A test strip for rapid detection of microorganisms and its preparation method

Abstract

The present invention relates to the technical field of microbial test strip preparation, and particularly to a test strip for rapidly detecting microorganisms and a preparation method thereof. In the present invention, a conjugate pad for fixing colored microspheres is pasted to one end of a nitrocellulose membrane, and the pasted conjugate pad is rolled to obtain contour images of a plurality of overlapping regions. The resilience coefficient is determined according to the comparison of contour images of different frames to determine whether there is a risk of abnormal fitting. According to the contour images of characteristic frames within a preset acquisition period, the fitting characterization parameters are determined to determine the type of abnormal spraying in the overlapping region. According to the type of abnormal spraying, the spraying adjustment method of colored microspheres is selected. A sample pad and a water absorption pad are connected to the nitrocellulose membrane and the conjugate pad to prepare a target test strip. Furthermore, the risk tendency of abnormal fitting of the nitrocellulose membrane is rapidly identified, the spraying method of colored microspheres is adaptively adjusted, and the reliability of the preparation of the test strip is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of microbial test strip preparation, and particularly to a test strip for rapid detection of microorganisms and a preparation method thereof. Background Art

[0002] Genital tract infection has always been an issue concerning reproductive health at home and abroad. It is an inflammation caused by changes in the microbial environment flora and the biochemical properties of secretions. The essence of genital tract infection is microecological imbalance. At present, the Nugent scoring method is a main means for diagnosing BV in clinical practice, with relatively high sensitivity. It diagnoses whether there is microecological imbalance by evaluating the microecological balance in the genital tract. However, due to over-reliance on the empirical judgment of experimental personnel, the Nugent scoring method has strong subjectivity, high requirements for the experience of inspectors, low inspection sensitivity and specificity, and a certain misdiagnosis or missed diagnosis rate. The use of test strips provides the possibility for on-site rapid detection. The operation of test strips is simple and does not require professional technical training, greatly saving time and labor costs. With the continuous increase in the demand for rapid microbial detection, the requirements for test strip preparation technology are also getting higher and higher. To enable the test strip to specifically identify microorganisms, it is necessary to precisely control the spraying uniformity of the detection reagent during preparation. In the existing test strip preparation process, it is difficult to timely adjust the subsequent preparation process according to the differences of nitrocellulose membranes, affecting the specificity and accuracy of detection. Therefore, improving the reliability of the test strip is a technical problem to be urgently solved.

[0003] For example, Chinese Patent Grant Publication No.: CN114280046B. This invention discloses a colorimetric-photothermal dual-mode test strip for detecting microorganisms and a preparation method thereof, including the following steps: preparing composite nanoparticles, adsorbing the antibody against the analyte, i.e., the primary antibody, on the surface to obtain a signal probe; assembling the test strip, fixing the antibody against the analyte and the secondary antibody on the detection area and the quality control area respectively; after mixing the signal probe with the analyte sample solution, inserting the sample pad end of the test strip into the mixed solution for chromatography; detecting the colorimetric mode: the color depth in the detection area is inversely proportional to the concentration of the analyte in the sample, and qualitative analysis is carried out by the naked eye and quantitative analysis is carried out by gray-scale analysis; photothermal mode: irradiating the detection area with a laser and collecting the temperature with a thermal imaging or temperature measurement device, and quantitative analysis is carried out according to the fact that the temperature in the detection area is directly proportional to the concentration of the analyte in the sample.

[0004] The following problems also exist in the prior art:

[0005] The prior art does not consider the nitrocellulose membrane as the core material of the test strip, and there will be certain differences in its water absorption. The curing rates of the nitrocellulose membranes with differences are different after being coated with glue, which will affect the fitting of the conjugate pad and the nitrocellulose membrane, thus affecting the spraying uniformity of the colored microspheres. In the process of preparing the test strip in the prior art, the risk tendency of abnormal fitting of the nitrocellulose membrane cannot be quickly identified, and the spraying method of the colored microspheres cannot be adaptively adjusted, which affects the spraying uniformity of the colored microspheres and the reliability of the preparation of the test strip. Summary of the Invention

[0006] Therefore, the present invention provides a test strip for quickly detecting microorganisms and a preparation method thereof, so as to overcome the problems in the prior art that in the process of preparing the test strip, the risk tendency of abnormal fitting of the nitrocellulose membrane cannot be quickly identified, the spraying method of the colored microspheres cannot be adaptively adjusted, which affects the spraying uniformity of the colored microspheres and the reliability of the preparation of the test strip.

[0007] To achieve the above object, the present invention provides a method for preparing a test strip for quickly detecting microorganisms, including:

[0008] Paste the nitrocellulose membrane required for the test strip on the plastic bottom plate, and paste the conjugate pad for fixing the colored microspheres to one end of the nitrocellulose membrane with a preset overlapping width;

[0009] Roll press the pasted conjugate pad, obtain a plurality of frame contour images of the overlapping area within a preset acquisition period, and determine the resilience coefficient according to the comparison of different frame contour images to determine whether there is a risk of abnormal fitting in the overlapping area;

[0010] In response to the existence of a risk of abnormal fitting, determine the contour characterization value according to the contour image of the characteristic frame within the preset acquisition period, and determine the fitting characterization parameter according to the comparison of a plurality of contour characterization values to determine the spraying abnormal category of the overlapping area;

[0011] Select the spraying adjustment method of the colored microspheres according to the spraying abnormal category, including,

[0012] Adjust the spraying angle of the colored microspheres sprayed onto the conjugate pad according to the resilience coefficient;

[0013] And, turn on the vibration device of the plastic bottom plate during the spraying process, and adjust the vibration frequency of the vibration device according to the fitting characterization parameter;

[0014] Paste the sample pad and the absorbent pad at the preset positions on the plastic bottom plate, and connect them to the nitrocellulose membrane and the conjugate pad to obtain the target test strip.

[0015] Furthermore, it is characterized in that determining the resilience coefficient includes,

[0016] Calculate the overlapping area value of adjacent frame contour images within a preset acquisition period, and determine the minimum value of the overlapping area value as the rebound coefficient.

[0017] Further, determining whether there is a risk of abnormal fitting in the overlapping area includes

[0018] If the rebound coefficient of the overlapping area meets the fitting risk determination condition, it is determined that there is a risk of abnormal fitting in the overlapping area;

[0019] If the rebound coefficient of the overlapping area does not meet the fitting risk determination condition, it is determined that there is no risk of abnormal fitting in the overlapping area;

[0020] Wherein, the fitting risk determination condition is that the rebound coefficient does not exceed a preset rebound coefficient reference value.

[0021] Further, the process of determining the fitting characterization parameter includes

[0022] Obtain a number of contour characterization values on the contour image of the feature frame, and determine the variance of the contour characterization values as the fitting characterization parameter;

[0023] Wherein, the feature frame is the last frame within a preset acquisition period, and the contour characterization value is the value in the width direction of the overlapping area.

[0024] Further, determining the spraying abnormality category of the overlapping area includes

[0025] If the fitting characterization parameter of the overlapping area meets the first abnormality determination condition, the spraying abnormality category of the overlapping area is determined as the first spraying abnormality category;

[0026] If the fitting characterization parameter of the overlapping area does not meet the first abnormality determination condition, the spraying abnormality category of the overlapping area is determined as the second spraying abnormality category;

[0027] Wherein, the first abnormality determination condition is that the fitting characterization parameter does not exceed a preset fitting characterization reference value.

[0028] Further, selecting the spraying adjustment method for the colored microspheres includes

[0029] If the spraying abnormality category is the first spraying abnormality category, select the spraying adjustment method for the colored microspheres as adjusting the spraying angle of the colored microspheres sprayed onto the bonding pad according to the rebound coefficient;

[0030] If the spraying abnormality category is the second spraying abnormality category, select the spraying adjustment method for the colored microspheres as turning on the vibration device for the plastic bottom plate during spraying and adjusting the vibration frequency of the vibration device according to the fitting characterization parameter.

[0031] Furthermore, the adjustment amount of the spraying angle is positively correlated with the rebound coefficient.

[0032] Furthermore, the vibration frequency is positively correlated with the fitting characterization parameter.

[0033] The present invention also provides a test strip for rapid detection of microorganisms, including,

[0034] A detection sample layer, which includes a sample pad for filtering a sample solution and a binding pad for fixing colored microspheres;

[0035] A detection result display layer, which is connected to the detection sample layer and includes a nitrocellulose membrane for displaying a detection result;

[0036] An absorbent layer, which is connected to the detection result display layer and includes an absorbent pad for absorbing excess sample solution.

[0037] Furthermore, a monoclonal antibody for STD items for detecting the microorganism to be detected and a monoclonal antibody against Lactobacillus are provided on the colored microspheres, wherein the monoclonal antibody for STD items is one of a monoclonal antibody of Gardner, a monoclonal antibody of Neisseria gonorrhoeae, a monoclonal antibody of Trichomonas, a monoclonal antibody of Chlamydia, a monoclonal antibody of Mycoplasma, and a monoclonal antibody of Candida albicans.

[0038] Compared with the prior art, the beneficial effects of the present invention are as follows. By pasting the nitrocellulose membrane required for the test strip on a plastic bottom plate, pasting the binding pad for fixing colored microspheres to one end of the nitrocellulose membrane with a preset overlapping width, rolling the pasted binding pad, obtaining contour images of a plurality of overlapping regions within a preset acquisition period, determining the rebound coefficient according to the comparison of contour images of different frames to determine whether there is a risk of abnormal fitting in the overlapping region, in response to the existence of the risk of abnormal fitting, determining a contour characterization value according to the contour image of a characteristic frame within the preset acquisition period, determining a fitting characterization parameter according to the comparison of a plurality of contour characterization values to determine the spraying abnormal category of the overlapping region, selecting a spraying adjustment method for colored microspheres according to the spraying abnormal category, and pasting the sample pad and the absorbent pad to preset positions on the plastic bottom plate and connecting them to the nitrocellulose membrane and the binding pad to prepare a target test strip. Furthermore, during the preparation process of the test strip, the risk tendency of abnormal fitting of the nitrocellulose membrane is quickly identified, the spraying method of colored microspheres is adaptively adjusted, the spraying uniformity of colored microspheres and the reliability of the preparation of the detection test strip are improved.

[0039] In particular, the present invention obtains contour images of overlapping areas of several frames within a preset acquisition cycle by rolling the pasted binding pad. It can be understood that the rolling process will cause the overlapping area of the binding pad and the nitrocellulose membrane to be subjected to a certain pressure. Obtaining the contour image can intuitively reflect the morphological changes of the overlapping area after rolling. By analyzing the contour images of different frames, the degree of fit between the binding pad and the nitrocellulose membrane can be determined. If the fit is good, the contour image will show a relatively stable and regular shape; if there is an abnormal fit, the contour image will show corresponding irregular changes, thereby timely discovering potential problems and ensuring the quality of the test paper. The present invention obtains contour images of overlapping areas of several frames within a preset acquisition cycle by rolling the pasted binding pad. Furthermore, it is achieved that the fitting state of the overlapping area can be quickly obtained during the preparation of the test paper, thereby improving the reliability of the preparation of the test paper.

[0040] In particular, the present invention determines the rebound coefficient based on the comparison of contour images of different frames to determine whether there is a risk of abnormal fitting in the overlapping area. It can be understood that by analyzing contour images of different frames to determine the rebound coefficient, the fitting state of the overlapping area can be converted into a specific numerical value, which makes the determination of the risk of abnormal fitting no longer rely on subjective observation and empirical judgment, but is based on objective data. If the rebound coefficient is within a preset range, it means that the water absorption of the nitrocellulose membrane is relatively uniform, and the overlapping area of the bonding pad and the nitrocellulose membrane is in good fitting state; if the rebound coefficient does not meet the range, it indicates that there may be a risk of loose fitting, which provides strong quantitative support for accurately evaluating the fitting quality and can detect the overlapping area at an early stage during the production process. Possible risks of abnormal fitting can be detected by continuously acquiring contour images and analyzing them within the preset acquisition cycle. Even slight changes or abnormalities in fitting can be detected, which helps to take timely measures for adjustment and improvement, and avoid discovering problems in subsequent production steps, thereby reducing material waste, improving production efficiency, and reducing production costs. By accurately determining the risk of abnormal fitting, it can be ensured that the quality of the fitting of the binding pad and the overlapping area of the nitrocellulose membrane of each batch of test strips can meet high standards, which helps to improve the accuracy of the test strip test results and enhance the quality stability and reliability of the product. Furthermore, in the preparation process of the test strip, it is possible to quickly identify the risk tendency of abnormal fitting of the nitrocellulose membrane and improve the reliability of the preparation of the test strips.

[0041] In particular, the present invention determines the contour characterization value based on the contour image of the feature frame within a preset acquisition period, and determines the spraying abnormality category of the overlapping area according to the comparison of several contour characterization values. It can be understood that by analyzing the values in the width direction of the overlapping area in the contour image of the feature frame as the contour characterization value, the key features of the overlapping area in the final state after rolling can be focused on. Calculating the variance of the contour characterization value as the fitting characterization parameter can quantify the change in the width direction of the overlapping area. If the fitting characterization parameter does not exceed the preset fitting characterization reference value, that is, the variance is small, it indicates that the numerical change in the width direction is small, and there is a problem of consistent abnormal fitting degree of the overall overlapping area. It is determined as the first spraying abnormality category. If the fitting characterization parameter exceeds the preset fitting characterization reference value, that is, the variance is large, it indicates that the numerical change in the width direction is large, and there is a problem of inconsistent abnormal fitting degree of the overall overlapping area. It is determined as the second spraying abnormality category, which helps to analyze different types of abnormalities targeted and provides an accurate basis for subsequent adjustment. The present invention determines the contour characterization value based on the contour image of the feature frame within a preset acquisition period, and determines the spraying abnormality category of the overlapping area according to the comparison of several contour characterization values. Furthermore, during the preparation process of the test strip, the risk tendency of abnormal fitting of the nitrocellulose membrane can be quickly identified, and the reliability of the preparation of the test strip is improved.

[0042] In particular, under the condition of the first spraying abnormality category, the present invention adjusts the spraying angle of the colored microspheres sprayed onto the conjugate pad according to the rebound coefficient. It can be understood that when the conjugate pad and the cellulose membrane are overlapped and pasted, there will be a slope with a certain height difference. Different curing rates of the glue in the overlapping area will generate an upward rebound force in the overlapping area, increasing the angle between the conjugate pad and the nitrocellulose membrane, thus causing the slope of the slope to become larger. Under the action of gravity, the microspheres are more likely to roll down along the slope and accumulate at the bottom of the slope, affecting the spraying uniformity of the colored microspheres. The first spraying abnormality category is that the angle of the slope increases uniformly as a whole. Adjusting the spraying angle will change the initial velocity direction and force condition when the colored microspheres are ejected. When the slope angle becomes larger, if spraying is still carried out at the original angle, the microspheres are more likely to quickly roll down along the slope to the bottom and accumulate under the influence of gravity and the slope angle. Furthermore, during the preparation process of the test strip, the risk tendency of abnormal fitting of the nitrocellulose membrane can be quickly identified, the spraying method of the colored microspheres can be adaptively adjusted, and the spraying uniformity of the colored microspheres and the reliability of the preparation of the test strip are improved.

[0043] In particular, under the condition of the second spraying abnormality category, during the spraying process, the vibration device for the plastic base plate is turned on, and the vibration frequency of the vibration device is adjusted according to the fitting characterization parameters. It can be understood that the condition of the second spraying abnormality category means that there are differences in the angle changes of the slope, and it does not increase uniformly as a whole. The slope with angle change differences will cause the colored microspheres to be more likely to locally aggregate, and the external force generated by vibration can give the colored microspheres additional kinetic energy. This kinetic energy enables the microspheres to overcome the frictional force on the slope surface and the adhesion force between each other, prompting them to roll and disperse from the accumulation area to the surrounding areas, and then roll to other areas of the slope, thereby reducing the accumulation amount at the bottom and making the distribution of the microspheres on the slope more uniform. Since the slopes at different positions are different, the distribution of the colored microspheres will also be uneven. The external vibration force can allow the microspheres to readjust their positions according to the actual shape and slope changes of the slope. During the vibration process, the microspheres will tend to move towards the areas with gentler slopes and larger spaces, thereby improving the overall distribution uniformity. In the area where the slope suddenly becomes steeper, the colored microspheres will move upward for a certain distance under the action of vibration to reach a more balanced distribution state. Furthermore, during the preparation process of the test strip, the risk tendency of the abnormal fitting of the nitrocellulose membrane can be quickly identified, the spraying method of the colored microspheres can be adaptively adjusted, the spraying uniformity of the colored microspheres can be improved, and the reliability of the preparation of the test strip can be enhanced. Description of the Drawings

[0044] Figure 1 It is a step diagram of the method for preparing a test strip for quickly detecting microorganisms according to an embodiment of the present invention;

[0045] Figure 2 It is a logic flowchart for determining whether there is a risk of abnormal fitting in the overlapping area according to an embodiment of the present invention;

[0046] Figure 3 It is a logic flowchart for determining the spraying abnormality category of the overlapping area according to an embodiment of the present invention;

[0047] Figure 4 It is a schematic diagram of a test strip for quickly detecting microorganisms according to an embodiment of the present invention;

[0048] In the figure: 1 - nitrocellulose membrane; 2 - conjugate pad; 3 - sample pad; 4 - absorbent pad. Detailed Embodiments

[0049] In order to make the objectives and advantages of the present invention clearer, the present invention will be further described below in conjunction with embodiments; it should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0050] The preferred embodiments of the present invention will be described below with reference to the accompanying drawings. Those skilled in the art should understand that these embodiments are only used to explain the technical principles of the present invention and do not limit the protection scope of the present invention.

[0051] It should be noted that in the description of the present invention, the terms indicating the direction or positional relationship such as "upper", "lower", "inner", "outer", etc. are based on the direction or positional relationship shown in the drawings. This is only for convenience of description and does not indicate or imply that the device or element must have a specific orientation, be constructed and operated in a specific orientation. Therefore, it should not be construed as a limitation to the present invention.

[0052] In addition, it should also be noted that in the description of the present invention, unless otherwise clearly specified and defined, the terms "installation" and "connection" should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be directly connected or indirectly connected through an intermediate medium, and it can be the communication inside two elements. For those skilled in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0053] Please refer to Figure 1 As shown, it is a step diagram of a method for preparing a test strip for rapid detection of microorganisms according to an embodiment of the present invention. A method for preparing a test strip for rapid detection of microorganisms includes:

[0054] Step S100: Paste the nitrocellulose membrane 1 required for the test strip on the plastic bottom plate, and paste the conjugate pad 2 for fixing the colored microspheres to one end of the nitrocellulose membrane 1 with a preset overlapping width.

[0055] Specifically, the preset overlapping width can be set by those skilled in the art according to the average value of several historical experimental data. The value range of the preset overlapping width can be [1, 5], and the interval unit is mm. If the value of the preset overlapping width is too large, it will cause too much overlap between the conjugate pad and the nitrocellulose membrane. On the one hand, it will increase the waste of materials and raise the production cost. On the other hand, it will make the flow path of the sample liquid on the test strip complex, affecting the migration speed and uniformity of the test sample solution on the test strip. If the value of the preset overlapping width is too small, the combination between the conjugate pad and the nitrocellulose membrane will not be firm enough. At the same time, it cannot ensure the smooth transfer of the colored microspheres from the conjugate pad to the nitrocellulose membrane, affecting the generation of detection signals. Preferably, the preset overlapping width can be 3 mm.

[0056] Step S200: Roll press the pasted conjugate pad 2, obtain the contour images of several overlapping regions within a preset acquisition period, and determine the resilience coefficient according to the comparison of different frame contour images to determine whether there is a risk of abnormal fitting in the overlapping region.

[0057] Specifically, the preset acquisition period can be set by those skilled in the art according to the accuracy requirements for the preparation of the test strip. The higher the accuracy requirement, the longer the preset acquisition period. Preferably, the preset acquisition period can be 2 min, and the moment when the rolling is completed is taken as the starting moment of the preset acquisition period.

[0058] Specifically, in the present invention, by rolling the bonded conjugate pad, contour images of a plurality of overlapping regions are obtained within the preset acquisition period. It can be understood that during the rolling process, a certain pressure is applied to the overlapping region of the conjugate pad and the nitrocellulose membrane. Obtaining the contour images can intuitively reflect the morphological changes of the overlapping region after rolling. By analyzing the contour images of different frames, the fitting degree between the conjugate pad and the nitrocellulose membrane can be judged. If the fitting is good, the contour images will present a relatively stable and regular form; if there is abnormal fitting, the contour images will show corresponding irregular changes, thereby timely discovering potential problems and ensuring the quality of the test strip. In the present invention, by rolling the bonded conjugate pad, contour images of a plurality of overlapping regions are obtained within the preset acquisition period. Furthermore, during the preparation process of the test strip, the fitting state of the overlapping region is quickly obtained, and the reliability of the preparation of the test strip is improved.

[0059] Step S300, in response to the risk of abnormal fitting, determine the contour characterization value according to the contour images of the characteristic frames within the preset acquisition period, and determine the fitting characterization parameter according to the comparison situation of a plurality of contour characterization values, so as to determine the spraying abnormal category of the overlapping region;

[0060] Step S400, select the spraying adjustment method of the colored microspheres according to the spraying abnormal category, including,

[0061] Adjust the spraying angle of the colored microspheres sprayed onto the conjugate pad according to the rebound coefficient;

[0062] And, during the spraying process, turn on the vibration device of the plastic bottom plate, and adjust the vibration frequency of the vibration device according to the fitting characterization parameter;

[0063] Step S500, paste the sample pad 3 and the absorbent pad 4 at the preset positions on the plastic bottom plate, and connect them to the nitrocellulose membrane 1 and the conjugate pad 2 to prepare the target test strip.

[0064] Specifically, IgY antibodies are also provided on the colored microspheres.

[0065] Specifically, a C-line quality control area containing goat anti-chicken polyclonal antibody, a detection display area containing the STD test item, and a detection display area containing lactobacillus can be provided on the nitrocellulose membrane.

[0066] Specifically, the essence of vaginal infection is microecological imbalance. The ultimate treatment goal of vaginal infection is to restore vaginal microecological balance. In the vaginal microecology of healthy women, more than 95% of the flora are lactobacilli, mainly Lactobacillus jensenii, Lactobacillus crispatus, Lactobacillus gasseri, and Lactobacillus iners. Clinical studies have shown that after lactobacilli are implanted into the vagina, they can adhere and colonize on vaginal epithelial cells, produce lactic acid by decomposing glycogen, maintain the acidic environment of the vagina, and at the same time produce substances such as H2O2, bacteriocins, and lysozyme to inhibit the growth of pathogenic bacteria. Lactobacilli refer to a general term for a class of non-spore-forming, Gram-positive bacteria whose main product of fermenting sugars is lactic acid. Most of them are essential and have important physiological functions in the human body. They are widely present in the human intestine. Lactobacilli are facultative anaerobes and prefer to grow under anaerobic conditions, but they will not die because of contact with oxygen. In the present invention, monoclonal antibodies for detecting STD items to be measured, monoclonal antibodies against lactobacilli, and IgY antibodies can be set on the color microspheres. Furthermore, the simultaneous detection of multiple targets of microorganisms related to vaginitis is realized, the microecological health is evaluated, the detection process is simple, and the detection efficiency and reliability are improved.

[0067] Specifically, the sample pad is connected to the conjugate pad and is pasted on the other side of one end where the conjugate pad is connected to the nitrocellulose membrane. The width of the overlapping area between the sample pad and the conjugate pad can be 3 mm. The absorbent pad is pasted on the other end of the nitrocellulose membrane away from the conjugate pad and the sample pad and is connected to the nitrocellulose membrane. The width of the overlapping area between the absorbent pad and the nitrocellulose membrane can be 3 mm. Such a layout enables the sample to flow from the sample pad through the nitrocellulose membrane towards the absorbent pad. During this process, substances such as color microspheres in the conjugate pad react with the sample, thereby realizing the detection of the target substance in the sample, which will not be elaborated here.

[0068] Specifically, determining the resilience coefficient includes

[0069] Calculating the overlapping area value of adjacent frame contour images within a preset acquisition period, and determining the minimum value of the overlapping area value as the resilience coefficient.

[0070] Specifically, image preprocessing such as grayscale conversion and noise reduction can be performed on the acquired contour images, edge algorithms can be used for contour extraction, and the contour images can be converted into binary images according to the pixel counting method to obtain the overlapping area value, which will not be elaborated here.

[0071] Please refer to Figure 2 As shown, it is a logic flowchart for determining whether there is a risk of abnormal fitting in the overlapping area in an embodiment of the present invention. Determining whether there is a risk of abnormal fitting in the overlapping area includes

[0072] If the resilience coefficient of the overlapping area meets the fitting risk determination condition, it is determined that there is a risk of abnormal fitting in the overlapping area;

[0073] If the springback coefficient of the overlapping area does not meet the fitting risk determination condition, it is determined that there is no fitting abnormality risk in the overlapping area;

[0074] Among them, the fitting risk determination condition is that the springback coefficient does not exceed a preset springback coefficient reference value.

[0075] Specifically, the preset springback coefficient reference value is the product of the area before rolling of the overlapping area and the springback factor. The springback factor can be set by those skilled in the art according to the preparation accuracy requirements of the test strip. The higher the accuracy requirement, the smaller the springback factor. The value range of the springback factor can be [0.7, 0.9]. Preferably, the springback factor can be 0.8.

[0076] Specifically, the present invention determines the springback coefficient according to the comparison of different frame contour images to determine whether there is a fitting abnormality risk in the overlapping area. It can be understood that by analyzing different frame contour images to determine the springback coefficient, the fitting state of the overlapping area can be converted into specific numerical values, which makes the determination of the fitting abnormality risk no longer rely on subjective observation and empirical judgment, but on objective data. If the springback coefficient is within the preset range, it indicates that the water absorption of the nitrocellulose membrane is relatively uniform, and the fitting state of the conjugate pad and the nitrocellulose membrane overlapping area is good; if the springback coefficient does not meet this range, it indicates that there may be a risk of loose fitting, providing strong quantitative support for accurately evaluating the fitting quality. During the production process, the possible fitting abnormality risk in the overlapping area can be detected at an early stage. Since the contour images are continuously acquired and analyzed within the preset acquisition period, even minor fitting changes or abnormalities can be detected, which helps to take timely measures for adjustment and improvement, avoiding problems being discovered only in subsequent production steps, thereby reducing material waste, improving production efficiency, and reducing production costs. By accurately determining the fitting abnormality risk, it can ensure that the fitting quality of the overlapping area between the conjugate pad and the nitrocellulose membrane of each batch of test strips can reach a high standard, helping to improve the accuracy of the test strip detection results, enhancing the quality stability and reliability of the product. Furthermore, during the preparation process of the test strip, the fitting abnormality risk tendency of the nitrocellulose membrane can be quickly identified, improving the reliability of the preparation of the test strip.

[0077] Specifically, the nitrocellulose membrane is a porous material with certain non-uniformities in the pore size, pore distribution, and fiber arrangement inside. These differences in microstructure result in different water absorption capacities in different parts. When glue is applied, the curing process of the glue is usually affected by water. Areas with strong water absorption will absorb more water, delaying the curing reaction of the glue and resulting in a slower curing rate. In areas with weak water absorption, the glue is less affected by water and the curing rate is relatively fast. When rolling the bonded pad after pasting, the bonded pad and the nitrocellulose membrane will be deformed under the action of external force. Due to the non-uniform water absorption of the nitrocellulose membrane and the different curing rates of the glue after application, after the external rolling force disappears, the material will have different degrees of rebound due to different curing degrees. In the part with a fast curing rate, the bonding structure formed by the glue is relatively stable and the rebound amount is small. In the part with a slow curing rate, the bonding structure is not yet completely stable and there will be a large rebound. The overlapping area value of adjacent frame contour images can intuitively reflect the shape change of the overlapping area. The larger the overlapping area, the smaller the shape change of the overlapping area at adjacent moments, that is, the smaller the rebound amount. On the contrary, the smaller the overlapping area, the greater the shape change and the greater the rebound amount. Determining the minimum value of the overlapping area value as the rebound coefficient can reflect the situation where the shape change of the overlapping area is the largest during the entire rebound process. Furthermore, during the preparation process of the test strip, the risk tendency of abnormal fitting of the nitrocellulose membrane can be quickly identified, improving the reliability of the preparation of the test strip.

[0078] Specifically, the process of determining the fitting characterization parameter includes

[0079] Obtaining a number of contour characterization values on the contour image of the feature frame, and determining the variance of the contour characterization values as the fitting characterization parameter.

[0080] Among them, the feature frame is the last frame within a preset acquisition period, and the contour characterization value is the value in the width direction of the overlapping area.

[0081] Please refer to Figure 3 As shown, it is the logic flow chart for determining the spraying abnormal category of the overlapping area in the embodiment of the present invention. Determining the spraying abnormal category of the overlapping area includes

[0082] If the fitting characterization parameter of the overlapping area meets the first abnormal determination condition, then determine the spraying abnormal category of the overlapping area as the first spraying abnormal category;

[0083] If the fitting characterization parameter of the overlapping area does not meet the first abnormal determination condition, then determine the spraying abnormal category of the overlapping area as the second spraying abnormal category;

[0084] Among them, the first abnormal determination condition is that the fitting characterization parameter does not exceed the preset fitting characterization reference value.

[0085] Specifically, the preset fitting characterization reference value can be set by those skilled in the art according to the precision requirements for the preparation of the test strip. The higher the precision requirements, the smaller the preset fitting characterization reference value. Preferably, the fitting characterization reference value can be 0.5.

[0086] Specifically, the present invention determines the contour characterization value based on the contour image of the feature frame within the preset acquisition period, and determines the spraying abnormality category of the overlapping area according to the comparison of several contour characterization values. It can be understood that by analyzing the numerical values in the width direction of the overlapping area in the contour image of the feature frame as the contour characterization value, the key features of the overlapping area in the final state after rolling can be focused on. Calculating the variance of the contour characterization value as the fitting characterization parameter can quantify the change in the width direction of the overlapping area. If the fitting characterization parameter does not exceed the preset fitting characterization reference value, that is, the variance is small, it indicates that the numerical change in the width direction is small, and there is a problem of consistent abnormal fitting degree of the overall overlapping area, and it is determined as the first spraying abnormality category. If the fitting characterization parameter exceeds the preset fitting characterization reference value, that is, the variance is large, it indicates that the numerical change in the width direction is large, and there is a problem of inconsistent abnormal fitting degree of the overall overlapping area, and it is determined as the second spraying abnormality category, which helps to analyze different types of abnormalities targeted and provide an accurate basis for subsequent adjustment. The present invention determines the contour characterization value based on the contour image of the feature frame within the preset acquisition period, and determines the spraying abnormality category of the overlapping area according to the comparison of several contour characterization values. Furthermore, during the preparation process of the test strip, the risk tendency of abnormal fitting of the nitrocellulose membrane can be quickly identified, and the reliability of the preparation of the test strip can be improved.

[0087] Specifically, the selected spraying adjustment method for the colored microspheres includes

[0088] If the spraying abnormality category is the first spraying abnormality category, the selected spraying adjustment method for the colored microspheres is to adjust the spraying angle of the colored microspheres sprayed onto the conjugate pad according to the rebound coefficient;

[0089] If the spraying abnormality category is the second spraying abnormality category, the selected spraying adjustment method for the colored microspheres is to turn on the vibration device for the plastic bottom plate during the spraying process and adjust the vibration frequency of the vibration device according to the fitting characterization parameter.

[0090] Specifically, the vibration device can be an electromagnetic vibration device, which works based on the principle of electromagnetic induction. An alternating magnetic field is generated by an alternating current, and the vibrating component with magnetism makes a reciprocating motion under the action of the magnetic field force to generate vibration, which will not be elaborated here.

[0091] Specifically, the adjustment amount of the spraying angle is positively correlated with the rebound coefficient.

[0092] Specifically, a nozzle that adjusts the spraying angle by motor drive can be used. The microprocessor is utilized to control the motor to adjust the spraying angle. The direction perpendicular to the bonding pad of the nozzle is marked as 0°, and the initial spraying angle when the nozzle rotates 20° towards the nitrocellulose membrane direction is set. The value range of the spraying angle can be [20, 60], and the interval unit is °.

[0093] Specifically, under the condition of the first spraying abnormal category, the spraying angle of the colored microspheres sprayed onto the bonding pad is adjusted according to the rebound coefficient. It can be understood that when the bonding pad and the nitrocellulose membrane are overlapped and pasted, there will be a slope with a certain height difference. Different curing rates of the glue in the overlapping area will generate an upward rebound force in the overlapping area, increasing the angle between the bonding pad and the nitrocellulose membrane, thereby causing the slope gradient to become larger. Under the action of gravity, the microspheres are more likely to roll down along the slope and accumulate at the bottom of the slope, affecting the spraying uniformity of the colored microspheres. The first spraying abnormal category is that the angle of the slope increases uniformly as a whole. Adjusting the spraying angle will change the initial velocity direction and the force condition when the colored microspheres are ejected. When the slope angle becomes larger, if spraying is still carried out at the original angle, the microspheres are more likely to roll down quickly along the slope to the bottom and accumulate under the influence of gravity and the slope angle. Furthermore, during the preparation process of the test strip, the risk tendency of abnormal bonding of the nitrocellulose membrane can be quickly identified, the spraying method of the colored microspheres can be adaptively adjusted, the spraying uniformity of the colored microspheres can be improved, and the reliability of the preparation of the test strip can be enhanced.

[0094] Specifically, the vibration frequency has a positive correlation with the bonding characterization parameter.

[0095] Specifically, the setting range of the vibration frequency can be [50, 150], and the interval unit is Hz.

[0096] Specifically, under the condition of the second spraying abnormality category, during the spraying process, the vibration device for the plastic base plate is turned on, and the vibration frequency of the vibration device is adjusted according to the fitting characterization parameters. It can be understood that the condition of the second spraying abnormality category means that there are differences in the angle changes of the slope, and it does not increase uniformly as a whole. The slope with angle change differences will cause the colored microspheres to be more likely to locally aggregate, and the external force generated by vibration can give the colored microspheres additional kinetic energy. This kinetic energy enables the microspheres to overcome the frictional force on the slope surface and the adhesion force between each other, prompting them to roll and disperse from the accumulation area to the surrounding areas, and then roll to other areas of the slope, thereby reducing the accumulation amount at the bottom and making the distribution of the microspheres on the slope more uniform. Since the slopes at different positions are different, the distribution of the colored microspheres will also be uneven. The external vibration force can allow the microspheres to readjust their positions according to the actual shape and slope changes of the slope. During the vibration process, the microspheres will tend to move to the areas with gentler slopes and larger spaces, thereby improving the overall distribution uniformity. In the area where the slope suddenly becomes steeper, the colored microspheres will move upward for a certain distance under the action of vibration to reach a more balanced distribution state. Furthermore, during the preparation process of the test strip, the risk tendency of abnormal fitting of the nitrocellulose membrane can be quickly identified, the spraying method of the colored microspheres can be adaptively adjusted, the spraying uniformity of the colored microspheres can be improved, and the reliability of the preparation of the test strip can be enhanced.

[0097] Please refer to Figure 4 as shown in the figure, which is a schematic diagram of the test strip for rapid detection of microorganisms according to an embodiment of the present invention. A test strip for rapid detection of microorganisms according to the present invention includes:

[0098] A detection sample layer, which includes a sample pad 3 for filtering the sample solution and a binding pad 2 for fixing colored microspheres;

[0099] A detection result display layer, which is connected to the detection sample layer and includes a nitrocellulose membrane 1 for displaying the detection result;

[0100] An absorbent layer, which is connected to the detection result display layer and includes an absorbent pad 4 for absorbing the excess sample solution.

[0101] Specifically, monoclonal antibodies for STD items for detecting the microorganisms to be detected and monoclonal antibodies against Lactobacillus are provided on the colored microspheres. Among them, the monoclonal antibody for the STD item is one of the monoclonal antibodies of Gardner, gonococcus, trichomonas, chlamydia, mycoplasma, and Candida albicans.

[0102] Specifically, by using semi-quantitative detection of Lactobacillus S-layer protein or N-protein to assist in the diagnosis of microecological detection, the microecological health can be characterized. In the sample to be tested, if Lactobacillus is strongly positive and the content of the pathogen antigen to be tested is small, it indicates that the sample population is in a state of Lactobacillus dominant bacteria and has no BV indication; in the sample to be tested, if Lactobacillus is weakly positive and the content of the pathogen antigen to be tested is obvious, it indicates that the sample population is in a state of pathogenic bacteria dominant bacteria and has a BV indication; in the sample to be tested, if Lactobacillus is negative and the content of the pathogen antigen to be tested is obvious, it indicates that the sample population is in a state of pathogenic bacteria dominant bacteria and has a severe BV indication.

[0103] Exemplarily, a specific embodiment of using Lactobacillus for vaginitis detection is given here. When the proportion of Lactobacillus is greater than 80% and Gardnerella is less than 5%, the population is healthy and has no bacterial vaginitis; when the proportion of Lactobacillus is less than 80% and greater than 60%, and Gardnerella is less than 20%, the population is in a transitional state and is very likely to further develop into bacterial vaginitis; when the proportion of Lactobacillus is less than 60% or Gardnerella is greater than 20%, the population is a patient with bacterial vaginitis.

[0104] So far, the technical solution of the present invention has been described in conjunction with the preferred embodiments shown in the drawings. However, it is easy for those skilled in the art to understand that the protection scope of the present invention is obviously not limited to these specific embodiments. Without departing from the principle of the present invention, those skilled in the art can make equivalent changes or substitutions to the relevant technical features, and the technical solutions after these changes or substitutions will all fall within the protection scope of the present invention.

[0105] The above are only the preferred embodiments of the present invention and are not used to limit the present invention; for those skilled in the art, the present invention can have various changes and modifications. Any modification, equivalent substitution, 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 preparing a test strip for rapid detection of microorganisms, characterized in that, Including: Pasting the nitrocellulose membrane required for the test strip on the plastic base plate, and pasting the binding pad for fixing the colored microspheres to one end of the nitrocellulose membrane with a preset overlapping width; Rolling the pasted binding pad, obtaining a plurality of contour images of the overlapping area within a preset acquisition period, and determining the resilience coefficient according to the comparison of different frame contour images to determine whether there is a risk of abnormal fitting in the overlapping area; Determining the resilience coefficient includes calculating the overlapping area value of adjacent frame contour images within a preset acquisition period, and determining the minimum value of the overlapping area value as the resilience coefficient; In response to the existence of a risk of abnormal fitting, determining the contour characterization value according to the contour image of the characteristic frame within a preset acquisition period, and determining the fitting characterization parameter according to the comparison of a plurality of contour characterization values to determine the spraying abnormal category of the overlapping area; The process of determining the fitting characterization parameter includes obtaining a plurality of contour characterization values on the contour image of the characteristic frame, and determining the variance of the contour characterization values as the fitting characterization parameter. The characteristic frame is the last frame within a preset acquisition period, and the contour characterization value is the value in the width direction of the overlapping area; If the fitting characterization parameter of the overlapping area meets the first abnormal determination condition, then determine the spraying abnormal category of the overlapping area as the first spraying abnormal category; If the fitting characterization parameter of the overlapping area does not meet the first abnormal determination condition, then determine the spraying abnormal category of the overlapping area as the second spraying abnormal category; The first abnormal determination condition is that the fitting characterization parameter does not exceed the preset fitting characterization reference value; Selecting the spraying adjustment method of the colored microspheres according to the spraying abnormal category, including, If the spraying abnormal category is the first spraying abnormal category, then adjust the spraying angle of the colored microspheres sprayed onto the binding pad according to the resilience coefficient; If the spraying abnormal category is the second spraying abnormal category, then turn on the vibration device of the plastic base plate during spraying, and adjust the vibration frequency of the vibration device according to the fitting characterization parameter; Pasting the sample pad and the absorbent pad at the preset positions on the plastic base plate, connecting them to the nitrocellulose membrane and the binding pad to obtain the target test strip.

2. The method for preparing a test strip for rapid detection of microorganisms according to claim 1, characterized in that, Determining whether there is a risk of abnormal fitting in the overlapping area includes, If the resilience coefficient of the overlapping area meets the fitting risk determination condition, then determine that the overlapping area has a risk of abnormal fitting; If the resilience coefficient of the overlapping area does not meet the fitting risk determination condition, then determine that the overlapping area does not have a risk of abnormal fitting; Wherein, the fitting risk determination condition is that the resilience coefficient does not exceed the preset resilience coefficient reference value.

3. The method for preparing a test strip for rapid detection of microorganisms according to claim 2, characterized in that, The adjustment amount of the spraying angle is positively correlated with the resilience coefficient.

4. The method for preparing a test strip for rapid detection of microorganisms according to claim 3, characterized in that, The vibration frequency is positively correlated with the fitting characterization parameter.

5. A test strip for rapid detection of microorganisms, which is prepared by using the method for preparing a test strip for rapid detection of microorganisms according to any one of claims 1-4 above, and is characterized in that, Including: A detection sample layer, which includes a sample pad for filtering the sample solution and a binding pad for fixing the colored microspheres; A detection result display layer, which is connected to the detection sample layer and includes a nitrocellulose membrane for displaying the detection result; An absorbent layer, which is connected to the detection result display layer and includes an absorbent pad for absorbing the excess sample solution.

6. The test strip for rapid detection of microorganisms according to claim 5, characterized in that, The color microspheres are provided with monoclonal antibodies for detecting STD items of the to-be-detected microorganisms and monoclonal antibodies against Lactobacillus, wherein the monoclonal antibody for the STD item is one of the monoclonal antibodies of Gardnerella, Neisseria gonorrhoeae, Trichomonas, Chlamydia trachomatis, Mycoplasma, and Candida albicans.

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