Test paper for rapidly detecting microorganisms and preparation method thereof

Abstract

The invention relates to the technical field of microorganism test paper preparation, in particular to test paper for rapidly detecting microorganisms and a preparation method thereof.The method includes the steps that a combination pad used for fixing color microspheres is pasted to one end of a nitrocellulose membrane, the pasted combination pad is rolled, and a plurality of frames of contour images of overlapped areas are obtained; determining a rebound coefficient according to the comparison condition of different frame contour images so as to judge whether there is an abnormal fitting risk, determining fitting characterization parameters according to the contour image of the feature frame in the preset acquisition period so as to determine the spraying abnormality category of the overlapping region, selecting a spraying adjustment mode of the color microspheres according to the spraying abnormality category, and adjusting the color microspheres according to the spraying adjustment mode. And the sample pad and the water absorption pad are connected with the nitrocellulose membrane and the combination pad to prepare the target test paper, so that the rapid identification of the abnormal fitting risk tendency of the nitrocellulose membrane and the adaptive adjustment of the spraying mode of the color microspheres are realized, and the preparation reliability of the detection test paper is improved.

Description

Technical Field

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

[0002] Reproductive tract infection has always been a concern for reproductive health at home and abroad. It is an inflammation caused by changes in the biochemical properties of microbial environmental flora and secretions. The essence of reproductive tract infection is microecological imbalance. At present, the Nugent scoring method is a major means of diagnosing BV in clinical practice. It has high sensitivity and diagnoses whether there is microecological imbalance by evaluating the microecological balance in the reproductive tract. However, the Nugent scoring method is overly dependent on the experience and judgment of the experimenters, which is highly subjective and requires high experience of the inspectors. The sensitivity and specificity of the examination are low, and there is a certain misdiagnosis or missed diagnosis rate. The use of test strips provides the possibility of rapid on-site detection. The test strips are simple to operate and do not require professional technical training, which greatly saves time and labor costs. With the increasing demand for rapid microbial detection, the requirements for the preparation technology of test strips are also getting higher and higher. In order to enable the test strips to specifically identify microorganisms, the spraying uniformity of the detection reagent must be accurately controlled during preparation. In the existing test strip preparation process, it is difficult to adjust the subsequent preparation process in time according to the different nitrocellulose membranes, which affects the specificity and accuracy of the detection. Therefore, improving the reliability of the test strips is a technical problem that needs to be solved urgently.

[0003] For example, the Chinese patent authorization announcement number is: CN114280046B, which discloses a colorimetric-photothermal dual-mode test strip for detecting microorganisms and a preparation method thereof, comprising the following steps: preparing composite nanoparticles, adsorbing the antibody to be detected, i.e., the primary antibody, on the surface to obtain a signal probe; assembling the test strip, fixing the antibody to be detected and the secondary antibody on the detection area and the quality control area, respectively; after mixing the signal probe with the sample liquid to be detected, inserting the sample pad end of the test strip into the mixed liquid for chromatography; detecting colorimetric mode: the color depth of the detection area is inversely proportional to the concentration of the sample to be detected, which can be qualitatively determined by the naked eye and quantitatively determined by grayscale analysis; photothermal mode: irradiating the detection area with a laser and collecting the temperature with a thermal imaging or temperature measuring device, and performing quantitative analysis based on the fact that the temperature of the detection area is proportional to the concentration of the sample to be detected.

[0004] The prior art still has the following problems: The prior art does not consider nitrocellulose membrane as the core material of the test paper, and its water absorption will have certain differences. The curing rate of the different nitrocellulose membrane after coating with glue is different, which will affect the bonding between the binding pad and the nitrocellulose membrane, thereby affecting the spraying uniformity of the colored microspheres. In the preparation process of the test paper, the prior art cannot quickly identify the abnormal bonding risk tendency of the nitrocellulose membrane, and cannot adaptively adjust the spraying method of the colored microspheres, which affects the spraying uniformity of the colored microspheres and the reliability of the preparation of the test paper. Summary of the invention

[0005] To this end, the present invention provides a test paper for rapid detection of microorganisms and a preparation method thereof, so as to overcome the problems in the prior art that, during the preparation process of the test paper, 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, and the spraying uniformity of the colored microspheres and the reliability of the preparation of the test paper are affected.

[0006] To achieve the above object, the present invention provides a method for preparing a test paper for rapid detection of microorganisms, comprising: Paste the nitrocellulose membrane required for the test paper on the plastic base plate, and paste the binding pad for fixing the colored microspheres to one end of the nitrocellulose membrane with a preset overlapping width; The pasted bonding pad is rolled, and contour images of overlapping areas of several frames are obtained within a preset acquisition period. The rebound coefficient is determined according to the comparison of contour images of different frames to determine whether there is a risk of abnormal bonding in the overlapping area; In response to the presence of a risk of abnormal lamination, a contour characterization value is determined according to a contour image of a feature frame within a preset acquisition period, and a lamination characterization parameter is determined according to a comparison of a plurality of contour characterization values ​​to determine a spraying abnormality category of the overlapping area; The spraying adjustment method of the colored microspheres is selected according to the spraying abnormality category, including: Adjusting the spraying angle of the colored microspheres onto the bonding pad according to the rebound coefficient; and, during the spraying process, turning on a vibration device for the plastic base plate, and adjusting the vibration frequency of the vibration device according to the bonding characterization parameter; The sample pad and the absorbent pad are pasted to the preset position of the plastic bottom plate, and connected to the nitrocellulose membrane and the binding pad to obtain the target test paper.

[0007] Further, it is characterized in that determining the coefficient of resilience comprises, The overlap area values ​​of the contour images of adjacent frames within a preset acquisition period are calculated, and the minimum value of the overlap area values ​​is determined as the rebound coefficient.

[0008] Further, determining whether there is a risk of abnormal bonding in the overlapping area includes: If the coefficient of resilience 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; If the coefficient of resilience 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; The bonding risk determination condition is that the rebound coefficient does not exceed a preset rebound coefficient reference value.

[0009] Further, the process of determining the fit characterization parameters includes, Acquire a plurality of contour characterization values ​​on the contour image of the feature frame, and determine the variance of the contour characterization values ​​as a fitting characterization parameter; The feature frame is the last frame in a preset acquisition period, and the contour characterization value is a value in the width direction of the overlapping area.

[0010] Further, the spraying anomaly categories of the overlapped area are determined to include: If the lamination 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; If the lamination 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; The first abnormality determination condition is that the fitting characterization parameter does not exceed a preset fitting characterization reference value.

[0011] Further, the spraying adjustment method of the selected colored microspheres includes: If the spraying abnormality category is the first spraying abnormality category, the spraying adjustment method of the selected colored microspheres is to adjust the spraying angle of the colored microspheres to the bonding pad according to the rebound coefficient; If the spraying abnormality category is the second spraying abnormality category, the spraying adjustment method of the selected colored microspheres is to turn on the vibration device of the plastic base plate during the spraying process, and adjust the vibration frequency of the vibration device according to the fitting characterization parameter.

[0012] Furthermore, the adjustment amount of the spraying angle is positively correlated with the rebound coefficient.

[0013] Furthermore, the vibration frequency is positively correlated with the fitting characterization parameter.

[0014] The present invention also provides a test paper for rapid detection of microorganisms, comprising: A detection sample layer, which includes a sample pad for filtering the sample solution and a binding pad for fixing the colored microspheres; A test result display layer, which is connected to the test sample layer and includes a nitrocellulose membrane for displaying the test result; The water absorption layer is connected to the detection result display layer and includes a water absorption pad for absorbing excess sample solution.

[0015] Furthermore, the colored microspheres are provided with monoclonal antibodies for detecting STD items of the microorganisms to be tested, and monoclonal antibodies against lactobacilli, wherein the monoclonal antibodies for STD items are one of the monoclonal antibodies of Gardnerella, monoclonal antibodies of gonococci, monoclonal antibodies of Trichomonas, monoclonal antibodies of Chlamydia, monoclonal antibodies of Mycoplasma, and monoclonal antibodies of Candida albicans.

[0016] Compared with the prior art, the beneficial effect of the present invention lies in that the present invention adheres the nitrocellulose membrane required for the test paper to a plastic base plate, adheres the binding pad for fixing the colored microspheres to one end of the nitrocellulose membrane with a preset overlapping width, rolls the pasted binding pad, obtains contour images of overlapping areas of several frames within a preset acquisition period, determines 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 area, determines a contour characterization value according to the contour image of the feature frame within the preset acquisition period in response to the existence of the abnormal fitting risk, determines the fitting characterization parameter according to the comparison of several contour characterization values ​​to determine the spraying abnormality category of the overlapping area, selects the spraying adjustment method of the colored microspheres according to the spraying abnormality category, adheres the sample pad and the absorbent pad to the preset position of the plastic base plate, and connects them to the nitrocellulose membrane and the binding pad to obtain the target test paper, thereby realizing the rapid identification of the abnormal fitting risk tendency of the nitrocellulose membrane in the preparation process of the test paper, adaptively adjusting the spraying method of the colored microspheres, improving the spraying uniformity of the colored microspheres and the reliability of the preparation of the test paper.

[0017] 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.

[0018] 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.

[0019] 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 based on the comparison of several contour characterization values. It can be understood that by analyzing the numerical value in the width direction of the overlapping area in the contour image of the feature frame as the contour characterization value, it is possible to focus on the key features of the overlapping area in the final state after rolling, and calculate the variance of the contour characterization value as the fitting characterization parameter, so as to 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 means that the numerical value change in the width direction is small, and there is a problem of consistent overall fitting abnormality in the overlapping area, and it is confirmed. 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 means that the value changes greatly in the width direction, and there is a problem of inconsistent overall fitting abnormality in the overlapping area. It is determined as the second spraying abnormality category, which is helpful for targeted analysis of different types of abnormalities and provides an accurate basis for subsequent adjustments. The present invention determines the contour characterization value through 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, in the preparation process of the test paper, it is realized that the risk tendency of fitting abnormality of the nitrocellulose membrane can be quickly identified, and the reliability of the preparation of the test paper is improved.

[0020] In particular, under the condition of the first spraying abnormality category, the present invention adjusts the spraying angle of the colored microspheres to the binding pad according to the rebound coefficient. It can be understood that when the binding pad and the cellulose membrane are overlapped and pasted, there will be a slope with a certain height difference. The different glue curing rates in the overlapping area will generate an upward rebound force in the overlapping area, which increases the angle between the binding pad and the nitrocellulose membrane, thereby increasing the slope gradient. The microspheres are more likely to roll down the slope under the action of gravity and accumulate at the bottom of the slope, affecting the spraying uniformity of the colored microspheres. The first spraying abnormality category, that is, the angle of the slope is uniformly increased as a whole. Adjusting the spraying angle will change the initial velocity direction and force conditions of the colored microspheres when they are sprayed out. When the slope angle increases, if it is still sprayed at the original angle, the microspheres are affected by gravity and the slope angle, and are more likely to roll down the slope quickly to the bottom and accumulate. Therefore, in the preparation process of the test paper, the risk tendency of abnormal bonding of the nitrocellulose membrane is quickly identified, the spraying method of the colored microspheres is adaptively adjusted, and the spraying uniformity of the colored microspheres and the reliability of the preparation of the test paper are improved.

[0021] In particular, under the second spraying abnormality category condition, the present invention turns on the vibration device for the plastic base plate during the spraying process, and adjusts the vibration frequency of the vibration device according to the fitting characterization parameter. It can be understood that the second spraying abnormality category condition, that is, the angle change of the slope is different, and it is not uniformly increased as a whole. The slope with angle change difference will make the colored microspheres more likely to gather locally, and the external force generated by the vibration can give the colored microspheres additional kinetic energy. This kinetic energy enables the microspheres to overcome the friction force on the slope surface and the adhesion between each other, prompting them to roll and disperse from the accumulation area to the surrounding area, and then roll to other areas of the slope, thereby reducing the accumulation amount at the bottom, making the microspheres on the slope The distribution is more uniform. The slopes at different locations are different, and the distribution of colored microspheres will also be uneven. The vibration external force can make the microspheres readjust their positions according to the actual shape and slope changes of the slope. During the vibration process, the microspheres tend to move to areas with gentler slopes and larger spaces, thereby improving the overall distribution uniformity. In areas where the slope suddenly increases, the colored microspheres will move upward for a distance under the action of vibration to achieve a more balanced distribution state. Furthermore, in the preparation process of the test paper, the abnormal risk tendency 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 paper can be improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 A step diagram of a method for preparing a test paper for rapid detection of microorganisms according to an embodiment of the present invention; Figure 2 A logic flow chart for determining whether there is a risk of abnormal fitting in an overlapping area according to an embodiment of the present invention; Figure 3A logic flow chart for determining the spraying anomaly category of the overlapping area according to an embodiment of the present invention; Figure 4 A schematic diagram of a test paper for rapid detection of microorganisms according to an embodiment of the present invention; In the figure: 1- nitrocellulose membrane; 2- binding pad; 3- sample pad; 4- absorbent pad. DETAILED DESCRIPTION

[0023] In order to make the objects and advantages of the present invention more clearly understood, the present invention is 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.

[0024] The preferred embodiments of the present invention are described below with reference to the accompanying drawings. It should be understood by those skilled in the art that these embodiments are only used to explain the technical principles of the present invention and are not intended to limit the protection scope of the present invention.

[0025] It should be noted that, in the description of the present invention, terms such as "upper", "lower", "inside" and "outside" indicating directions or positional relationships are based on the directions or positional relationships shown in the drawings. This is only for the 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 cannot be understood as a limitation on the present invention.

[0026] In addition, it should be noted that in the description of the present invention, unless otherwise clearly specified and limited, 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 a direct connection, or it can be an indirect connection through an intermediate medium, or it can be the internal communication of two components. For those skilled in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0027] See also Figure 1 As shown, it is a step diagram of a method for preparing a test paper for rapid detection of microorganisms according to an embodiment of the present invention. A method for preparing a test paper for rapid detection of microorganisms comprises: Step S100, pasting the nitrocellulose membrane 1 required for the test paper on the plastic base plate, and pasting the binding pad 2 for fixing the colored microspheres to one end of the nitrocellulose membrane 1 with a preset overlapping width; Specifically, the preset overlap width can be set by a technician in this field based on the average value of several historical experimental data. The value range of the preset overlap width can be [1, 5], and the unit of the interval is mm. If the value of the preset overlap width is too large, it will cause too much overlap between the binding pad and the nitrocellulose membrane, which will increase material waste and increase production costs on the one hand, and on the other hand, it will make the flow path of the sample liquid on the test paper complicated, affecting the migration speed and uniformity of the sample solution on the test paper. If the value of the preset overlap width is too small, the binding pad and the nitrocellulose membrane will not be firmly bonded. At the same time, it cannot ensure that the colored microspheres are smoothly transferred from the binding pad to the nitrocellulose membrane, affecting the generation of the detection signal. Preferably, the preset overlap width can be 3mm.

[0028] Step S200, rolling the pasted bonding pad 2, acquiring contour images of overlapping regions of several frames within a preset acquisition period, and determining the rebound coefficient according to the comparison of contour images of different frames to determine whether there is a risk of abnormal pasting in the overlapping region; Specifically, the preset collection period can be set by technical personnel in this field according to the accuracy requirements of the test paper preparation. The higher the accuracy requirements, the longer the preset collection period. Preferably, the preset collection period can be 2 minutes, with the moment when rolling is completed as the starting moment of the preset collection period.

[0029] Specifically, 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 present 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.

[0030] Step S300, in response to the presence of a risk of abnormal lamination, determining a contour characterization value according to a contour image of a feature frame within a preset acquisition period, and determining a lamination characterization parameter according to a comparison of a plurality of contour characterization values, so as to determine a spraying abnormality category of the overlapping area; Step S400, selecting a spraying adjustment method for colored microspheres according to the spraying abnormality category, including: Adjusting the spraying angle of the colored microspheres onto the bonding pad according to the rebound coefficient; and, during the spraying process, turning on a vibration device for the plastic base plate, and adjusting the vibration frequency of the vibration device according to the bonding characterization parameter; Step S500, sticking the sample pad 3 and the absorbent pad 4 to the preset position of the plastic bottom plate, connecting with the nitrocellulose membrane 1 and the binding pad 2, so as to obtain the target test paper.

[0031] Specifically, IgY antibodies are also disposed on the colored microspheres.

[0032] Specifically, a C-line quality control area containing goat anti-chicken polyclonal antibodies, a detection display area containing STD test items, and a detection display area containing lactobacilli can be set on the nitrocellulose membrane.

[0033] Specifically, the essence of vaginal infection is the imbalance of microecology. The ultimate treatment goal of vaginal infection is to restore the balance of vaginal microecology. More than 95% of the flora in the vaginal microecology of healthy women are lactobacilli, mainly Lactobacillus jensenii, Lactobacillus crispatus, Lactobacillus gestatus, and Lactobacillus iners. Clinical studies have shown that after being implanted into the vagina, lactobacilli can adhere to and colonize vaginal epithelial cells, and produce lactic acid by decomposing glycogen to maintain the acidic environment of the vagina, while producing H 2 O 2 , bacteriocins, lysozymes and other substances inhibit the growth of pathogenic bacteria. Lactic acid bacteria refers to a general term for a class of non-spore, Gram-positive bacteria whose main product of fermentation of sugars is lactic acid. Most of them are essential bacteria in the human body and have important physiological functions. They are widely present in the human intestine. Lactic acid bacteria are facultative anaerobic bacteria and prefer to grow in an anaerobic state, but will not die due to contact with oxygen. The present invention can set monoclonal antibodies for STD items to be tested, monoclonal antibodies against lactobacilli and IgY antibodies on colored microspheres, thereby achieving simultaneous detection of multiple targets of microorganisms related to vaginitis and evaluating the health of the microecology. The detection process is simple, and the efficiency and reliability of the detection are improved.

[0034] Specifically, the sample pad is connected to the binding pad and pasted on the other side of the end where the binding pad is connected to the nitrocellulose membrane. The width of the overlapping area between the sample pad and the binding pad can be 3 mm. The absorbent pad is pasted on the other end of the nitrocellulose membrane away from the binding 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. This layout allows the sample to flow through the nitrocellulose membrane toward the absorbent pad after being added to the sample pad. In this process, substances such as colored microspheres in the binding pad react with the sample, thereby realizing the detection of the target substance in the sample, which will not be repeated here.

[0035] Specifically, determining the coefficient of rebound includes, The overlap area values ​​of the contour images of adjacent frames within a preset acquisition period are calculated, and the minimum value of the overlap area values ​​is determined as the rebound coefficient.

[0036] Specifically, the collected contour image can be preprocessed by graying and denoising, contours can be extracted using an edge algorithm, and the contour image can be converted into a binary image based on a pixel counting method to obtain the overlap area value, which will not be elaborated here.

[0037] See also Figure 2 As shown, it is a logic flow chart of determining whether there is a risk of abnormal bonding in the overlapping area according to an embodiment of the present invention. Determining whether there is a risk of abnormal bonding in the overlapping area includes: If the coefficient of resilience 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; If the coefficient of resilience 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; The bonding risk determination condition is that the rebound coefficient does not exceed a preset rebound coefficient reference value.

[0038] Specifically, the preset rebound coefficient reference value is the product of the area of ​​the overlapping area before rolling and the rebound factor. The rebound factor can be set by technical personnel in this field according to the preparation accuracy requirements of the test paper. The higher the accuracy requirement, the smaller the rebound factor. The value range of the rebound factor can be [0.7, 0.9]. Preferably, the rebound factor can be 0.8.

[0039] Specifically, 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. In the production process, the overlapping area can be found at an early stage. The abnormal fitting risk that may exist in the domain, since the contour images are continuously acquired and analyzed within the preset acquisition period, even the slightest change in fitting or abnormal situation can be detected, which helps to take timely measures for adjustment and improvement, and avoids discovering problems in subsequent production steps, thereby reducing material waste, improving production efficiency, and reducing production costs. By accurately determining the abnormal fitting risk, it can be ensured that the fitting quality of the overlapping area between the binding pad and 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, enhance the quality stability and reliability of the product, and further, realizes the rapid identification of the abnormal fitting risk tendency of the nitrocellulose membrane in the preparation process of the test strip, and improves the reliability of the preparation of the test strip.

[0040] Specifically, nitrocellulose membrane is a porous material with certain non-uniformity in pore size, pore distribution and fiber arrangement. These differences in microstructure lead to different water absorption capacity of different parts. After applying glue, the curing process of glue is usually affected by moisture. Areas with strong water absorption will absorb more water, which will delay the curing reaction of glue and cause the curing rate to slow down. In areas with weak water absorption, the glue is less affected by moisture and the curing rate is relatively fast. When the pasted binding pad is rolled, the binding pad and nitrocellulose membrane will be deformed by external force. Due to the uneven water absorption of nitrocellulose membrane, the curing rate of glue after applying glue is different. After the rolling external force disappears, the material will change due to the degree of curing. Different degrees of rebound are produced due to different curing rates. In the part with fast curing rate, the bonding structure formed by the glue is relatively stable and the rebound is small. In the part with slow curing rate, the bonding structure is not yet completely stable and there will be a large rebound. The overlapping area value of the contour images of adjacent frames 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. Conversely, the smaller the overlapping area, the greater the shape change and the greater the rebound. The minimum value of the overlapping area value is determined as the rebound coefficient, which can reflect the situation in which the overlapping area has the largest shape change in the entire rebound process. Furthermore, in the preparation process of the test paper, the abnormal risk tendency of the nitrocellulose membrane is quickly identified, and the reliability of the preparation of the test paper is improved.

[0041] Specifically, the process of determining the fit characterization parameters includes: A plurality of contour characterization values ​​are obtained on the contour image of the feature frame, and the variance of the contour characterization values ​​is determined as a fitting characterization parameter.

[0042] The feature frame is the last frame in a preset acquisition period, and the contour characterization value is a value in the width direction of the overlapping area.

[0043] See also Figure 3 As shown, it is a logic flow chart of determining the spraying abnormality category of the overlapping area according to an embodiment of the present invention. Determining the spraying abnormality category of the overlapping area includes: If the lamination 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; If the lamination 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; The first abnormality determination condition is that the fitting characterization parameter does not exceed a preset fitting characterization reference value.

[0044] Specifically, the preset fitting characterization reference value can be set by a technician in this field according to the accuracy requirement of the test paper preparation. The higher the accuracy requirement, the smaller the preset fitting characterization reference value. Preferably, the fitting characterization reference value can be 0.5.

[0045] Specifically, the present invention determines the contour characterization value according to 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 value in the width direction of the overlapping area in the contour image of the feature frame as the contour characterization value, it is possible to focus on the key features of the overlapping area in the final state after rolling, and calculate the variance of the contour characterization value as the fitting characterization parameter, so as to 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 means that the numerical value change in the width direction is small, and there is a problem of consistent overall fitting abnormality degree in the overlapping area, and it is 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 means that the value changes greatly in the width direction, and there is a problem of inconsistent overall fitting abnormality in the overlapping area. It is determined as the second spraying abnormality category, which is helpful for targeted analysis of different types of abnormalities and provides an accurate basis for subsequent adjustments. The present invention determines the contour characterization value through 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, in the preparation process of the test paper, it is realized that the risk tendency of fitting abnormality of the nitrocellulose membrane can be quickly identified, thereby improving the reliability of the preparation of the test paper.

[0046] Specifically, the spraying adjustment methods of the selected colored microspheres include: If the spraying abnormality category is the first spraying abnormality category, the spraying adjustment method of the selected colored microspheres is to adjust the spraying angle of the colored microspheres to the bonding pad according to the rebound coefficient; If the spraying abnormality category is the second spraying abnormality category, the spraying adjustment method of the selected colored microspheres is to turn on the vibration device of the plastic base plate during the spraying process, and adjust the vibration frequency of the vibration device according to the fitting characterization parameter.

[0047] Specifically, the vibration device can be an electromagnetic vibration device, which works on the principle of electromagnetic induction and generates an alternating magnetic field through an alternating current, so that the magnetic vibration component moves back and forth under the action of the magnetic field force, thereby generating vibration, which will not be elaborated here.

[0048] Specifically, the adjustment amount of the spraying angle is positively correlated with the rebound coefficient.

[0049] Specifically, a nozzle that can adjust the spraying angle by a motor drive can be used, and a microprocessor is used to control the motor to adjust the spraying angle. The direction of the nozzle perpendicular to the binding pad is marked as 0°, and the nozzle is rotated 20° toward the nitrocellulose membrane as the initial spraying angle. The spraying angle can range from [20, 60], and the interval unit is °.

[0050] Specifically, under the condition of the first spraying abnormality category, the present invention adjusts the spraying angle of the colored microspheres to the binding pad according to the rebound coefficient. It can be understood that when the binding pad and the nitrocellulose membrane are overlapped and pasted, there will be a slope with a certain height difference. The different glue curing rates in the overlapping area will generate an upward rebound force in the overlapping area, which increases the angle between the binding pad and the nitrocellulose membrane, thereby increasing the slope gradient. The microspheres are more likely to roll down the slope under the action of gravity and accumulate at the bottom of the slope, affecting the spraying uniformity of the colored microspheres. The first spraying abnormality category, that is, the angle of the slope is uniformly increased as a whole. Adjusting the spraying angle will change the initial velocity direction and force conditions of the colored microspheres when they are sprayed out. When the slope angle becomes larger, if it is still sprayed at the original angle, the microspheres are affected by gravity and the slope angle, and are more likely to roll down the slope quickly to the bottom and accumulate. Therefore, in the preparation process of the test paper, the risk tendency of abnormal bonding of the nitrocellulose membrane is quickly identified, the spraying method of the colored microspheres is adaptively adjusted, and the spraying uniformity of the colored microspheres and the reliability of the preparation of the test paper are improved.

[0051] Specifically, the vibration frequency is positively correlated with the fitting characterization parameter.

[0052] Specifically, the setting range of the vibration frequency may be [50, 150], and the interval unit is Hz.

[0053] Specifically, in the present invention, under the second spraying abnormality category condition, a vibration device for the plastic base plate is turned on during the spraying process, and the vibration frequency of the vibration device is adjusted according to the fitting characterization parameters. It can be understood that the second spraying abnormality category condition, that is, the angle change of the slope is different, and it is not uniformly increased as a whole. The slope with angle change differences will cause the colored microspheres to gather locally more easily, and the external force generated by the vibration can give the colored microspheres additional kinetic energy. This kinetic energy enables the microspheres to overcome the friction 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 area, and then roll to other areas of the slope, thereby reducing the accumulation amount at the bottom and making the microspheres on the slope The distribution of colored microspheres is more uniform. The slopes at different positions are different, and the distribution of colored microspheres will also be uneven. The vibration external force can make the microspheres readjust their positions according to the actual shape and slope changes of the slope. During the vibration process, the microspheres tend to move to areas with gentler slopes and larger spaces, thereby improving the overall distribution uniformity. In areas where the slope suddenly increases, the colored microspheres will move upward for a distance under the action of vibration to achieve a more balanced distribution state. Furthermore, in the preparation process of the test paper, the abnormal risk tendency 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 paper can be improved.

[0054] See also Figure 4 As shown, it is a schematic diagram of a test paper for rapid detection of microorganisms according to an embodiment of the present invention. A test paper for rapid detection of microorganisms according to the present invention comprises: A detection sample layer, which includes a sample pad 3 for filtering the sample solution and a binding pad 2 for fixing the colored microspheres; A test result display layer, which is connected to the test sample layer and includes a nitrocellulose membrane 1 for displaying the test result; The water absorption layer is connected to the detection result display layer and includes a water absorption pad 4 for absorbing excess sample solution.

[0055] Specifically, the colored microspheres are provided with monoclonal antibodies for detecting STD items of the microorganisms to be tested, and monoclonal antibodies against lactobacilli, wherein the monoclonal antibodies for STD items are one of the monoclonal antibodies of Gardnerella, monoclonal antibodies of gonococci, monoclonal antibodies of Trichomonas, monoclonal antibodies of Chlamydia, monoclonal antibodies of Mycoplasma, and monoclonal antibodies of Candida albicans.

[0056] Specifically, the semi-quantitative detection of Lactobacillus S-layer protein or N-protein can be used as an auxiliary diagnosis for microecological detection, which can characterize whether the microecological ecology is healthy or not. In the sample to be tested, if the lactobacillus is strongly positive and the content of the pathogen antigen to be tested is low, it indicates that the sample population is in a lactobacillus-dominated state and there is no BV indication; in the sample to be tested, if the 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 pathogen-dominated state and there is a BV indication; in the sample to be tested, if the lactobacillus is negative and the content of the pathogen antigen to be tested is obvious, it indicates that the sample population is in a pathogen-dominated state and there is a severe BV indication.

[0057] Exemplarily, a specific embodiment of vaginitis detection using Lactobacillus 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%, 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 bacterial vaginitis patient.

[0058] So far, the technical solutions of the present invention have been described in conjunction with the preferred embodiments shown in the accompanying 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 fall within the protection scope of the present invention.

[0059] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for preparing a test paper for rapid detection of microorganisms, characterized in that: include: Paste the nitrocellulose membrane required for the test paper on the plastic base plate, and paste the binding pad for fixing the colored microspheres to one end of the nitrocellulose membrane with a preset overlapping width; The pasted bonding pad is rolled, and contour images of overlapping areas of several frames are obtained within a preset acquisition period. The rebound coefficient is determined according to the comparison of contour images of different frames to determine whether there is a risk of abnormal bonding in the overlapping area; In response to the presence of a risk of abnormal lamination, a contour characterization value is determined according to a contour image of a feature frame within a preset acquisition period, and a lamination characterization parameter is determined according to a comparison of a plurality of contour characterization values ​​to determine a spraying abnormality category of the overlapping area; The spraying adjustment method of the colored microspheres is selected according to the spraying abnormality category, including: Adjusting the spraying angle of the colored microspheres onto the bonding pad according to the rebound coefficient; and, during the spraying process, turning on a vibration device for the plastic base plate, and adjusting the vibration frequency of the vibration device according to the bonding characterization parameter; The sample pad and the absorbent pad are pasted to the preset position of the plastic bottom plate, and connected to the nitrocellulose membrane and the binding pad to obtain the target test paper.

2. The method for preparing a test paper for rapid detection of microorganisms according to claim 1, characterized in that: Determining the coefficient of rebound includes, The overlap area values ​​of the contour images of adjacent frames within a preset acquisition period are calculated, and the minimum value of the overlap area values ​​is determined as the rebound coefficient.

3. The method for preparing a test paper for rapid detection of microorganisms according to claim 2, characterized in that: Determining whether there is a risk of abnormal bonding in the overlapping area includes: If the coefficient of resilience of the overlapping area meets the bonding risk determination condition, it is determined that the overlapping area has a bonding abnormality risk; If the coefficient of resilience 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; The bonding risk determination condition is that the rebound coefficient does not exceed a preset rebound coefficient reference value.

4. The method for preparing a test paper for rapid detection of microorganisms according to claim 3, characterized in that: The process of determining fit characterization parameters includes, Acquire a plurality of contour characterization values ​​on the contour image of the feature frame, and determine the variance of the contour characterization values ​​as a fitting characterization parameter; The feature frame is the last frame in a preset acquisition period, and the contour characterization value is a value in the width direction of the overlapping area.

5. The method for preparing a test paper for rapid detection of microorganisms according to claim 4, characterized in that: The types of spray anomalies that determine the overlap area include, If the lamination 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; If the lamination 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; The first abnormality determination condition is that the fitting characterization parameter does not exceed a preset fitting characterization reference value.

6. The method for preparing a test paper for rapid detection of microorganisms according to claim 5, characterized in that: The spraying adjustment methods of selected colored microspheres include: If the spraying abnormality category is the first spraying abnormality category, the spraying adjustment method of the selected colored microspheres is to adjust the spraying angle of the colored microspheres to the bonding pad according to the rebound coefficient; If the spraying abnormality category is the second spraying abnormality category, the spraying adjustment method of the selected colored microspheres is to turn on the vibration device of the plastic base plate during the spraying process, and adjust the vibration frequency of the vibration device according to the fitting characterization parameter.

7. The method for preparing a test paper for rapid detection of microorganisms according to claim 6, characterized in that: The adjustment amount of the spraying angle is positively correlated with the rebound coefficient.

8. The method for preparing a test paper for rapid detection of microorganisms according to claim 6, characterized in that: The vibration frequency is positively correlated with the fitting characterization parameter.

9. A test paper for rapid detection of microorganisms, using the method for preparing a test paper for rapid detection of microorganisms according to any one of claims 1 to 8, characterized in that: include: A detection sample layer, which includes a sample pad for filtering the sample solution and a binding pad for fixing the colored microspheres; A test result display layer, which is connected to the test sample layer and includes a nitrocellulose membrane for displaying the test result; The water absorption layer is connected to the detection result display layer and includes a water absorption pad for absorbing excess sample solution.

10. The test paper for rapid detection of microorganisms according to claim 9, characterized in that: The colored microspheres are provided with monoclonal antibodies for detecting STD items of microorganisms to be tested, and monoclonal antibodies against lactobacilli, wherein the monoclonal antibodies for STD items are one of the monoclonal antibodies of Gardnerella, monoclonal antibodies of gonococci, monoclonal antibodies of Trichomonas, monoclonal antibodies of Chlamydia, monoclonal antibodies of Mycoplasma, and monoclonal antibodies of Candida albicans.

Images