to maturity.

6. The method for fluorescence detection of Mycobacterium tuberculosis from tongue swab samples based on image processing according to claim 5, characterized in that: In S4, the life cycle of the bacterial community The calculation method is: Since the proportion of bacterial colonies at different stages of development and their shapes and sizes after maturity are different, the bacterial colony reproduction speed and the growth stage values ​​of the flora Neither can fully and accurately reflect the life cycle of the bacterial community , The bacterial colony growth rate and the growth stage values ​​of the flora The comprehensive value of , is the weight coefficient of bacterial colony growth rate, is the weight coefficient of the bacterial colony growth stage, which is determined based on actual experience. In the early stage of using bacteria in the culture medium, Maximum, with the continuous use of culture medium, It will decrease in direct proportion to the usage time.

7. The method for fluorescence detection of Mycobacterium tuberculosis from tongue swab samples based on image processing according to claim 6, characterized in that: In S5, the specific method for adjusting the concentration of the fluorescent agent is: setting an injection point for each coordinate point, taking pictures at each time point according to the life cycle of the bacterial colony in the normal growth stage of the bacterial colony. Perform preliminary setting of fluorescent agent concentration, i.e. standard life cycle of bacterial colonies at different time points They are , corresponding to the standard concentration of fluorescent agent , when the life cycle of a bacterial community Below this time point When the concentration of fluorescent agent is reduced , otherwise it increases .

8. The method for fluorescence detection of Mycobacterium tuberculosis from tongue swab samples based on image processing according to claim 7, characterized in that: In S5, the specific calculation method of the concentration of the fluorescent agent is: ,in is the concentration conversion factor of the fluorescent agent.

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