CN117741129A - Noninvasive oral mucosa tissue exudate protein level detection device - Google Patents
Noninvasive oral mucosa tissue exudate protein level detection device Download PDFInfo
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Abstract
The invention relates to the technical field of detection and sampling, in particular to a noninvasive device for detecting the protein level of oral mucosa tissue exudates, which comprises a negative pressure aspirator, wherein one side of an adsorption cavity is provided with two equidistant and parallel microporous layers to form an intermediate gap for placing adsorption materials; the other side of the adsorption cavity is connected with a pipeline with a bent angle; and a one-way valve is arranged at one end of the pipeline connected with the adsorption cavity; the effusion detection box comprises a shell, a sample injection port, an observation area, a supporting sheet and detection test paper; the sample main inlet is positioned at the top center of the upper surface of the shell; a supporting sheet extending horizontally is arranged below the sample injection opening, and a detection test paper is fixed on the supporting sheet. The invention can rapidly adsorb and acquire the exudates of the oral mucosa tissues, and reduce the pain and the wound of patients; the device combines the specific antigen-antibody reaction and layering display, clearly judges positive and negative, performs semi-quantitative analysis, and reflects the change of protein level before and after treatment in real time.
Description
Technical Field
The invention relates to the technical field of detection sampling, in particular to a noninvasive device for detecting the level of oral mucosa tissue exudates protein.
Background
The oral mucosa tissue fluid is naturally secreted by mucosa cells, contains bioactive molecules, cells, electrolytes, proteins and the like, and has a key meaning in a plurality of physiological and pathological states. The process of occurrence and development of diseases can be monitored at a molecular level by acquiring exudates from oral mucosa tissues, and potential biomarkers in the exudates can be used for making the oral mucosa tissues exudates a powerful tool for early diagnosis and disease progress monitoring. In order to ensure the accuracy of diagnosis, strict requirements are placed on the collection amount and quality of the exudates, namely, the stability and consistency of the characteristics of the collected fluids are required to be ensured, meanwhile, the interference of mixed saliva and the degradation of various enzymes in the saliva on sample proteins are required to be avoided, and the sufficient collection amount is required to be ensured to meet the minimum requirement of detection.
The existing collection means, such as a scraping method, a cotton suction method and the like, are mixed with saliva components in collected tissue exudates, and the saliva mainly comes from exocrine glands such as parotid glands, submandibular glands and sublingual glands and is greatly affected by oral flora and collection time, so that the actual conditions in mucous membrane tissues cannot be truly and effectively reflected; and these means have some uncertainty in the time of collection, the location and the amount of fluid collected, which can lead to deviations in the analysis of the biomarkers. In addition, in disease diagnosis and progression monitoring, semi-quantitative/qualitative data can provide a relatively complete detection analysis for biomarkers, contributing to a more accurate assessment of health status, making the assessment more scientific and comparable, whereas most collection means ignore the extraction of amounts, or lack stability, failing to ensure stability and consistency of oral mucosal tissue exudates, presenting challenges for subsequent semi-quantitative analysis.
Therefore, considering the characteristics of oral mucosa tissue exudates and the requirements of collection, the same noninvasive extraction of the oral mucosa tissue exudates is required, the oral mucosa is prevented from being damaged again, the collection efficiency and accuracy are improved, and stable and accurate detection data are provided for the semi-quantitative analysis of the subsequent protein level.
Disclosure of Invention
The invention aims at solving the technical problems and provides a noninvasive device for detecting the level of the exudates protein of oral mucosa tissues, which can be realized by the following technical scheme:
the invention provides a noninvasive device for detecting the protein level of oral mucosa tissue exudates, which comprises a negative pressure aspirator and an exudates detection box,
the negative pressure aspirator comprises an adsorption cavity, a microporous layer, an adsorption material and a pipeline; two equidistant and parallel microporous layers are arranged on one side of the adsorption cavity to form an intermediate gap for placing adsorption materials; the pipeline is provided with a bent angle part, so that the negative pressure suction device can conveniently place the adsorption cavity at a position required to be collected through the bent angle part, one end of the pipeline is connected with a negative pressure device through a hose, and the other side of the adsorption cavity is connected with the pipeline; the one-way valve is arranged at the end of the pipeline connected with the adsorption cavity, so that the stability of the adsorption cavity during collection is ensured, and the negative pressure reverse flow is avoided;
the effusion detection box is made of hard materials and comprises a shell, a sample injection opening, an observation area, a supporting sheet and detection test paper; the sample main inlet is positioned at the top center of the upper surface of the shell and is used for receiving preservation solution containing the exudates; the observation area is positioned in the middle section of the upper surface of the shell and between the top and the bottom; a supporting plate extending horizontally is arranged below the sample injection opening, and detection test paper is fixed on the supporting plate and comprises a quality control strip and a detection strip which are both positioned in the observation area.
Further, the microporous layer has a width in the range of 3 to 7 mm. The pore width of the microporous layer ranges from 200 to 500 microns.
Further, the included angle of the bent angle of the pipeline is between 120 degrees and 150 degrees.
Further, the absorbent material comprises sampling absorbent paper or porous spongy high molecular polymer.
Further, the test strip comprises a quality control strip comprising a general quality control strip and a saliva protein control strip. Typically, the quality control strip detects proteins in tissue exudates, while the saliva quality control strip detects proteins in saliva.
Further, the test strip comprises at least 4 color strips for detecting the type or concentration of the specific protein, respectively.
Compared with the prior art, the invention has at least one of the following technical effects:
the oral mucosa tissue exudates protein level device designed by utilizing the negative pressure principle can (1) obtain oral mucosa tissue exudates noninvasively, reduce the injury to patients, can be implemented for multiple times, and is beneficial to developing disease monitoring; (2) The tissue part to be used for collecting exudates can be selected according to actual needs, so that the accurate and true reaction of diseases or disease states of specific tissue parts is facilitated; (3) The interference of saliva microenvironment and oral flora on a sample to be acquired is reduced to the greatest extent, and the reliability of a detection result is improved; (4) The device combines the specific antigen-antibody reaction and layering display, not only clearly judges positive and negative, but also can perform semi-quantitative analysis and reflect the change of the biomarker protein level before and after treatment in real time; (5) Multiple proteins and cytokines can be detected simultaneously by a single sampling. Compared with the traditional method, the device is simple, efficient and stable, and the detection result is real and stable. The device is not only limited to hospitals, but also can be used for preliminary screening or tracking observation in communities, pharmacies and the like, and the possibility of early discovery and intervention of oral diseases is greatly improved.
Drawings
In order to more clearly illustrate the technical solution of the embodiments of the present invention, the following description will briefly explain the drawings that are required to be used in the description of the embodiments:
FIG. 1 is a schematic diagram of the structure of a negative pressure aspirator and an exudate detection box of the noninvasive oral mucosa tissue exudate protein level detection device of the present invention;
FIG. 2 is a schematic diagram of a non-invasive negative pressure aspirator without adsorbing material for detecting oral mucosa tissue exudates protein level device according to the present invention;
FIG. 3 is a schematic diagram of the operation of the quality control strip and the test strip in the exudate test cassette of the present invention.
Description of the reference numerals
1: a pipe; 2: bending angles; 3: a one-way valve;
4: a suction head; 401: an adsorption chamber; 402: a microporous layer A;403: an adsorption material; 404: an intermediate gap; 405: a microporous layer B;
5: a housing; 6: a sample injection port; 7: an observation area; 8: and (5) detecting test paper.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be further described in detail below with reference to the accompanying drawings and detailed description. It should be understood that the detailed description is presented by way of example only and is not intended to limit the invention.
Example 1
The embodiment provides a noninvasive device for detecting the protein level of oral mucosa tissue exudates, which comprises a negative pressure aspirator and an exudates detection box,
referring to fig. 1 and 2, a suction head 4 of a negative pressure suction apparatus includes an adsorption cavity 401, a microporous layer, an adsorption material 403 and a pipe 1; two equidistant and parallel microporous layers are arranged on one side of the adsorption cavity 401 to form an intermediate gap 404 for placing an adsorption material 403; the pipeline 1 is provided with a bent angle part, so that the negative pressure suction device can conveniently place the adsorption cavity at a position to be collected through the bent angle part, one end of the pipeline is connected with a negative pressure device through a hose, and the other end of the adsorption cavity 401 is connected with the other end of the pipeline 1 with the bent angle 2; and the one end of the pipeline 1 connected with the adsorption cavity 401 is provided with the one-way valve 3, so that the stability of the adsorption cavity during collection is ensured, and the negative pressure reverse flow is avoided.
Two layers of microporous layers are sequentially arranged inside one end of the adsorption cavity 401, and the width range of the microporous layers is 3-7 mm; the pore width of the microporous layer ranges from 200 to 500 microns, and the microporous layer design ensures that liquid enters and is absorbed by the adsorbent material 403; the microporous layer is sandwiched by an adsorbent material 403. The joint of the other end of the adsorption cavity 401 and the pipeline 1 is provided with a one-way valve 3, the pipeline 1 is provided with a bent angle 2, and the included angle of the bent angle 2 of the pipeline 1 is between 120 and 150 degrees; the negative pressure aspirator is convenient to be placed at the position required to be collected;
the negative pressure aspirator is made of hard materials, and the hard materials can be PTE or ABS and other materials; as shown in fig. 2, the microporous layer a 402 is a single layer, the outer microporous layer B405 is connected with the adsorption cavity 401 in a connection manner including spiral connection, quick snap connection, etc., the outer microporous layer B405 and the adsorption cavity 401 are provided with threads matching each other, and mechanical fixation can be achieved by screwing in the adsorption cavity, in addition, the connection manner can also adopt a quick snap mode, a tiny snap or button is arranged on the outer ring wall of the outer microporous layer B405, and a matched clamping position is arranged on the inner side or the outer side of the adsorption cavity 401, so that the outer microporous layer B405 can be fixed on the adsorption cavity 401 or conveniently removed by simply pressing the snap or button, and the two mentioned methods not only ensure stable connection between the microporous layer B405 and the adsorption cavity 401, but also make the replacement process of the microporous layer B405 convenient and simple.
The adsorption material can be paper, porous spongy high molecular polymer or cotton piece, and the like, and the quantitative method provides accurate, reliable and comparable data for us when detection and research are carried out, so that the accuracy and reproducibility of the result are ensured, and especially in the scene involving liquid volume and biological molecule concentration, different volumes or concentrations can cause obvious effects, so that the collected exudates of the oral mucosa need to be accurately quantified. In order to ensure that the adsorbent material is the same material and uniform texture for each sample, the maximum saturated water absorption should be the same in this case, which can be controlled to be 5 μl or 10 μl. These adsorbent materials have high water absorbability and biomolecule retention capacity. According to the design of the study, the diameter and the volume of the adsorption material contained in the negative pressure aspirator should be completely consistent. In the case of a relatively standard operation, it should be that the whole piece of absorbent material is completely saturated with liquid during sampling, while the suction apparatus is removed, a portion of the excess liquid should be able to flow out of it, i.e. it means that the material has been completely saturated and no more liquid can be absorbed.
In addition, the application of negative pressure suction to the oral mucosa is based on the unique properties of the oral mucosa, which is clearly different from the skin. The oral mucosa and skin, although both are external, covered tissues of the human body, differ in structure, function and manner of responding to external stimuli. The following are some key differences between the two and their effect on negative pressure suction: from the difference of tissue structure, the oral mucosa is thinner than the skin, and the epithelial layer and the basal layer are thinner and transparent. Whereas the epidermis layer of the skin is thicker, especially the stratum corneum is more pronounced. This means that for negative pressure applications, the oral mucosa is more easily permeable to liquids. From the difference between blood flow and interstitial fluid: the blood vessels in the oral mucosa are rich and are closer to the surface of the mucosa, and the tissue fluid content is relatively high. This makes it easier for interstitial fluid in the mucosa to be drawn out when we apply negative pressure. On the oral mucosa, due to the special tissue characteristics and higher repairing capability than skin, the application of moderate negative pressure is not easy to cause obvious damage.
Thus, based on the above analysis, the application of negative pressure suction on the oral mucosa does have its uniqueness and advantages, fundamentally different from the application on the skin. These differences should indeed be taken into account and the advantages of the oral mucosa fully utilized while ensuring safety and effectiveness of the procedure when conducting experiments or applications.
Referring to fig. 2, the exudate detection box includes a housing 5, a sample injection port 6, an observation area 7, a support sheet and a detection test paper 8; the sample main inlet is positioned at the top center of the upper surface of the shell 5 and is used for receiving preservation solution containing exudates; the observation area 7 is positioned in the middle section of the upper surface of the shell 5 and is positioned between the top and the bottom; a supporting plate extending horizontally is arranged below the sample injection opening 6, and a detection test paper 8 is fixed on the supporting plate, and the detection test paper 8 comprises a quality control strip and a detection strip which are both positioned in the observation area. The quality control strips included in the test strip 8 include a general quality control strip (C1) and a saliva quality control strip (C2). Typically, the control strip (C1) detects proteins in tissue exudates, while the saliva control strip (C2) detects proteins in saliva. The detection strip at least comprises 4 color strips, and the protein concentration is detected.
The working principle of the test strip 8 is antigen-antibody reaction and enzyme-catalyzed chromogenic reaction (similar to enzyme-linked immunosorbent assay enzyme linked immunosorbent assay, ELISA). The protein polypeptide in the exudates is used as antigen, namely the detected object, the corresponding specific antibody is preset on the strip, and when the proteins in the exudates infiltrate the strip part, the color can be displayed on the strip part through antigen-antibody reaction and enzyme catalysis color reaction.
Referring to fig. 3, in the quality inspection area, the C1 and C2 bands are control bands, i.e., quality control bands. The C1 band is intended to be a protein polypeptide which is stable and conservatively present in tissues, such as beta actin (beta-actin), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), collagen (collagen), etc. The C2 band is intended to be selected from protein polypeptides which exist stably in saliva, such as sialoadhesin, etc. The color development of the C1 band represents that the tested sample contains exudates and substances (including proteins) from tissue sources, and the color development of the C2 band represents that the tested sample contains fluids and substances (including proteins) from saliva sources. For detecting tissue exudates, the C1 band should be colored and the C2 band should not be colored, i.e. the sample to be detected contains only exudates and substances from tissue sources, but not fluids and substances from saliva, so that interference of saliva components can be avoided.
T1-T4 are detection bands on which specific antibodies and enzyme-catalyzed chromogenic reagent systems have been pre-placed. According to the design of presetting antibodies with different concentrations, the function of semi-quantitatively detecting the content of the corresponding antigen is achieved. Taking Granzyme B (GZB, granzyme B) as an example, the GZB detectable concentration of the T1 band (i.e., the concentration of the preset antibody) was set at 10pg/mL; the concentration of the preset antibody is increased by the T2 strip according to the proportion of 5 times, namely the detectable concentration of the GZB is 50pg/mL; similarly, the detectable concentration of the T3 band is 250pg/mL and the T4 band is 1250pg/mL.
Example 2
Unlike the exudates assay cartridge for detecting protein concentration mentioned in example 1, the design objective of example 2 is to provide a multifunctional antigen assay cartridge that can qualitatively detect different types of antigens.
According to the design of presetting different types of antibodies, the function of qualitatively detecting the corresponding antigens is achieved. On the test strip 8, a quality test strip and a test strip are included.
The design of a general quality control strip (C1) and a saliva quality control strip (C2) in the quality control strip is unchanged, and different antibodies and enzyme catalysis color development systems are respectively preset on the T1-T4 strips, wherein the special design comprises type I interferon, various types of interleukins, tissue growth factors and the like. Each band contains a specific enzyme-catalyzed color development system that produces a color change upon binding to the antigen of interest, thereby enabling the user to visually read the results
The operation steps are as follows:
(1) The sample is added drop wise to the sample well of the cartridge.
(2) Waiting for a recommended period of time (e.g., 10 minutes), the color change on the test strip is observed.
(3) The result is read. If C1 develops color and C2 does not develop color, the detection box is indicated to work normally. The color change of the T1-T4 bands then indicates the presence of the corresponding antigen in the sample.
Because the detection box can detect multiple antigens simultaneously, the detection box can be suitable for occasions needing rapid and wide screening, such as screening in the case of outbreak of pandemic or in the hospital environment needing rapid diagnosis. In addition, the cartridge can be used in research to rapidly compare the presence of various biomolecules in different samples.
Example 3
Based on the same inventive concept, the invention also provides a noninvasive method for detecting the level of the oral mucosa tissue exudates protein, which is applied to the device for detecting the level of the oral mucosa tissue exudates protein, and is characterized by referring to figures 1 and 2, comprising the following steps,
step S1, ordering a patient to rinse with normal saline, cleaning an oral cavity, determining a part to be collected, gently sucking saliva on the oral mucosa surface of the part with cotton balls or gauze, and taking care of moisture isolation;
step S2, connecting a pipeline 1 interface of the negative pressure aspirator to a negative pressure device, such as air extraction equipment, and tightly attaching a microporous layer B405 on the outer side of the negative pressure aspirator to oral mucosa of a part to be collected;
step S2, opening the air extraction equipment to enable exudates from the oral mucosa tissues to be absorbed into the material absorbing material 403 in the middle gap 404 through the outer microporous layer B405; when the water absorption reaches saturation or reaches a preset sampling amount, or the liquid is observed to start flowing to the one-way valve 3, the air extraction equipment is closed, the material adsorption material layer is taken down, the material adsorption material layer is placed into the preservation liquid, and the sufficient mixing of the effusion and the preservation liquid is ensured.
Step S4, sample liquid in a preset range is taken from the preservation liquid, dripped into the sample injection port 6, and enters the detection test paper 8 through the sample injection port and starts to infiltrate and diffuse;
and S5, in the infiltration process of the sample liquid, judging whether the source of the sample liquid is from tissue infiltration liquid or mixed with protein in saliva in the quality control strip, so as to ensure the accuracy of the sample liquid. When the sample liquid infiltrates into the detection strip, specific proteins in the tissue infiltration liquid can be captured by the corresponding antibodies, so as to generate a color development strip; in qualitative detection, observing the appearance position of the chromogenic band, and determining whether the target protein or the corresponding antigen thereof exists; in the semi-quantitative detection, the relative concentration of the target protein after the dilution in equal proportion is determined according to the number of color bands.
Although the present invention has been described in terms of the preferred embodiments, it is not intended to be limited to the embodiments, and any person skilled in the art can make any possible variations and modifications to the technical solution of the present invention by using the methods and technical matters disclosed above without departing from the spirit and scope of the present invention, so any simple modifications, equivalent variations and modifications to the embodiments described above according to the technical matters of the present invention are within the scope of the technical matters of the present invention.
Claims (6)
1. A noninvasive device for detecting the protein level of oral mucosa tissue exudates is characterized by comprising a negative pressure aspirator and a exudates detection box,
the negative pressure aspirator comprises an adsorption cavity, a microporous layer, an adsorption material and a pipeline; two equidistant and parallel microporous layers are arranged on one side of the adsorption cavity to form an intermediate gap for placing the adsorption material; the pipeline is provided with a bent angle part, so that the negative pressure suction device can conveniently place the adsorption cavity at a position required to be collected through the bent angle part, one end of the pipeline is connected with a negative pressure device through a hose, and the other end of the pipeline is connected with the other side of the adsorption cavity; the one-way valve is arranged at the end, connected with the adsorption cavity, of the pipeline, so that the stability of the adsorption cavity during collection is ensured, and negative pressure reverse flow is avoided;
the effusion detection box comprises a shell, a sample injection port, an observation area, a supporting sheet and detection test paper; the sample injection port is positioned at the center of the top of the upper surface of the shell and is used for receiving preservation solution containing the effusion; the observation area is positioned in the middle section of the upper surface of the shell and is positioned between the top and the bottom; the sample filling opening is characterized in that a supporting sheet extending horizontally is arranged below the sample filling opening, and the supporting sheet is fixedly provided with detection test paper which comprises a quality control strip and a detection strip and is positioned in the observation area.
2. The device for detecting the level of oral mucosal tissue exudates protein according to claim 1, wherein the width of said microporous layer is in the range of 3 to 7 mm; the microporous layer has a pore width in the range of 200 to 500 microns.
3. The device for detecting the level of transudate protein in an oral mucosa tissue according to claim 2, wherein the included angle of said bent angle of said tube is in the range of 120 ° to 150 °.
4. The device for detecting the level of exudate proteins in oral mucosal tissue of claim 1, wherein the adsorbent material comprises a sampling absorbent paper or a porous sponge-like polymer.
5. The device for detecting oral mucosal tissue exudate protein level as in claim 1, wherein the quality control strip comprised by the test strip comprises a general quality control strip and a saliva protein control strip; the general quality control strip detects proteins in tissue exudates, while the saliva quality control strip detects proteins in saliva.
6. The device for detecting the level of exudate proteins in oral mucosal tissue according to claim 5, wherein the detection strip comprises at least 4 color strips for detecting the type or concentration of the specific protein, respectively.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311672600.8A CN117741129A (en) | 2023-12-07 | 2023-12-07 | Noninvasive oral mucosa tissue exudate protein level detection device |
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| CN202311672600.8A CN117741129A (en) | 2023-12-07 | 2023-12-07 | Noninvasive oral mucosa tissue exudate protein level detection device |
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| CN117741129A true CN117741129A (en) | 2024-03-22 |
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| CN202311672600.8A Pending CN117741129A (en) | 2023-12-07 | 2023-12-07 | Noninvasive oral mucosa tissue exudate protein level detection device |
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