Assessment method and apparatus for immunoblot detection membrane strip, and storage medium and electronic device

Abstract

Provided in the present invention are an assessment method and apparatus for an immunoblot detection membrane strip, and a storage medium and an electronic device, wherein the immunoblot detection membrane strip is provided with a plurality of reaction lines. The assessment method comprises the following steps: acquiring a standard color image of an immunoblot detection membrane strip; converting the standard color image of the immunoblot detection membrane strip into a standard grayscale image; acquiring a grayscale value of each reaction line in the standard grayscale image; according to the grayscale value of each reaction line in the standard grayscale image and a preset conversion relationship between grayscale values and reflectivities, acquiring the reflectivity of each reaction line; and according to the reflectivity of each reaction line, quantitatively assessing characteristics of a protein, which is detected by the immunoblot detection membrane strip. The assessment method has the advantages of being empirical, specific, objective and traceable.

Description

Immunoblotting detection membrane strip evaluation method and device, storage medium and electronic equipment Technical Field

[0001] The present invention belongs to the technical field of immunoblotting detection, and in particular relates to an evaluation method and device for an immunoblotting detection membrane strip, a storage medium and an electronic device. Background Art

[0002] Immunoblotting, also known as protein blotting, is a method for detecting a certain protein in a complex sample based on the specific binding of antigens and antibodies. Due to its advantages such as large analytical capacity, high sensitivity, and strong specificity, it has become one of the most commonly used methods for detecting protein characteristics, expression, and distribution.

[0003] Both manual and automated immunoblotting are widely used in the medical field. However, manual testing can only provide qualitative analysis, and the results are subject to subjectivity, low efficiency, and error-proneness. Existing automated testing methods mostly produce qualitative or semi-quantitative results, which are characterized by low accuracy. In practice, this makes it difficult for doctors to formulate more reasonable, effective, and accurate treatment plans based on these qualitative or semi-quantitative results.

[0004] Although there are some methods that use grayscale values ​​to quantify the results of immunoblotting analysis, grayscale values ​​only exist in the image processing process and cannot be traced, which does not meet the requirement of the medical field that medical data can be traced.

[0005] Summary of the Invention

[0006] The purpose of the present invention is to provide an evaluation method and device for immunoblotting detection membrane strips, a storage medium and an electronic device, aiming to achieve quantitative detection of immunoblotting detection membrane strips and make the detection results traceable.

[0007] To achieve the above object, the present invention provides a quantitative evaluation method for an immunoblot detection membrane strip, wherein the immunoblot detection membrane strip has multiple reaction lines, and the evaluation method comprises the following steps:

[0008] Obtaining a standard color image of the immunoblotting detection membrane strip;

[0009] Converting the standard color image of the immunoblotting detection membrane strip into a standard grayscale image;

[0010] Obtaining a grayscale value of each reaction line in the standard grayscale image;

[0011] Obtaining the reflectivity of each of the reaction lines according to the grayscale value of each of the reaction lines in the standard grayscale image and a preset conversion relationship between the grayscale value and the reflectivity;

[0012] The characteristics of the protein detected by the immunoblotting detection membrane strip are quantitatively evaluated according to the reflectance of each reaction line.

[0013] Optionally, a conversion relationship between grayscale value and reflectivity is obtained by setting grayscale values ​​and corresponding reflectivity of multiple baselines on the quality control test paper;

[0014] And, the standard color image is obtained by photographing the immunoblot detection membrane strip under a standard shooting environment; the standard shooting environment refers to a shooting environment in which the quality control test paper is photographed to obtain a quality control test paper color image, and the grayscale value of each baseline in the quality control test paper grayscale image corresponding to the quality control test paper color image is equal to the corresponding set grayscale value.

[0015] Optionally, the step of obtaining the standard shooting environment includes:

[0016] Taking a picture of the quality control test paper to obtain a color image of the quality control test paper;

[0017] Converting the color image of the quality control test paper into a grayscale image of the quality control test paper;

[0018] Obtaining the grayscale value of each baseline on the grayscale image of the quality control test paper;

[0019] Determine whether the grayscale value of each baseline on the grayscale image of the quality control test paper is equal to the corresponding set grayscale value. If so, use the current shooting environment as the standard shooting environment; if not, adjust at least one of the intensity of the light emitted by the light source used for lighting, the incident angle of the light, and the exposure time of the camera.

[0020] Optionally, the step of obtaining the grayscale value of each baseline on the grayscale image of the quality control test paper includes:

[0021] Extracting a first strip image including all the reference lines from the grayscale image of the quality control test paper;

[0022] Counting the total number of pixels of each baseline;

[0023] Accumulating the grayscale values ​​of all pixels of each of the reference lines to obtain a total grayscale value of each of the reference lines;

[0024] The ratio of the total grayscale value of the corresponding reference line to the total number of pixels is calculated as the grayscale value of the corresponding reference line.

[0025] Optionally, the step of cutting out a first strip image including all the reference lines from the grayscale image of the quality control test paper includes:

[0026] Acquire an upper edge line, a lower edge line, a left edge line, and a right edge line of the first strip image in the grayscale image of the quality control test paper;

[0027] The upper edge line, the lower edge line, the left edge line and the lower edge line of the first strip image are used as dividing lines to cut out the first strip image from the grayscale image of the quality control test paper.

[0028] Optionally, the step of acquiring an upper edge line, a lower edge line, a left edge line, and a right edge line of the first strip image in the grayscale image of the quality control test paper includes:

[0029] Setting a first horizontal line calibration rectangular frame in the grayscale image of the quality control test strip, wherein the first horizontal line calibration rectangular frame covers at least a portion of the first horizontal edge of the reference line;

[0030] Obtaining a point with the largest difference between adjacent pixels in the horizontal direction within the first horizontal line calibration rectangular frame, and recording it as a first positioning point;

[0031] Connect all the first positioning points into a straight line and use it as a first calibration line;

[0032] Calculating an angle α formed between the first calibration line and the horizontal line; if α is zero, using the first calibration line as the first edge line of the first strip image; if α is not zero, flipping the grayscale image of the quality control test strip at an angle α such that the flipped first calibration line is parallel to the horizontal line, and using the flipped first calibration line as the first horizontal edge line of the first strip image;

[0033] A second horizontal line calibration rectangular frame, a first vertical line calibration rectangular frame, and a second vertical line calibration rectangular frame are set in the grayscale image of the quality control test strip, wherein the second horizontal line calibration rectangular frame covers at least a portion of the second horizontal edge of the reference line, the first vertical line calibration rectangular frame is located to the left of all the reference lines, and the second vertical line calibration rectangular frame is located to the right of all the reference lines;

[0034] Obtaining the point with the largest difference between adjacent pixels in the horizontal direction within the second horizontal line calibration rectangular frame and recording it as the second positioning point, obtaining the point with the largest difference between adjacent pixels in the vertical direction within the first vertical line calibration rectangular frame and recording it as the third positioning point, and obtaining the point with the largest difference between adjacent pixels in the vertical direction within the second vertical line calibration rectangular frame and recording it as the fourth positioning point;

[0035] Connecting all the second positioning points into a straight line as the second horizontal edge line of the first strip image, connecting all the third positioning points into a straight line as the left edge line of the first strip image, and connecting all the fourth positioning points into a straight line as the right edge line of the first strip image;

[0036] The first horizontal edge is one of the upper edge and the lower edge of the baseline, the second horizontal edge is the other of the upper edge and the lower edge of the baseline, the first horizontal edge line is one of the upper edge line and the lower edge line of the first strip image, and the second horizontal edge line is the other of the upper edge line and the lower edge line of the first strip image.

[0037] Optionally, the step of obtaining the grayscale value of each reaction line according to the standard grayscale image of the immunoblotting detection membrane strip includes:

[0038] Cutting out a second strip image including all the reaction lines from the standard grayscale image of the immunoblotting detection membrane strip;

[0039] Counting the total number of pixels of each reaction line;

[0040] Accumulating the grayscale values ​​of all pixels of each reaction line to obtain a total grayscale value of each reaction line;

[0041] The ratio of the total grayscale value of the corresponding reaction line to the total number of pixels is calculated as the grayscale value of the corresponding reaction line.

[0042] Optionally, the step of extracting a second strip image including all the reaction lines from the standard grayscale image of the immunoblotting detection membrane strip comprises:

[0043] Acquire an upper edge line, a lower edge line, a left edge line, and a right edge line of the second strip image in the standard grayscale image of the immunoblot detection membrane strip;

[0044] The upper edge line, the lower edge line, the left edge line and the lower edge line of the second strip image are used as dividing lines to cut out the second strip image from the standard grayscale image.

[0045] Optionally, the step of obtaining the upper edge line, the lower edge line, the left edge line and the lower edge line of the second strip image in the standard grayscale image of the immunoblotting detection membrane strip includes:

[0046] Setting a third horizontal line calibration rectangular frame in the standard grayscale image of the immunoblot detection membrane strip, wherein the third horizontal line calibration rectangular frame covers at least a portion of the third horizontal edge of the reaction line;

[0047] Obtaining a point with the largest difference between adjacent pixels in the horizontal direction within the third horizontal line calibration rectangular frame, and recording it as a fifth positioning point;

[0048] Connect all the fifth positioning points into a straight line, and use it as the second calibration line;

[0049] Calculating an angle ε formed between the second calibration line and the horizontal line; if ε is zero, using the second calibration line as the third horizontal edge line of the second strip image; if ε is not zero, flipping the standard grayscale image of the immunoblot detection membrane strip at a flipping angle of ε such that the flipped second calibration line is parallel to the horizontal line, and using the flipped second calibration line as the third horizontal edge line of the second strip image;

[0050] A fourth horizontal line calibration rectangular frame, a third vertical line calibration rectangular frame, and a fourth vertical line calibration rectangular frame are set in the standard grayscale image of the immunoblot detection membrane strip, wherein the fourth horizontal line calibration rectangular frame covers at least a portion of the fourth horizontal edge of the reaction line, the third vertical line calibration rectangular frame is located on the left side of the reaction line, and the fourth vertical line calibration rectangular frame is located on the right side of all the reaction lines;

[0051] Obtaining the point with the largest difference between adjacent pixels in the horizontal direction within the fourth horizontal line calibration rectangular frame and recording it as the sixth positioning point, obtaining the point with the largest difference between adjacent pixels in the vertical direction within the third vertical line calibration rectangular frame and recording it as the seventh positioning point, and obtaining the point with the largest difference between adjacent pixels in the vertical direction within the fourth vertical line calibration rectangular frame and recording it as the eighth positioning point;

[0052] connecting all the sixth positioning points into a straight line as a fourth horizontal edge line of the second strip image, connecting all the seventh positioning points into a straight line as a left edge line of the second strip image, and connecting all the eighth positioning points into a straight line as a right edge line of the second strip image;

[0053] The third horizontal edge is one of the upper edge and the lower edge of the reaction line, the fourth horizontal edge is the other of the upper edge and the lower edge of the reaction line, the third horizontal edge line is one of the upper edge line and the lower edge line of the second strip image, and the fourth horizontal edge line is the other of the upper edge line and the lower edge line of the second strip image.

[0054] Optionally, the quantitative evaluation method further includes:

[0055] Determine whether the immunoblotting detection membrane strip is valid.

[0056] Optionally, the immunoblotting detection membrane strip further has a quality control line;

[0057] The step of determining whether the immunoblotting detection membrane strip is effective comprises:

[0058] Acquiring the grayscale value of the quality control line on the standard grayscale image;

[0059] Obtaining the reflectivity of the quality control line according to the grayscale value of the quality control line and the conversion relationship between the grayscale value and the reflectivity;

[0060] The validity of the immunoblotting detection membrane strip is determined based on the reflectivity of the quality control line.

[0061] Optionally, the step of obtaining the grayscale value of the quality control line on the standard grayscale image includes:

[0062] Accumulating the grayscale values ​​of all pixels of the quality control line on the standard grayscale image to obtain a total grayscale value of the quality control line;

[0063] Counting the total number of pixels of the quality control line;

[0064] The ratio of the total grayscale value of the quality control line to the total number of pixels of the quality control line is calculated as the grayscale value of the quality control line.

[0065] To achieve the above object, the present invention further provides a computer-readable storage medium having a program stored thereon, which, when executed, performs the quantitative evaluation method for immunoblotting detection membrane strips as described above.

[0066] To achieve the above object, the present invention further provides an electronic device, comprising a processor and the computer-readable storage medium as described above, wherein the processor is configured to execute a program stored on the computer-readable storage medium.

[0067] To achieve the above-mentioned purpose, the present invention also provides a quantitative evaluation device for immunoblot detection membrane strips, comprising a camera and a control unit, wherein the camera is used to take pictures of the immunoblot detection membrane strip to obtain a standard color image of the immunoblot detection membrane strip, and the control unit is communicatively connected to the camera and is configured to execute the quantitative evaluation method for the immunoblot detection membrane strip as described above.

[0068] Compared with the prior art, the immunoblotting detection membrane strip evaluation method and device, storage medium and electronic device of the present invention have the following advantages:

[0069] The immunoblot detection membrane strip has multiple reaction lines; the quantitative evaluation method of the immunoblot detection membrane strip includes the following steps: obtaining a standard color image of the immunoblot detection membrane strip; converting the standard color image of the immunoblot detection membrane strip into a standard grayscale image; obtaining the grayscale value of each of the reaction lines in the standard grayscale image; obtaining the reflectivity of each of the reaction lines based on the grayscale value of each of the reaction lines and the pre-set conversion relationship between the grayscale value and the reflectivity; quantitatively evaluating the characteristics of the protein detected by the immunoblot detection membrane strip based on the reflectivity of each of the reaction lines. This method uses reflectivity to achieve quantitative analysis of the immunoblot detection results, and has the advantages of being empirical, clear, objective, and traceable. When this quantitative evaluation method is applied to the medical field, it can help doctors give more reasonable, more effective, and more accurate treatment plans based on accurate quantitative results, improve treatment effects, and also achieve the purpose of traceability of medical data. BRIEF DESCRIPTION OF THE DRAWINGS

[0070] The accompanying drawings are provided for a better understanding of the present invention and are not intended to limit the present invention.

[0071] FIG1 is a flow chart of a quantitative evaluation method for an improved immunoblotting detection membrane strip according to one embodiment of the present invention;

[0072] FIG2 is a schematic diagram of a quality control test paper used in a quantitative evaluation method for an immunoblot detection membrane strip according to an embodiment of the present invention.

[0073] 3 is a curve showing the relationship between the grayscale value and reflectivity of a quality control strip of a quality control test paper used in a quantitative evaluation method for an immunoblot detection membrane strip according to one embodiment of the present invention;

[0074] FIG4 is a flowchart of the steps of setting up a standard shooting environment in a quantitative evaluation method for immunoblotting detection membrane strips according to one embodiment of the present invention;

[0075] 5 is a schematic diagram of setting a first horizontal line calibration rectangular frame, a second horizontal line calibration rectangular frame, a first vertical line calibration rectangular frame, and a second vertical line calibration rectangular frame in a grayscale image of a quality control test strip when obtaining a standard shooting environment in a quantitative evaluation method for an immunoblot detection membrane strip provided according to one embodiment of the present invention;

[0076] FIG6 is a schematic diagram of a first upper edge line, a first lower edge line, a first left edge line, and a first right edge line obtained in a second grayscale image of a quality control test strip when a standard shooting environment is obtained in a quantitative evaluation method for an immunoblot detection membrane strip provided according to an embodiment of the present invention;

[0077] 7 is a schematic diagram of setting a third horizontal line calibration rectangular frame, a fourth horizontal line calibration rectangular frame, a third vertical line calibration rectangular frame, a fourth vertical line calibration rectangular frame, and a fifth vertical line calibration rectangular frame in a standard grayscale image of an immunoblot detection membrane strip in a quantitative evaluation method of an immunoblot detection membrane strip provided according to one embodiment of the present invention;

[0078] Figure 8 is a schematic diagram of the second upper edge line, second lower edge line, second left edge line, second right edge line and quality control line calibration line obtained from the standard grayscale image of the immunoblot detection membrane strip in the quantitative evaluation method of the immunoblot detection membrane strip provided according to one embodiment of the present invention.

[0079] In the accompanying drawings: 1-baseline, 2-first upper edge line, 3-first lower edge line, 4-first left edge line, 5-first right edge line, 6-first horizontal line calibration rectangular box, 7-second horizontal line calibration rectangular box, 8-first vertical line calibration rectangular box, 9-second vertical line calibration rectangular box; 10-reaction line, 20-second upper edge line, 30-second lower edge line, 40-second left edge line, 50-second right edge line, 60-third horizontal line calibration rectangular box, 70-fourth horizontal line calibration rectangular box, 80-third vertical line calibration rectangular box, 90-fourth vertical line calibration rectangular box; 100-quality control line. DETAILED DESCRIPTION

[0080] The following describes the embodiments of the present invention through specific specific examples. Those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the illustrations provided in this embodiment only illustrate the basic concept of the present invention in a schematic manner, and the drawings only show components related to the present invention rather than being drawn according to the number, shape and size of components during actual implementation. During actual implementation, the type, quantity and proportion of each component can be changed at will, and the component layout type may also be more complicated.

[0081] In addition, each embodiment described below has one or more technical features. However, this does not mean that users of the present invention must implement all technical features in any embodiment at the same time, or that they can only implement some or all technical features in different embodiments separately. In other words, as long as implementation is possible, those skilled in the art can, based on the disclosure of the present invention and depending on design specifications or implementation requirements, selectively implement some or all technical features in any embodiment, or selectively implement a combination of some or all technical features in multiple embodiments, thereby increasing the flexibility of the implementation of the present invention.

[0082] As used in this specification, the singular forms "a," "an," and "the" include plural referents, and the plural form "a plurality" includes more than two referents, unless the context clearly indicates otherwise. As used in this specification, the term "or" is generally used in a sense including "and / or," unless the context clearly indicates otherwise. Those skilled in the art will understand the specific meanings of these terms in the present invention based on specific circumstances.

[0083] The present invention aims to provide a quantitative evaluation method for immunoblot membrane strips, aiming to automatically generate objective, accurate, and traceable quantitative analysis results for immunoblot membrane strips. These quantitative analysis results are particularly useful in the medical field, helping doctors develop targeted treatment plans for patients based on the quantitative analysis results, thereby improving treatment outcomes. Furthermore, these methods meet current regulatory requirements for traceability of medical data.

[0084] To make the objects, advantages, and features of the present invention more apparent, the present invention is further described below in detail with reference to the accompanying drawings. It should be noted that the drawings are greatly simplified and not to exact scale, and are intended solely to facilitate and clearly illustrate the embodiments of the present invention. The same or similar reference numerals in the drawings represent the same or similar components.

[0085] Those skilled in the art will appreciate that the immunoblot membrane strip to be analyzed has multiple parallel reaction lines 10. The quantitative evaluation method for the immunoblot membrane strip provided in the embodiments of the present invention quantitatively determines the properties of the detected protein by obtaining the reflectivity of the multiple reaction lines 10, as shown in FIG1 .

[0086] Referring to Figure 1, the quantitative evaluation method includes steps S100, S200, S300, S400, and S500. Specifically, step S100 includes obtaining a standard color image of the immunoblot detection membrane strip. Step S200 includes converting the standard color image of the immunoblot detection membrane strip into a standard grayscale image. Step S300 includes obtaining the grayscale value of each of the reaction lines 10 in the standard grayscale image. Step S400 includes obtaining the reflectivity of each of the reaction lines 10 based on the grayscale value of each of the reaction lines 10 in the standard grayscale image and a pre-set conversion relationship between the grayscale value and the reflectivity. Step S500 includes quantitatively evaluating the properties of the protein detected by the immunoblot detection membrane strip based on the reflectivity of each of the reaction lines.

[0087] The standard color image of the immunoblot test strip refers to a color image of the immunoblot test strip captured under a standard shooting environment. The standard shooting environment means that the intensity of light emitted by the light source used for shooting, the angle of incidence of the light, and the exposure time of the camera all meet preset requirements. The process of establishing the standard shooting environment will be described later.

[0088] The conversion relationship between the grayscale value and the reflectivity is obtained by a quality control test paper. The quality control test paper can be directly purchased from the market or designed by oneself. In the embodiment of the present invention, the quality control test paper designed by oneself is used.

[0089] Specifically, first, a rectangular frame with a length and width of Xmm*Ymm is drawn using drawing software. The specific values ​​of X and Y are determined as needed. The embodiment of the present invention does not limit this, but X is usually greater than Y. Then, n reference lines 1 are set at equal intervals in the rectangular frame (as shown in Figure 2), where n is a positive integer greater than 1, and the specific value is set as needed. Then, each of the reference lines 1 is grayscale filled in turn to obtain the quality control test paper shown in Figure 2. The filling principle is: the R value, G value and B value of each reference line 1 are the same, and the difference in grayscale values ​​between two adjacent reference lines 1 is 255 / (n-1). That is, the R value, G value and B value of the i-th reference line 1 are all set to 255×(i-1) / (n-1). It should be noted that the R value, G value and B value of each reference line 1 are all integers. If the result calculated according to 255×(i-1) / (n-1) is not an integer, an integer is obtained according to the principle of rounding. Table 1 shows the grayscale values ​​of n reference lines 1 .

[0090] Table 1

[0091] Next, the quality control test strips were printed using a high-resolution color photocopying device on matte-surface white photo paper. The reflectivity of each baseline 1 on the printed quality control test strips was then measured using a reflectivity meter. The results are shown in Table 2. The measurement standard was JJG453-2002, "Standard Color Plate Verification Procedure."

[0092] Table 2

[0093] According to the gray value of the baseline 1 in the quality control test paper and its corresponding reflectivity, a curve as shown in FIG3 is obtained. Finally, the curve is polynomially fitted using MATLAB software and the Polynomial Regression function, and the conversion relationship between the gray value and the reflectivity is in accordance with formula (1). Formula (1) is: R = A1 * G V +A2*G V 2+A3*G V 3 +……A n *G V n ,

[0094] In the formula, R represents reflectivity, Gv represents grayscale value, A n Represents the polynomial coefficients.

[0095] The standard shooting environment is constructed based on the quality control test paper, and the operation of constructing the standard shooting environment is performed before step S100. The specific construction process is as follows:

[0096] First, hardware is provided, including a camera for taking pictures and a light source for lighting. Next, the construction operation shown in Figure 4 is performed. As shown in Figure 4, the construction operation includes the following steps S10, S20, S30, S40 and S50. Step S10 includes taking pictures of the quality control test paper using the camera to obtain a color image of the quality control test paper. Step S20 includes converting the color image of the quality control test paper into a grayscale image of the quality control test paper. Step S30 includes obtaining the grayscale value of each of the baselines on the grayscale image of the quality control test paper. Step S40 includes determining whether the grayscale value of each baseline is the same as the corresponding set grayscale value. If so, it is considered that the current shooting environment is the standard shooting environment. If not, step S50 is executed. Step S50 includes adjusting at least one of the intensity of the light emitted by the light source, the incident angle of the light and the exposure time of the camera.

[0097] It is understood that steps S10 to S40 are performed at least once. After step S50 is completed, the process returns to step S10 to S40. In other words, the standard shooting environment refers to the shooting environment when the grayscale value of each baseline 1 in the grayscale image of the quality control test paper corresponding to the color image of the quality control test paper obtained by photographing the quality control test paper is equal to the corresponding set grayscale value.

[0098] It can also be understood that the steps S20 to S40 are performed by a control unit such as a computer. Preferably, the control unit is also connected to the camera and the light source, and preferably the control unit controls the camera to take pictures of the immunoblotting detection film strip, and controls the exposure time of the camera, and the control unit also controls the intensity of the light emitted by the light source. Further, the light source is connected to a posture adjustment mechanism, and the control unit is also connected to the posture adjustment mechanism, and adjusts the posture of the light source by controlling the movement of the posture adjustment mechanism to adjust the angle of incidence of the light emitted by the light source. That is, the steps S10 and S50 can be performed under the control of the control unit.

[0099] Optionally, the step S30 is performed according to the following steps S31, S32 and S33 (not shown in the figure). In detail, the step S31 includes cutting out a first strip image including all the baselines 1 in the grayscale image of the quality control test paper. The step S32 includes counting the total number of pixels of each baseline 1 in the first strip image, and adding up the grayscale values ​​of all pixels of each baseline 1 to obtain the total grayscale value of each baseline 1. The step S33 includes dividing the total grayscale value of any baseline 1 by the total number of pixels of the baseline 1, and using the obtained quotient as the grayscale value of the corresponding baseline 1. Those skilled in the art know that the color image of the quality control test paper obtained by the step S10 includes not only the image information of all the baselines 1, but also the external environment image information outside the baseline 1. The presence of the external environment image information may affect the analysis of the baseline 1 and increase the amount of calculation when the control unit processes the image. Therefore, by extracting the first strip image and obtaining the grayscale value of each baseline 1 based on the first strip image, on the one hand, there is no need to perform data processing on the external environment image information in the grayscale image of the quality control test strip, which reduces the computational effort of the control unit and the configuration requirements for the control unit. On the other hand, it also avoids interference with the grayscale value of the baseline 1 caused by the external environment image information. It is worth noting that steps S32 and S33 are executed for each baseline 1, and therefore the number of executions of steps S32 and S33 is the same as the number of baselines 1, that is, steps S32 and S33 are each executed n times.

[0100] In more detail, the upper edge line, lower edge line, left edge line and right edge line of the first strip image can be obtained on the grayscale image of the quality control test paper, and then the upper edge line, lower edge line, left edge line and right edge line of the first strip image can be used as dividing lines to segment and cut out the first strip image from the grayscale image of the quality control test paper.

[0101] As mentioned above, all the reference lines 1 are designed to be within a rectangular frame with a length and width of Xmm*Ymm, so the first strip image is usually in a rectangular shape. In this article, the extension direction of the two long sides of the rectangular shape is taken as the horizontal direction, the extension direction of the two short sides is taken as the vertical direction, and the two long sides of the first strip image are respectively referred to as the upper edge line and the lower edge line, and the two short sides are respectively referred to as the left edge line and the right edge line. In order to facilitate the distinction from the upper edge line, lower edge line, left edge line and right edge line of the immunoblotting detection membrane strip image described later, the upper edge line of the first strip image is referred to as the first upper edge line 2, the lower edge line of the first strip image is referred to as the first lower edge line 3, the left edge line of the first strip image is referred to as the first left edge line 4, and the right edge line of the first strip image is referred to as the first right edge line 5.

[0102] The specific steps of obtaining the first upper edge line 2, the first lower edge line 3, the first left edge line 4, and the first right edge line 5 in the grayscale image of the quality control test paper are as follows:

[0103] First, as shown in FIG5 , a first horizontal line calibration rectangular frame 6 is set in the grayscale image of the quality control test paper, and the first horizontal line calibration rectangular frame 6 covers at least a portion of the first horizontal edge of the reference line 1 .

[0104] Then, the point with the largest difference between adjacent pixels in the horizontal direction within the first horizontal line calibration rectangular frame 6 is obtained and recorded as the first positioning point. In practice, multiple first positioning points can be obtained by executing this step.

[0105] Then, all the first positioning points are connected into a straight line, which is recorded as a first marking line.

[0106] Then, the angle α formed between the first calibration line and the horizontal line is calculated. If α is zero, the first calibration line is used as the first horizontal edge line of the first strip image. If α is not equal to zero, the grayscale image of the quality control test strip is flipped at an angle of α, such that the flipped first calibration line is parallel to the horizontal line. The flipped first calibration line is used as the first horizontal edge line of the first strip image.

[0107] Next, as shown in FIG5 , a second horizontal line calibration rectangular frame 7, a first vertical line calibration rectangular frame 8, and a second vertical line calibration rectangular frame 9 are set in the grayscale image of the quality control test strip. The second horizontal line calibration rectangular frame 7 covers at least a portion of the second horizontal edge of the reference line 1, the first vertical line calibration rectangular frame 8 is located on the left side of all the reference lines 1, and the second vertical line calibration rectangular frame 9 is located on the right side of all the reference lines 1.

[0108] Next, the point with the largest difference between adjacent pixels in the horizontal direction within the second horizontal line calibration rectangular frame 7 is obtained and recorded as the second positioning point. The point with the largest difference between adjacent pixels in the vertical direction within the first vertical line calibration rectangular frame 8 is obtained and recorded as the third positioning point. The point with the largest difference between adjacent pixels in the vertical direction within the second vertical line calibration rectangular frame 9 is obtained and recorded as the fourth positioning point. Through this step, multiple second positioning points, multiple third positioning points, and multiple fourth positioning points are obtained.

[0109] Then, all the second positioning points are connected into a straight line as the second horizontal edge line of the first strip image, all the third positioning points are connected into a straight line as the first left edge line 4 (as shown in Figure 6), and all the fourth positioning points are connected into a straight line as the first right edge line 5 (as shown in Figure 6).

[0110] In an embodiment of the present invention, the first horizontal edge is one of the upper edge and the lower edge of the reference line 1, and the second horizontal edge is the other of the upper edge and the lower edge of the reference line 1. Similarly, the first horizontal edge line is one of the first upper edge line 2 and the first lower edge line 3, and the second horizontal edge line is the other of the first upper edge line 2 and the first lower edge line 3. In a specific implementation, the first horizontal edge is the upper edge of the reference line 1, the second horizontal edge is the lower edge of the reference line 1, and the first horizontal edge line is the first upper edge line 2, and the second horizontal edge line is the first lower edge line 3.

[0111] After the standard shooting environment is built, the various steps in the evaluation method of the immunoblotting detection film strip can be performed. Wherein, the operation of the step S300 is substantially the same as the operation of the step S30. That is to say, the step S300 may specifically include the following steps S310, step S320 and step S330 (not shown in the figure). The step S310 includes cutting out a second strip image including all reaction lines 10 in the standard grayscale image of the immunoblotting detection film strip. The step S320 includes counting the total number of pixels of each of the reaction lines 10 in the second strip image, and adding up the grayscale values ​​of all pixels of each of the reaction lines 10 to obtain the total grayscale value of each of the reaction lines 10. The step S330 includes dividing the total grayscale value of any one of the reaction lines 10 by the corresponding total number of pixels, and using the obtained quotient as the grayscale value of the corresponding reaction line 10. Steps S320 and S330 are performed for each reactive line 10. Therefore, the number of executions of steps S320 and S330 is equal to the number of reactive lines 10. After obtaining the grayscale value of each reactive line 10, the grayscale value of each reactive line 10 is substituted into formula (1) to obtain the reflectivity of each reactive line 10.

[0112] The step of cutting out the second strip image from the standard grayscale image of the immunoblot detection membrane strip includes: first obtaining the upper edge line, lower edge line, left edge line, and right edge line of the second strip image from the standard grayscale image of the immunoblot detection membrane strip, and then using the upper edge line, lower edge line, left edge line, and right edge line of the second strip image as dividing lines to segment and cut out the second strip image. Hereinafter, the upper edge line of the second strip image will be referred to as the second upper edge line 20 (as shown in FIG8 ), the lower edge line of the second strip image will be referred to as the second lower edge line 30, the left edge line of the second strip image will be referred to as the second left edge line 40, and the right edge line of the second strip image will be referred to as the second right edge line 50.

[0113] The specific steps of obtaining the second upper edge line 20, the second lower edge line 30, the second left edge line 40 and the second right edge line 50 in the standard grayscale image of the immunoblot detection membrane strip are as follows:

[0114] First, as shown in FIG7 , a third horizontal line calibration rectangular frame 60 is set in the grayscale image of the quality control test strip, and the third horizontal line calibration rectangular frame 60 covers at least a portion of the third horizontal edge of the reaction line 10 .

[0115] Then, the point with the largest difference between adjacent pixels in the horizontal direction within the third horizontal line calibration rectangular frame 60 is obtained and recorded as the fifth positioning point. In this step, multiple fifth calibration points can be obtained.

[0116] Then, all the fifth positioning points are connected into a straight line, which is recorded as a second marking line.

[0117] Then, the angle ε formed between the second calibration line and the horizontal line is calculated. If ε is zero, the second calibration line is used as the third horizontal edge line of the second strip image. If ε is not zero, the standard grayscale image of the immunoblot detection membrane strip is flipped at an angle of ε, such that the flipped second calibration line is parallel to the horizontal line. The flipped second calibration line is used as the third horizontal edge line of the second strip image.

[0118] Next, as shown in FIG7 , a fourth horizontal line marking rectangular frame 70, a third vertical line marking rectangular frame 80, and a fourth vertical line marking rectangular frame 90 are set in the standard grayscale image of the immunoblot detection membrane strip. The fourth horizontal line marking rectangular frame 70 covers at least a portion of the fourth horizontal edge of the reaction lines 10, the third vertical line marking rectangular frame 80 is located on the left side of all the reaction lines 10, and the fourth vertical line marking rectangular frame 90 is located on the right side of all the reaction lines 10.

[0119] Next, the point with the largest difference between adjacent pixels in the horizontal direction within the fourth horizontal line calibration rectangular frame 70 is obtained and recorded as the sixth positioning point. The point with the largest difference between adjacent pixels in the vertical direction within the third vertical line calibration rectangular frame 80 is obtained and recorded as the seventh positioning point. The point with the largest difference between adjacent pixels in the vertical direction within the fourth vertical line calibration rectangular frame 90 is obtained and recorded as the eighth positioning point. Through this step, multiple sixth positioning points, multiple seventh positioning points, and multiple eighth positioning points are obtained.

[0120] Finally, all the sixth positioning points are connected into a straight line as the fourth horizontal edge line of the second strip image, all the seventh positioning points are connected into a straight line as the second left edge line 40 (as shown in Figure 8), and all the eighth positioning points are connected into a straight line as the second right edge line 50 (as shown in Figure 8).

[0121] In the embodiment of the present invention, the third horizontal edge is one of the upper edge and the lower edge of the reaction line 10, and the fourth horizontal edge is the other of the upper edge and the lower edge of the reaction line 10. Similarly, the third horizontal edge line is one of the second upper edge line 20 and the second lower edge line 30, and the fourth horizontal edge line is the other of the second upper edge line 20 and the second lower edge line 30. In a specific implementation, the third horizontal edge is the upper edge of the reaction line 10, the fourth horizontal edge is the lower edge of the reaction line 10, and the third horizontal edge line is the second upper edge line 20, and the fourth horizontal edge line is the second lower edge line 30.

[0122] In practice, the immunoblot detection membrane strip often becomes ineffective due to various reasons, such as improper operation during immunoincubation, and cannot be used to accurately determine the properties of the detected protein. In view of this, the evaluation method also includes step S600 (not shown): determining whether the immunoblot detection membrane strip is valid. Step S500 is preferably performed after step S310 and before step S320.

[0123] Optionally, the immunoblot detection membrane strip further has a quality control line 100. The step of judging whether the immunoblot detection membrane strip is valid specifically includes step S610, step S620 and step S630 (not shown in the figure). Step S610 includes obtaining the grayscale value of the quality control line 100 on the standard grayscale image. Step S620 includes obtaining the reflectivity of the quality control line 100 based on the grayscale value of the quality control line and formula (1). Step S630 includes judging whether the immunoblot detection membrane strip is valid based on the reflectivity of the quality control line 100.

[0124] The steps for obtaining the grayscale value of the quality control line 100 on the standard grayscale image are substantially the same as the steps for obtaining the grayscale value of each reaction line 10, specifically comprising: counting the total number of pixels of the quality control line; accumulating the grayscale values ​​of all pixels of the quality control line 100 on the standard grayscale image to obtain a total grayscale value of the quality control line; and then dividing the total grayscale value of the quality control line 100 by the total number of pixels of the quality control line 100, and using the resulting quotient as the grayscale value of the quality control line 100.

[0125] Next, the accuracy of the quantitative evaluation method for the immunoblotting detection membrane strip provided in the embodiment of the present invention is introduced.

[0126] First, a quantitative-semi-quantitative-qualitative comparison table of reflectance was prepared based on a large number of clinical test samples, as shown in Table 3 below.

[0127] Table 3

[0128] Next, an inactivated, 20-item immunoblot test strip with multiple reaction results was selected. The allergens on it are shown in Table 4. It should be understood that in this embodiment, each allergen line is a reaction line 10. Manual interpretation, semi-quantitative evaluation using the Mediwiss Allergen Screen C1 instrument, and quantitative evaluation using the quantitative evaluation method provided in an embodiment of the present invention were then performed. The results are shown in Table 4. In this embodiment, the reflectivity of the quality control line 100 was set to less than 0.805, indicating that the immunoblot test strip was valid when the quality control line was positive.

[0129] Table 4

[0130] As can be seen from Table 4, the quantitative analysis results of the embodiment of the present invention are the same as the manual interpretation results and the detection results of the Mediwiss sensitive screening allergen detector C1.

[0131] In summary, the quantitative evaluation method for immunoblotting detection membrane strips provided in the embodiments of the present invention uses the reflectance of the reaction line as the quantitative parameter, which not only automatically, objectively and accurately obtains the analysis results, but also has the advantage of traceability of the results.

[0132] Furthermore, an embodiment of the present invention also provides a computer-readable storage medium having a program stored thereon. When the program is executed, the steps in the aforementioned quantitative evaluation method for immunoblotting detection membrane strips are performed.

[0133] Furthermore, an embodiment of the present invention further provides an electronic device, which includes a processor and the computer-readable storage medium, and the processor is configured to execute a program stored on the computer-readable storage medium.

[0134] Furthermore, an embodiment of the present invention also provides an evaluation device for an immunoblot detection membrane strip, the evaluation device comprising a camera and a control unit, the camera being used to take a picture of the immunoblot detection membrane strip to obtain a standard color image of the immunoblot detection membrane strip, the control unit being communicatively connected to the camera and configured to execute the quantitative evaluation method for the immunoblot detection membrane strip as described above.

[0135] Preferably, the control unit is further configured to control the camera to take pictures of the immunoblotting detection membrane strip and control the exposure time of the camera.

[0136] Preferably, the evaluation device further comprises a light source and a position adjustment mechanism connected to the light source. The control unit is communicatively connected to the light source and the position adjustment mechanism, and is configured to control the intensity of the light generated by the light source and control the movement of the position adjustment mechanism to adjust the position of the light source, thereby adjusting the incident angle of the light emitted by the light source.

[0137] While the present invention is disclosed above, it is not limited thereto. Those skilled in the art may make various modifications and variations to the present invention without departing from the spirit and scope of the present invention. Thus, the present invention is intended to encompass such modifications and variations as long as they fall within the scope of the claims and their equivalents.

Claims

1. A quantitative evaluation method for an immunoblot detection membrane strip, wherein the immunoblot detection membrane strip has a plurality of reaction lines, It is characterized in that The evaluation method comprises the following steps: Obtaining a standard color image of the immunoblotting detection membrane strip; Converting the standard color image of the immunoblot detection membrane strip into a standard grayscale image; Acquire the grayscale value of each of the reaction lines in the standard grayscale image; Obtaining the reflectivity of each of the reaction lines according to the grayscale value of each of the reaction lines in the standard grayscale image and a preset conversion relationship between the grayscale value and the reflectivity; The characteristics of the protein detected by the immunoblotting detection membrane strip are quantitatively evaluated according to the reflectance of each reaction line.

2. The quantitative evaluation method of the immunoblotting detection membrane strip according to claim 1, It is characterized in that The conversion relationship between grayscale value and reflectivity is obtained by setting grayscale values ​​of multiple baselines on the quality control test paper and corresponding reflectivity; And, the standard color image is obtained by photographing the immunoblot detection membrane strip under a standard shooting environment; the standard shooting environment refers to a shooting environment in which the quality control test paper is photographed to obtain a quality control test paper color image, and the grayscale value of each baseline in the quality control test paper grayscale image corresponding to the quality control test paper color image is equal to the corresponding set grayscale value.

3. The quantitative evaluation method of the immunoblotting detection membrane strip according to claim 2, It is characterized in that The step of obtaining the standard shooting environment includes: Taking a photo of the quality control test paper to obtain a color image of the quality control test paper; Converting the quality control test paper color image into a quality control test paper grayscale image; Obtaining the grayscale value of each of the baselines on the grayscale image of the quality control test paper; Determine whether the grayscale value of each baseline on the grayscale image of the quality control test paper is equal to the corresponding set grayscale value. If so, use the current shooting environment as the standard shooting environment. If not, adjust at least one of the intensity of the light emitted by the light source used for lighting, the incident angle of the light, and the exposure time of the camera.

4. The quantitative evaluation method of the immunoblotting detection membrane strip according to claim 3, It is characterized in that The step of obtaining the grayscale value of each baseline on the grayscale image of the quality control test paper comprises: Cutting out a first strip image including all the reference lines from the grayscale image of the quality control test paper; Counting the total number of pixels of each of the baselines; Accumulating the grayscale values ​​of all pixels of each of the reference lines to obtain a total grayscale value of each of the reference lines; The ratio of the total grayscale value of the corresponding reference line to the total number of pixels is calculated as the grayscale value of the corresponding reference line.

5. The quantitative evaluation method of the immunoblotting detection membrane strip according to claim 4, It is characterized in that The step of cutting out a first strip image including all the reference lines in the grayscale image of the quality control test paper comprises: Acquire an upper edge line, a lower edge line, a left edge line, and a right edge line of the first strip image in the grayscale image of the quality control test paper; The upper edge line, the lower edge line, the left edge line and the lower edge line of the first strip image are used as segmentation lines to cut out the first strip image in the grayscale image of the quality control test paper.

6. The quantitative evaluation method of the immunoblotting detection membrane strip according to claim 5, It is characterized in that The step of obtaining the upper edge line, the lower edge line, the left edge line and the right edge line of the first strip image in the grayscale image of the quality control test paper comprises: Setting a first horizontal line calibration rectangular frame in the grayscale image of the quality control test paper, wherein the first horizontal line calibration rectangular frame covers at least a portion of the first horizontal edge of the reference line; Obtaining a point with the largest adjacent pixel difference in the horizontal direction within the first horizontal line calibration rectangular frame, and recording it as a first positioning point; Connect all the first positioning points into a straight line as a first calibration line; Calculate the angle α formed by the first calibration line and the horizontal line. If α is zero, use the first calibration line as the first edge line of the first strip image. If α is not equal to zero, flip the grayscale image of the quality control test paper. The flip angle is α, so that the first calibration line after flipping is parallel to the horizontal line, and use the first calibration line after flipping as the first horizontal edge line of the first strip image. A second horizontal line calibration rectangular frame, a first vertical line calibration rectangular frame and a second vertical line calibration rectangular frame are set in the grayscale image of the quality control test paper, and the second horizontal line calibration rectangular frame covers at least part of the The second horizontal edge of the reference lines is divided, the first vertical line marking rectangular frame is located on the left side of all the reference lines, and the second vertical line marking rectangular frame is located on the right side of all the reference lines; Obtaining the point with the largest difference between adjacent pixels in the horizontal direction in the second horizontal line calibration rectangular frame and recording it as the second positioning point, obtaining the point with the largest difference between adjacent pixels in the vertical direction in the first vertical line calibration rectangular frame and recording it as the third positioning point, and obtaining the point with the largest difference between adjacent pixels in the vertical direction in the second vertical line calibration rectangular frame and recording it as the fourth positioning point; Connecting all the second positioning points into a straight line as a second horizontal edge line of the first strip image, connecting all the third positioning points into a straight line as a left edge line of the first strip image, and connecting all the fourth positioning points into a straight line as a right edge line of the first strip image; The first horizontal edge is one of the upper edge and the lower edge of the baseline, the second horizontal edge is the other of the upper edge and the lower edge of the baseline, the first horizontal edge line is one of the upper edge line and the lower edge line of the first strip image, and the second horizontal edge line is the other of the upper edge line and the lower edge line of the first strip image.

7. The quantitative evaluation method of the immunoblotting detection membrane strip according to claim 1, It is characterized in that The step of obtaining the grayscale value of each of the reaction lines according to the standard grayscale image of the immunoblotting detection membrane strip comprises: Cutting out a second strip image including all the reaction lines from the standard grayscale image of the immunoblot detection membrane strip; Counting the total number of pixels of each reaction line; Accumulating the grayscale values ​​of all pixels of each of the reaction lines to obtain a total grayscale value of each of the reaction lines; The ratio of the total grayscale value of the corresponding reaction line to the total number of pixels is calculated as the grayscale value of the corresponding reaction line.

8. The quantitative evaluation method of the immunoblotting detection membrane strip according to claim 7, It is characterized in that The step of cutting out a second strip image including all the reaction lines from the standard grayscale image of the immunoblotting detection membrane strip comprises: Acquire the upper edge line, the lower edge line, the left edge line and the right edge line of the second strip image in the standard grayscale image of the immunoblot detection membrane strip; The upper edge line, the lower edge line, the left edge line and the lower edge line of the second strip image are used as segmentation lines to cut out the second strip image from the standard grayscale image.

9. The quantitative evaluation method of the immunoblotting detection membrane strip according to claim 8, It is characterized in that The step of acquiring the upper edge line, the lower edge line, the left edge line and the lower edge line of the second strip image in the standard grayscale image of the immunoblot detection membrane strip comprises: Setting a third horizontal line calibration rectangular frame in the standard grayscale image of the immunoblot detection membrane strip, wherein the third horizontal line calibration rectangular frame covers at least a portion of the third horizontal edge line of the reaction line; Obtaining a point with the largest adjacent pixel difference in the horizontal direction within the third horizontal line calibration rectangular frame, and recording it as a fifth positioning point; Connect all the fifth positioning points into a straight line as a second calibration line; Calculate the angle ε formed by the second calibration line and the horizontal line; if ε is zero, use the second calibration line as the third horizontal edge line of the second strip image; if ε is not equal to zero, flip the standard grayscale image of the immunoblot detection membrane strip, and the flip angle is ε, so that the flipped second calibration line is parallel to the horizontal line, and use the flipped second calibration line as the third horizontal edge line of the second strip image; A fourth horizontal line calibration rectangular frame, a third vertical line calibration rectangular frame and a fourth vertical line calibration rectangular frame are set in the standard grayscale image of the immunoblot detection membrane strip, wherein the fourth horizontal line calibration rectangular frame covers at least a portion of the fourth horizontal edge of the reaction line, the third vertical line calibration rectangular frame is located on the left side of the reaction line, and the fourth vertical line calibration rectangular frame is located on the right side of all the reaction lines; Obtain the point with the largest difference between adjacent pixels in the horizontal direction in the fourth horizontal line calibration rectangular frame and record it as the sixth positioning point, obtain the point with the largest difference between adjacent pixels in the vertical direction in the third vertical line calibration rectangular frame and record it as the seventh positioning point, and obtain the point with the largest difference between adjacent pixels in the vertical direction in the fourth vertical line calibration rectangular frame and record it as the eighth positioning point; Connecting all the sixth positioning points into a straight line as a fourth horizontal edge line of the second strip image, connecting all the seventh positioning points into a straight line as a left edge line of the second strip image, and connecting all the eighth positioning points into a straight line as a right edge line of the second strip image; The third horizontal edge is one of the upper edge and the lower edge of the reaction line. The horizontal edge is the other of the upper edge and the lower edge of the reaction line, the third horizontal edge line is one of the upper edge line and the lower edge line of the second strip image, and the fourth horizontal edge line is the other of the upper edge line and the lower edge line of the second strip image.

10. The quantitative evaluation method of the immunoblotting detection membrane strip according to claim 9, It is characterized in that The quantitative evaluation method also includes: Determine whether the immunoblotting detection membrane strip is effective.

11. The quantitative evaluation method of the immunoblotting detection membrane strip according to claim 10, It is characterized in that The immunoblotting detection membrane strip also has a quality control line; The step of judging whether the immunoblotting detection membrane strip is effective comprises: Acquiring the grayscale value of the quality control line on the standard grayscale image; Obtaining the reflectivity of the quality control line according to the gray value of the quality control line and the conversion relationship between the gray value and the reflectivity; Whether the immunoblotting detection membrane strip is valid is judged according to the reflectivity of the quality control line.

12. The quantitative evaluation method of the immunoblotting detection membrane strip according to claim 11, It is characterized in that The step of obtaining the grayscale value of the quality control line on the standard grayscale image comprises: Accumulating the grayscale values ​​of all pixels of the quality control line on the standard grayscale image to obtain the total grayscale value of the quality control line; Counting the total number of pixels of the quality control line; The ratio of the total grayscale value of the quality control line to the total number of pixels of the quality control line is calculated as the grayscale value of the quality control line.

13. A computer-readable storage medium having a program stored thereon, It is characterized in that When the program is executed, the quantitative evaluation method of the immunoblotting detection membrane strip according to any one of claims 1 to 12 is performed.

14. An electronic device, It is characterized in that The method comprises a processor and the computer-readable storage medium according to claim 13, wherein the processor is configured to execute a program stored on the computer-readable storage medium.

15. A quantitative evaluation device for immunoblotting detection membrane strips, It is characterized in that It comprises a camera and a control unit, wherein the camera is used to take a picture of the immunoblot detection membrane strip to obtain a standard color image of the immunoblot detection membrane strip, and the control unit is communicatively connected with the camera and configured to execute the quantitative evaluation method of the immunoblot detection membrane strip as described in any one of claims 1 to 12.

Images