CN116196042B

CN116196042B - Periodontitis detection kit and application thereof

Info

Publication number: CN116196042B
Application number: CN202310080995.6A
Authority: CN
Inventors: 宋锦璘; 张曦木; 神华尉; 袁徐蕾
Original assignee: Stomatological Hospital of Chongqing Medical University
Current assignee: Stomatological Hospital of Chongqing Medical University
Priority date: 2023-02-08
Filing date: 2023-02-08
Publication date: 2023-11-03
Anticipated expiration: 2043-02-08
Also published as: CN116196042A

Abstract

The invention relates to a periodontitis detection kit and application thereof, wherein the kit comprises: the extraction device for extracting the gingival crevicular fluid sample comprises an extraction tube, wherein the extraction tube comprises an extraction end, one side of the extraction tube, which is close to the extraction end, is provided with a capillary channel, and an air pressure balance tube is arranged on the extraction tube; a mixing chamber having a receiving space for receiving a sample, the receiving space being in communication with the outlet end of the extraction tube; the first end of the pushing part can extend into the extraction tube from the outlet end, and the second end of the pushing part penetrates through the accommodating space and extends out of a first opening arranged on the accommodating space; when the pushing part moves along the direction close to the outlet end, the pushing part can act on the air in the extraction tube to drive the sample in the capillary channel to flow along the first direction. The kit is simple to operate, convenient for long-time storage and transportation of gingival crevicular fluid, and well suitable for rapid periodontitis detection aiming at large-scale people.

Description

Periodontitis detection kit and application thereof

Technical Field

The invention relates to the field of periodontitis detection, in particular to a periodontitis detection kit and application thereof.

Background

Periodontitis is one of three diseases of the oral cavity, and is also the sixth most common disease of human beings, and the global incidence rate is up to 45% -50%. The fourth national oral health epidemiological investigation report shows that the periodontal health rate of the population in China is less than 10 percent. Typical symptoms of periodontitis are destruction of tooth supporting tissue, including gingival bleeding, gingival atrophy, alveolar bone resorption, and even loosening and loss of teeth. Periodontitis is reported to be a major cause of loss of teeth in adults. Periodontitis severely impairs the chewing function of a patient, not only limits his food choice, affects aesthetics, but also has a great negative impact on the physical and mental health and quality of life of the patient. A number of reports have demonstrated that periodontitis is associated with a number of systemic diseases, such as cardiovascular and cerebrovascular diseases, diabetes, digestive tract cancers, etc. In addition, the course of periodontitis can be as long as decades, with a significant burden on individuals and society. Thus, early and accurate diagnosis of periodontitis is of paramount importance from the oral as well as general health perspective.

The clinical diagnosis of periodontitis is currently mainly dependent on the clinical examination of the dentist at his chair and the imaging examination. First, the dentist will ask the patient for medical history, such as systemic, oral, periodontal and oral hygiene habits, as well as family history. Subsequently, the dentist will conduct an examination of the periodontal tissue within the mouth of the patient, for example by plaque staining experiments and routine visual judgment, of the patient's oral hygiene (e.g. gingival index, bleeding index, etc.); judging whether the teeth of the patient have periodontal pockets, bleeding, bifurcation lesions and the like through periodontal probing; checking the degree of loosening of teeth by using dental forceps, checking whether there is an occlusal disturbance by using bite paper, etc.; in addition, the alveolar bone absorption can be visually displayed through X-ray images. As can be seen from the above, the above examination is not only time-consuming and requires a high degree of fit for the patient, but also the diagnosis results are subjective and are not conducive to objective assessment of the patient's state of development of periodontitis (e.g., most periodontitis patients are not at a fixed medical location, and even at the same medical location, the dentists who visit each time may be different).

Although there are some periodontal auxiliary examination methods (such as pressure sensitive probes, dental instruments, etc.), these auxiliary examinations are cumbersome and time-consuming in many steps, and do not facilitate the rapid completion of the examination by the dentist at the chair, and are not commonly used.

In addition, there are periodontal auxiliary examination methods for gingival crevicular fluid detection by collecting gingival crevicular fluid. However, the existing collection method has the defects of complicated steps or easiness in damaging gums of patients. At present, a gingival crevicular fluid collection method mainly comprises the following steps: gingival sulcus irrigation, capillary, filter paper strip, moisture-absorbing paper, etc. Among them, the gingival crevicular lavage method cannot determine the actual extraction amount of gingival crevicular fluid, and the diagnostic significance of the collected gingival crevicular lavage fluid is still to be clarified, so that the method has a certain limitation in practical application. The traditional capillary method can collect a large amount of gingival crevicular fluid, but has low extraction efficiency and poor collection effect stability. The filtering paper tape method is harmless, simple and convenient to operate, can quantitatively study the total amount of gingival crevicular fluid and biochemical components in the gingival crevicular fluid, and has relatively wide application. However, the sample obtained by the filter paper strip method needs to be subjected to operations such as dilution, elution and the like, so that the complex operation is performed, and the biological tissue characteristics in gingival crevicular fluid can be damaged, thereby influencing the subsequent diagnosis; and the gingival crevicular fluid content that the filter paper strip method can extract is less, need repeated sampling many times. The moisture-absorbing paper process also has similar drawbacks to the filter paper strip process. Among them, in order to solve the problem of small extraction amount of gingival crevicular fluid, chinese patent publication No. CN208769830U discloses a gingival crevicular fluid collecting device designed based on a moisture-absorbing paper method. Although the gingival crevicular fluid collecting device can increase the absorption amount of the gingival crevicular fluid to a certain extent under the action of the water absorption ball, when the gingival crevicular fluid is required to be taken out for periodontitis detection, the water absorption ball and the moisture absorption paper can not be prevented from remaining to absorb or adsorb a certain amount of gingival crevicular fluid. In other words, the water-absorbent ball and the moisture-absorbent paper inevitably cause a certain loss of gingival crevicular fluid.

Disclosure of Invention

The invention aims to provide a periodontitis detection kit. In order to solve the technical problems, the invention adopts the following technical scheme:

a periodontitis detection kit comprising:

an extraction device for extracting a gingival crevicular fluid sample, the extraction device comprising:

an extraction tube comprising an extraction end and an outlet end disposed on a side distal from the extraction end, wherein the extraction end is sized and shaped to extend between a tooth and an adjacent gum, the extraction tube is disposed as a capillary channel on a side proximal to the extraction end, and at least one air pressure equalization tube is disposed on the extraction tube;

a mixing chamber comprising a receiving space for receiving the sample, the receiving space being in communication with the outlet end and the receiving space being provided with a first opening at a location corresponding to the outlet end;

a pushing part, a first end of which can extend into the extraction tube from the outlet end, and a second end of which penetrates through the accommodating space and extends out from the first opening; when the pushing part moves along a first direction, the action of the pushing part on the air in the extraction pipe can drive the sample in the capillary channel to flow along the first direction, wherein the first direction is the direction from the extraction end to the outlet end.

In some embodiments, the extraction tube is provided with at least one section of graduation line for reading the extracted quantity of the sample.

In some embodiments, the extraction tube is provided with frangible structures at a side proximate the mixing chamber.

In some embodiments, the extraction end is rounded.

In some embodiments, the first end has an outer diameter dimension that is greater than an outer diameter dimension of the second end, and correspondingly, the outlet end has an inner diameter dimension that is greater than an inner diameter dimension of the first opening.

In some embodiments, the pushing portion is made of a rubber material.

In some embodiments, the receiving space is provided with a liquid outlet and the liquid outlet is provided with a sealed opening.

In some embodiments, the extraction tube comprises a fixed tube fixedly connected with the accommodating space, and a movable tube with one end detachably connected with the fixed tube, and the other end of the movable tube is the extraction end.

In some embodiments, further comprising: a test card, and the test card comprises:

a body; the sample adding port is arranged on the body, and an opening of the sample adding port is arranged on the first surface of the body;

The first detection areas are communicated with the sample adding port through a second channel.

The second aspect of the present invention provides a periodontitis detection kit, comprising:

a first storage cavity, wherein a first storage space for accommodating the sample is formed in the first storage cavity, the first storage cavity further comprises a pushing part, a first end of the pushing part is positioned in the first storage cavity, and a second end of the pushing part extends to the outside of the first storage cavity;

an extraction tube comprising an extraction end and an outlet end disposed on a side remote from the extraction end, wherein the outlet end is in communication with the first storage space, the extraction end is sized and shaped to extend between a tooth and an adjacent gum, and a side of the extraction tube proximate the extraction end is configured as a capillary channel;

the second storage cavity is internally provided with a second storage space for containing liquid, the second storage space is communicated with the first storage space through a first channel, at least one air pressure balancing port is also arranged in the second storage space, and one side of the first channel, which is close to the second storage space, is provided with a capillary structure;

When the pushing part moves along the direction far away from the first storage cavity, the action of the pushing part on the air in the extraction device can drive the sample in the capillary channel and the liquid in the second storage cavity to flow towards the direction close to the first storage cavity.

In some embodiments, the extraction tube includes a fixed tube fixedly connected to the first storage space, and a movable tube having one end detachably connected to the fixed tube, and the other end of the movable tube is the extraction end.

In some embodiments, the extraction end is rounded.

In some embodiments, the first storage space is provided with a second opening for balancing air pressure, and may also be used for draining the liquid in the first storage space.

In some embodiments, the first end has an outer diameter dimension that is greater than an outer diameter dimension of the second end.

In some embodiments, the pushing portion is made of a rubber material.

In some embodiments, further comprising: a test card, and the test card comprises: a body; the sample adding port is arranged on the body, and an opening of the sample adding port is arranged on the first surface of the body; the first detection areas are communicated with the sample adding port through a second channel.

The invention also provides an application of the kit in evaluating periodontitis of a subject, wherein the first detection areas in the detection card are respectively pre-stored with corresponding reagents, and the reagents comprise: one or more of an aptamer specifically recognizing human hemoglobin, an aptamer specifically recognizing Porphyromonas gingivalis, an aptamer specifically recognizing matrix metalloproteinase-8, an aptamer specifically recognizing interleukin-6, an aptamer specifically recognizing interleukin-1β, an aptamer specifically recognizing 8-hydroxydeoxyguanosine, an aptamer specifically recognizing malondialdehyde, and an aptamer specifically recognizing gingival proteinase K.

The beneficial technical effects are as follows:

the gingival crevicular fluid extraction device provided by the embodiment of the invention is particularly suitable for rapid initial screening of oral health of a subject. The extraction device adopts a mixed preservation mode for gingival crevicular fluid and diluent, so that the necessary extraction amount of gingival crevicular fluid is effectively reduced, the sampling efficiency of periodontitis can be improved, and gum damage to a subject in the sampling process can be reduced or avoided. Meanwhile, the extraction device can be used for stably pre-storing the diluent, so that a user can conveniently and rapidly realize the mixed storage operation of gingival crevicular fluid and diluent only through a single extraction device in the actual sampling process. In summary, the user can smoothly and sequentially perform the steps of sampling, reading, mixing, etc. on a single extraction device, so as to efficiently complete the sampling work.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. Like elements or portions are generally identified by like reference numerals throughout the several figures. In the drawings, elements or portions thereof are not necessarily drawn to scale. It will be apparent to those of ordinary skill in the art that the drawings in the following description are of some embodiments of the invention and that other drawings may be derived from them without undue effort.

Fig. 1 is a schematic structural view of an extraction device in a third exemplary embodiment of the present invention;

FIG. 2 is a schematic view showing the internal structure of the extracting apparatus shown in FIG. 1;

FIG. 3 is a schematic view showing the internal structure of a mixing chamber of the extraction device shown in FIG. 1;

FIG. 4 is a schematic view of the independent preservation unit of the extraction device of FIG. 1 after the gingival crevicular fluid has been extracted;

FIG. 5 is a schematic view of the extraction device of FIG. 1 in use;

fig. 6 is a schematic structural view of an extracting device in a fourth exemplary embodiment of the present invention;

FIG. 7 is a first schematic view showing the internal structure of the extracting apparatus shown in FIG. 6;

FIG. 8 is a second schematic view of the internal structure of the extraction device shown in FIG. 6;

FIG. 9 is a schematic view of the extraction device of FIG. 6 in use;

FIG. 10a is a first schematic structural view of the reverse side of a test card in an exemplary embodiment of the present invention;

FIG. 10b is a schematic diagram of the front structure of a test card in an exemplary embodiment of the invention;

FIG. 10c is a second schematic structural view of the opposite side of the test card in an exemplary embodiment of the present invention;

FIG. 11 is a schematic diagram of a test card and test equipment used with the test card in an exemplary embodiment of the invention;

FIG. 12 is a first schematic view of the plug-in mating of the test card of FIG. 11 with a test instrument;

fig. 13 is a second schematic view of the mating of the test card of fig. 11 with a test instrument.

Wherein 10 is an extraction pipe, 11 is a capillary channel, 12 is an extraction end, 13 is an air pressure balance pipe, 14 is a scale mark, and 15 is a breaking structure; 20 is a mixing cavity, 21 is a containing space, and 22 is a liquid outlet; 30 is a pushing part, 31 is a first end, and 32 is a second end; 40 is a first storage cavity, 41 is a second opening, and 42 is a first storage space; 50 is the second storage chamber, 51 is the atmospheric pressure balance mouth, and 52 is the second storage space, and 53 is first passageway, and 60 is the body, and 61 is the application of sample mouth, and 62 is first detection zone, and 63 is the second passageway.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. It will be apparent that the described embodiments are some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

The terms "upper," "lower," "inner," "outer," "front," "rear," "one end," "the other end," and the like herein refer to an orientation or positional relationship based on that shown in the drawings, merely for convenience of description and to simplify the description, and do not denote or imply that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus should not be construed as limiting the invention. Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.

The terms "mounted," "configured to," "connected," and the like, herein, are to be construed broadly as, for example, "connected," whether fixedly, detachably, or integrally connected, unless otherwise specifically defined and limited; the two components can be mechanically connected, can be directly connected or can be indirectly connected through an intermediate medium, and can be communicated with each other. The specific meaning of the above terms in the present invention will be understood in specific cases by those of ordinary skill in the art.

Herein, "and/or" includes any and all combinations of one or more of the associated listed items.

Herein, "plurality" means two or more, i.e., it includes two, three, four, five, etc.

It should be noted that, in this document, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising one … …" does not exclude the presence of other like elements in a process, method, article, or apparatus that comprises the element.

As used in this specification, the term "about" is typically expressed as +/-5% of the value, more typically +/-4% of the value, more typically +/-3% of the value, more typically +/-2% of the value, even more typically +/-1% of the value, and even more typically +/-0.5% of the value.

In this specification, certain embodiments may be disclosed in a format that is within a certain range. It should be appreciated that such a description of "within a certain range" is merely for convenience and brevity and should not be construed as a inflexible limitation on the disclosed ranges. Accordingly, the description of a range should be considered to have specifically disclosed all possible sub-ranges and individual numerical values within that range. For example, the description of ranges 1-6 should be considered as having specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as individual numbers within such ranges, e.g., 1,2,3,4,5, and 6. The above rule applies regardless of the breadth of the range.

The present invention provides a marker set for assessing the periodontitis status of a subject, the marker set comprising: a) The amount of gingival crevicular fluid; b) The pH of gingival crevicular fluid; c) Gingival crevicular fluid hemoglobin.

In some embodiments, the set of markers further comprises a porphyromonas gingivalis related marker comprising an amount of porphyromonas gingivalis and a gingival proteinase K (KGP).

In some embodiments, the set of markers further includes a periodontitis-related biochemical marker including matrix metalloproteinase-8 (MMP-8), interleukin-6 (IL-6), interleukin-1β (IL-1β), 8-hydroxydeoxyguanosine (8-OhdG), and Malondialdehyde (MDA).

As used herein, the term "marker" or "biomarker" refers to an indicator that is capable of reflecting a specific physical condition or state. As used herein, a "biomarker" may be a biological macromolecular substance (e.g., a protein, a nucleic acid), a small molecular substance (e.g., a compound), or any information or parameter of a biological sample (e.g., pH). Thus, as used herein, "information of a marker" or "value of a marker" refers to any information related to a marker. As one non-limiting example, if the biomarker is pH, then for a biological sample at pH 7.1, the value of the marker (e.g., information about the marker) may be 7.1.

As used herein, the term "sample" or "biological sample" or "specimen" refers to biological material isolated from a subject. The sample may comprise any biological material suitable for detecting the desired marker, and may comprise cellular material and/or non-cellular material from the subject. The sample may be isolated from any suitable biological tissue or fluid. In the present invention, the sample is gingival crevicular fluid. The sample may be pretreated prior to actual detection by filtration, distillation, extraction, concentration, dilution, inactivation of interfering components, addition of reagents, and the like.

As used herein, the terms "subject," "individual," and "patient" are used interchangeably to refer to a mammal from which a sample is taken, unless otherwise indicated. A subject, individual, or patient may have periodontitis, be at risk of developing periodontitis, or be suspected of having a predisposition to develop periodontitis. In some embodiments, a typical subject includes a human patient susceptible to, suffering from, or having suffered from one or more periodontitis. Thus, in some embodiments, the marker sets provided herein may be used to assess the stage of development (e.g., suffering from mild periodontitis or severe periodontitis) and/or the trend (e.g., worsening or improvement) of periodontitis in human patients already suffering from periodontitis.

As used herein, the term "predisposition, such as" predisposition to develop periodontitis, "refers to a reasonable medical probability of an event (e.g., occurrence or recurrence of periodontitis). The term "predisposition" also includes the frequency with which such events may occur before, after or during continued treatment.

The "reference value" of a marker may be an absolute or relative value or concentration of the marker, the presence or absence of the marker, a range of values or concentrations of the marker, a minimum and/or maximum value or concentration of the marker, an average value or concentration of the marker, and/or a median value or concentration of the marker. The reference value and the value of the marker of the subject may vary depending on the particular technique used to determine the marker in the biological sample.

The value of the marker of the subject can be compared to a reference value to which it is referenced using a variety of means, such as simple comparison, statistical analysis (e.g., t-test, wilcoxon rank sum test). In some embodiments, the above-described comparison may be accomplished by a manual and/or automated system (e.g., a test card, test instrument, provided below). In some embodiments, a subject's value of one or more markers that is 5%, 10%, 15%, 20% or more above the reference value to which it is referenced may be considered an increase in the value of the one or more markers of the subject.

"value" or "level" of one or more markers (or indicators) refers to the absolute (quantitative) or relative (qualitative) value or concentration of the biomarker in the sample.

Gingival crevicular fluid is fluid that permeates into the gingival crevicular from gingival connective tissue, and is composed mainly of serum, and other components are derived from adjacent periodontal tissues and bacteria. Gingival crevicular fluid from a subject may be collected and the amount of gingival crevicular fluid read using collection methods known in the art (e.g., capillary and filter paper strip methods). Preferably, the extraction device provided by the invention can be used for collecting gingival crevicular fluid samples from a subject.

The collected gingival crevicular fluid sample may be analyzed using a suitable method to detect the value of each marker in the biological sample.

Each label may be detected by photochemical, biochemical, spectroscopic, immunochemical, electrical, optical or chemical means. Exemplary suitable methods include immunochemical techniques such as chromatography (e.g., liquid Chromatography (LC), high Performance Liquid Chromatography (HPLC), gas Chromatography (GC)), mass spectrometry (e.g., mass Spectrometry (MS), tandem mass spectrometry (MS/MS 2)), combined methods (gas chromatography/mass spectrometry (GC-MS), liquid chromatography/mass spectrometry (LC-MS), ultra-high performance liquid chromatography/tandem mass spectrometry (UHLC/MS 2), and gas-chromatography/tandem mass spectrometry (GC/MS 2)), enzyme-linked immunosorbent assays (ELISA), and combinations thereof, refractive index spectroscopy (RI), ultraviolet spectroscopy (UV), fluorescent analysis, radiochemical analysis, near infrared spectroscopy (near IR), nuclear magnetic resonance spectroscopy (NMR), light scattering analysis (LS). Additionally, in some embodiments, the value of one or more markers may also be measured by way of indirect measurement, such as measuring the value of a compound (or compounds) that correlates to the value of the marker that is desired to be measured.

In some embodiments, preferred detection methods for some markers of the invention (e.g., gingival crevicular fluid hemoglobin, amounts of Porphyromonas gingivalis, matrix metalloproteinase-8, interleukin-6, interleukin-1 beta, 8-hydroxydeoxyguanosine, malondialdehyde, and gingival proteinase K) include immunoassays or aptamer-based assays. Such immunoassays or aptamer-based assays are conventional and well known in the art, involving the use of reagents (e.g., antibodies or aptamers) that are specific for the substance of interest. Reagents (e.g., antibodies or aptamers) that "specifically bind" to a substance of interest are terms well known in the art, and methods for determining specific binding are also well known in the art. In some embodiments, the invention detects the protein concentration of the label. Protein concentration is typically measured in mass per volume, e.g., mg/ml, μg/ml, and pg/ml.

In some embodiments, the pH of gingival crevicular fluid, a marker of the present invention, may be measured by a pH indicator method (e.g., methyl red and bromothymol blue), and may also be measured by a pH test paper (e.g., precision pH test paper) or a pH meter.

As used herein, a "detection reagent" is a reagent or compound that specifically (or selectively) binds, interacts with, or detects a label. In some embodiments, the detection reagent comprises an antibody, a polyclonal antibody, or a monoclonal antibody. In some embodiments, the detection reagent comprises a complex (e.g., an antibody-immobilized fluorescent particle) formed with an antibody, polyclonal antibody, or monoclonal antibody. In some embodiments, the detection reagent comprises an aptamer.

After determining the value of one or more markers in a biological sample from a subject, the value is compared to a reference value to which it is referenced. The reference values may include a fixed reference value for each marker and/or a reference value derived from itself. When a subject first evaluates the periodontitis development status by using the marker set provided by the present invention, the values of all markers in the marker set of the subject are usually compared with corresponding fixed reference values, so as to evaluate the periodontitis development status of the subject: if all of the marker values for the subject belong to the zero-phase reference value, the subject is assessed as periodontal health; if the value of part (or all) of the markers of the subject belongs to the first stage reference value and the other part (or none) belongs to the zeroth stage reference value, the subject is evaluated as a patient with mild periodontitis (or in the first stage of development of periodontitis); if the value of a portion (or all) of the markers of the subject belongs to the second stage reference value and another portion (or none) belongs to the zero or first stage reference value, the subject is assessed as a patient with severe periodontitis (or in a second stage of development of periodontitis). When the subject has at least once assessed its state of periodontitis by passing through the set of markers provided by the invention, the value of each marker of the subject may be compared to the last (or optionally one or more) detection result thereof to further assess the state of periodontitis of the subject.

Based on the results obtained above for the stages of periodontitis development, the subject's tendency to develop periodontitis (especially for subjects in the first and second stages of periodontitis) can be further assessed by comparing the value of all markers of the subject with a reference value derived from the subject. If the value of all markers in the subject's marker set is not significantly increased (i.e., not higher) than the reference value from which it is referenced (e.g., is the same as the reference value, is substantially the same as the reference value, is not statistically different from the reference value, is within a predetermined range of the reference value, etc.), then the subject is assessed as having a lower risk of exacerbation of periodontitis (which may be considered as a negative trend of not exacerbation of periodontitis) over a period of time since the biological sample was collected; if the value of one or more markers in the set of markers appears to be elevated compared to its reference value derived from itself, the subject is assessed as having a certain risk of exacerbation of periodontitis (i.e., a positive trend of exacerbation of periodontitis) over a period of time from the day the biological sample is collected. In particular, the value of all the markers of the marker set is increased compared to the reference value from which it is referenced, and then the subject is assessed as having a very high risk of exacerbation of periodontitis (i.e. a strong positive trend of exacerbation of periodontitis) over a period of time from the day the biological sample is taken. Of course, if the values of most or all of the markers in the set of markers are reduced (and the other markers are not significantly increased) compared to their reference values, then the subject is assessed as having a tendency to have an improvement in periodontitis over a period of time from the day the biological sample was collected.

The occurrence and development of periodontitis is not a continuous process, and has the development change in the acute phase and the possibility of recurrence after treatment. For patients with the same clinical symptoms, different degrees of periodontitis disease may be corresponded. The marker group provided by the invention can accurately and sensitively evaluate the periodontitis development state (such as the periodontitis development stage and the periodontitis development trend) of a subject from multiple dimensions, is beneficial to an oral doctor to evaluate, diagnose and monitor the periodontal health condition of a patient (i.e. clinical monitoring), is beneficial to the patient to simply and accurately self-evaluate, self-diagnose and self-monitor the periodontal health condition (i.e. patient self-monitor), avoids the extrusion of medical resources, and is especially suitable for patients suffering from periodontitis or being easy to relapse or having the risk of periodontitis deterioration. The marker group provided by the invention can evaluate the periodontitis development state change of a subject early (for example, a patient with mild periodontitis or a periodontal health crowd at risk of periodontitis deterioration can be accurately identified) and give an early warning (for example, oral care measures are required to be timely taken, oral hygiene habits are changed, OTC drugs are taken, and the like), so that the periodontitis of the crowd is prevented from being deteriorated to a certain extent. Of course, patients with severe periodontitis or patients with mild periodontitis and at risk of exacerbation of periodontitis, which are evaluated by the marker set according to the invention, need immediate hospitalization to prevent further exacerbation.

Because the marker set provided by the invention can evaluate the periodontitis state of the subject from multiple dimensions, an stomatologist or the subject himself can select a more targeted treatment scheme according to the actual detection result (for example, if the value of the porphyromonas gingivalis related marker in the detection result is obviously increased, antibacterial treatment aiming at the porphyromonas gingivalis can be selected), or in the case of abnormal porphyromonas gingivalis, macro-genome sequencing or 16sRNA sequencing can be further carried out, so that early monitoring of the oral cavity tooth Zhou Junqun disorder is facilitated.

In addition, the marker set provided by the invention can be used as one of the projects of periodic dental examination or part of large-scale screening so as to monitor the early diagnosis and the development state of periodontitis of subjects of a wider population periodically and dynamically, and is beneficial to early discovery and early treatment of periodontitis.

Further, the marker sets provided by the present invention are also useful for assessing the effect of a patient's treatment regimen. For patients diagnosed with periodontitis and treated over a period of time, the values of the sets of markers can be used to check whether periodontitis has been ameliorated (e.g., the value of one or more markers has decreased after a period of time treatment) and whether the treatment regimen needs to be adjusted (e.g., if the value of one or more markers has not increased significantly or has increased after a period of time treatment, indicating that the treatment regimen may not be appropriate for the subject), thus facilitating the implementation of personalized treatment.

In particular, the marker sets provided by the invention are suitable for dynamic monitoring of periodontitis, for example, authorized stomatologists and subjects can obtain historical data of each marker obtained by each detection on the terminal.

Example 1

This example incorporates 18 patients with mild periodontitis, 16 patients with severe periodontitis, and 22 periodontal healthy populations. Wherein, the clinical diagnosis standard of periodontitis is: there is a loss of interproximal adhesion (CAL) for 2 or more non-adjacent teeth, or a loss of adhesion of 3mm to the buccal (lingual) surface of > 2 teeth, with a periodontal pocket > 3 mm. Clinical diagnostic criteria for mild periodontitis (including mild periodontitis and moderate periodontitis) are: the gum has inflammation and Bleeding (BOP), the tooth Zhou Dai mm and adhesion loss is 1-2 mm, and the X-ray film shows that the alveolar bone absorption is not more than 1/3 of root length. Clinical diagnostic criteria for severe periodontitis were: the periodontal pocket is more than 6mm, the adhesion loss is more than or equal to 5mm, the X-ray film shows that the alveolar bone absorption exceeds 1/2 of root length, a plurality of teeth have root bifurcation lesions, the teeth have loosening, and the inflammation is more obvious or periodontal abscess can be caused. Diagnostic criteria for clinically healthy periodontal disease are: normal periodontal tissue has no or very little clinical inflammation, the depth of periodontal probing is no more than 3mm, and the site of probing bleeding is less than 10%. All subjects had no severe systemic disease or immune system disease.

The invention finds that the accurate distinction between periodontal healthy people and patients with mild periodontitis can be achieved by combining five dimensional markers, namely the amount of gingival crevicular fluid, the pH of gingival crevicular fluid, gingival crevicular fluid hemoglobin, a porphyromonas gingivalis related marker and a periodontitis related biochemical marker, as shown in Table 1.

TABLE 1 differentiation between periodontal health population and patients with mild periodontitis

The accuracy related by the invention is calculated by the following formula:

where TP represents True Positive (True Positive), TN represents True Negative (True Negative), FP represents False Positive (False Positive), and FN represents False Negative (False Negative).

Overall, the combination of the five-dimension markers is adopted as a marker group for detecting periodontitis, so that the distinction between periodontal healthy people and mild periodontitis patients can be realized more specifically, comprehensively, accurately and effectively. This helps in early diagnosis of periodontitis, and even enables diagnosis of periodontitis earlier than the current clinical examination.

The above data also shows that for areas of underdeveloped economics or conditions where accuracy requirements are relatively low (e.g., large-scale screening), the three dimensions of gingival crevicular fluid, pH of gingival crevicular fluid, gingival crevicular fluid hemoglobin, or the four-dimensional set of markers of gingival crevicular fluid, pH of gingival crevicular fluid, gingival crevicular fluid hemoglobin, and periodontitis-associated bacteria can be selectively employed, while ensuring a degree of accuracy, the number of markers detected can be reduced to reduce the cost of the detection scheme and promote the popularity of periodontitis detection.

Example two

Next, this example further verifies whether the above marker set can accurately identify patients with mild periodontitis and patients with severe periodontitis, and the results are shown in table 2. The result shows that the marker group provided by the invention not only can realize the distinction between periodontal healthy people and patients with mild periodontitis, but also can realize the accurate diagnosis of patients with mild periodontitis and patients with severe periodontitis. In a word, the marker set provided by the invention can accurately identify and distinguish the development stage of periodontitis and can be used as an effective basis for periodontitis detection.

TABLE 2

In clinical examinations, the limits of early development and periodontal health of periodontitis are not very obvious, which not only gives the dentist trouble, but also prevents "early discovery, early treatment" of periodontitis. As shown in table 3, the marker set provided by the present invention not only can accurately identify and distinguish the development stage of periodontitis (zeroth stage-periodontal health, first stage-mild periodontitis, and second stage-severe periodontitis), but also can judge the development trend of periodontitis (e.g., worsening or improvement of periodontitis) of a subject according to the value of each marker in the marker set measured by the subject each time. This not only provides early warning of deterioration of periodontitis but also provides a new angle for evaluating treatment regimens, e.g., whether the subject's stage of periodontitis progression transitions from the first stage or the second stage to the zeroth stage, as determined by the value of each marker, to evaluate whether the subject's periodontitis is alleviated, controlled, ameliorated, or restored to a healthy state by the accepted periodontal treatment.

The marker group provided by the invention can evaluate the periodontitis state of a subject from multiple dimensions, and the determination result is easy to visualize (for example, the determination result can be further processed to more intuitively reflect the determination result), so that the method is beneficial to an stomatologist to "dose symptomatically" according to the actual determination result of the subject, personalized treatment is better realized, and the accurate evaluation of a treatment scheme and the maintenance condition after treatment by the stomatologist and/or periodontitis patients is also beneficial.

TABLE 3 Table 3

Example III

In order to facilitate early evaluation of the change in the development status of periodontitis in a subject, or to rapidly screen out subjects who may have periodontitis or subjects at risk of deterioration of periodontitis from a large number of subjects in oral examination activities of a large population of subjects, the present invention provides an extraction device for extracting a gingival crevicular fluid sample, as shown in fig. 1 to 5, comprising:

an extraction tube 10, said extraction tube 10 comprising an extraction end 12 and an outlet end disposed on a side remote from said extraction end, wherein said extraction end 12 is sized and shaped to extend between teeth and adjacent gums, said extraction tube 10 being provided with a capillary channel 11 on a side proximate to said extraction end 12 and at least one air pressure equalization tube 13 disposed thereon;

A mixing chamber 20, the mixing chamber 20 comprising a receiving space 21 for receiving the sample, the receiving space 21 being in communication with the outlet end, and the receiving space being provided with a first opening at a location corresponding to the outlet end;

a pushing part 30, a first end 31 of which can extend into the extraction tube 10 from the outlet end, and a second end 32 of which penetrates the accommodating space 21 and extends out from a first opening provided in the accommodating space 21; wherein, when the pushing part 30 moves along a first direction, the action of the pushing part on the air inside the extraction tube can drive the sample in the capillary channel 11 to flow along the first direction, wherein the first direction is the direction from the extraction end to the outlet end.

In some embodiments, the extraction tubes 10 are all provided as elongate capillary channels.

In some embodiments, the side of the extraction tube 10 that is on the extraction end is provided as a capillary channel, while the side thereof that is near the outlet end may be provided as a channel of conventional size (i.e., a non-capillary channel).

In the embodiment of the invention, the extraction tube can spontaneously collect gingival crevicular fluid by utilizing capillary action, so that the extraction amount of the obtained gingival crevicular fluid is related to the degree of periodontitis of a subject to a certain extent. For example, a subject with severe periodontitis may extract more gingival crevicular fluid at the same level of capillary action and the same extraction time.

In some embodiments, the extraction tube 10 is provided with at least one length of graduation 14 that can be used to read the amount of extraction of the sample.

In some embodiments, the capillary channel has an inner diameter of about 0.4mm to about 0.6mm.

In some embodiments, two sections of scale marks are sequentially arranged on the capillary channel at intervals, and the readings of gingival crevicular fluid contents corresponding to the two sections of scale marks are respectively: 10. Mu.L, 20. Mu.L.

Or, in some embodiments, four sections of scale marks are sequentially arranged on the capillary channel at intervals, and the readings of gingival crevicular fluid contents corresponding to the four sections of scale marks are respectively: 10. Mu.L, 20. Mu.L, 40. Mu.L, 50. Mu.L.

For example, in some embodiments, the capillary channels have an inner diameter of about 0.4mm, and the length of the corresponding capillary channel is set to about 10cm-12cm. And a section of scale marks are respectively arranged at the positions of about 2cm, 4cm, 8cm and 10cm away from the extraction end on the capillary channel. Wherein, when the liquid level of gingival crevicular fluid is between scale marks of 2cm-4cm, the content of gingival crevicular fluid is about 10-20 mu L; when the level of gingival crevicular fluid is between the scale marks of 4cm-8cm, it means that the gingival crevicular fluid is present in an amount of about 21-40 μl; when the level of gingival crevicular fluid is between 8 and 10cm, this means that the gingival crevicular fluid is present in an amount of between about 41 and 50. Mu.L. In this embodiment, the content interval of gingival crevicular fluid can be rapidly read by means of scale marks.

For another example, in some embodiments, the capillary channel has an inner diameter of about 0.5mm and a length corresponding to about 7-10cm. And a section of scale marks are respectively arranged on the capillary channel at positions which are about 1.3cm, 2.5cm, 5.1cm and 6.4cm away from the extraction end. Wherein, when the liquid level of gingival crevicular fluid is between 1.3cm and 2.5cm of scale marks, the content of gingival crevicular fluid is about 10-20 mu L; when the level of gingival crevicular fluid is between the scale marks of 2.5cm to 5.1cm, this means that the gingival crevicular fluid is present in an amount of about 21 to 40 μl; when the level of gingival crevicular fluid is between 5.1 and 6.4cm, this means that the gingival crevicular fluid content is between about 41 and 50. Mu.L.

For another example, in other embodiments, the capillary channel has an inner diameter of about 0.6cm and a length corresponding to about 4.5 cm to about 6cm. Correspondingly, a section of scale marks are respectively arranged on the capillary channel along the positions 0.9cm, 1.8cm, 3.5cm and 4.4cm away from the extraction end.

In the embodiment of the invention, the spontaneous collection effect of the extraction tube and the arrangement of the scale marks are convenient for a user to accurately and rapidly quantitatively read the extraction amount of gingival crevicular fluid. It will be appreciated that the gingival crevicular fluid extraction reading may be recorded as a specific value (and correspondingly, more graduations may be provided on the tube for more accurate reading) or as a range of values during use.

In some embodiments, the extraction end is rounded in order to avoid or reduce damage to gums during extraction.

In some embodiments, the first end 31 of the pushing portion has an outer diameter dimension that is greater than an outer diameter dimension of the second end 32, and correspondingly, the outlet end of the extraction tube has an inner diameter dimension that is greater than an inner diameter dimension of the first opening.

For example, in some embodiments, as shown in fig. 2 and 3, the outlet end of the extraction tube 10 extends into the accommodating space 21, and a first opening through which the pushing portion 30 passes is further provided on the accommodating space 21 (specifically, the first opening may be a tubular structure formed by extending the accommodating space outwards, and the first opening is located on an extension line where the center line of the extraction tube is located). The inner diameter of the outlet end is slightly larger than that of the first opening, and the first end 31 of the pushing portion is larger than that of the second end 32 of the pushing portion, so that the first end of the pushing portion can reciprocate in the extraction tube 10, but the first end of the pushing portion cannot pass through the first opening, that is, the pushing portion cannot be pulled out completely, so that the moving distance of the pushing portion is limited to a certain extent.

For another example, in some embodiments, a limiting ring is disposed on a side of the pushing portion near the first end, and an outer diameter of the limiting ring is larger than an inner diameter of the first opening, so as to limit a moving distance of the pushing portion, and avoid the pushing portion from being pulled out completely.

In some embodiments, the pushing portion is made of a rubber material, and an outer diameter dimension of the pushing portion in a natural form (i.e., a state in which no external force acts) is larger than an inner diameter dimension of the first opening. Therefore, when the pushing part made of the rubber material is in contact with the outlet end and the first opening, the rubber material is extruded and deformed to a certain extent under the extrusion action of the first opening and the outlet end, so that the pushing part is in close contact with the first opening and the outlet end, and a good sealing effect is achieved. And the rubber material is tightly contacted with the outlet end and the first opening, so that the pushing part can be directly pulled out completely (which is equivalent to providing a first resistance for limiting the movement of the pushing part) due to overlarge external acting force in the use process can be avoided to a certain extent.

In some embodiments, the extraction tube is configured as a frangible structure proximate a side of the mixing chamber. After extraction is completed using the extraction device, the section of tubing between the break-off structure and the extraction end may be broken off to primarily use the mixing chamber 20 as the final sample storage unit.

Alternatively, in other embodiments, the extraction tube 10 comprises a fixed tube and a movable tube that are removably connected to each other.

Specifically, in some embodiments, the fixed tube is fixedly connected to the mixing chamber, and one end of the fixed tube extending into the accommodating space is an outlet end, and the other end of the fixed tube is detachably connected to one end of the movable tube. The other end of the movable tube is the extraction end, and the movable tube is arranged into a capillary structure. In this embodiment, the movable tube that is in direct contact with the oral cavity of the subject is required to be used as a disposable, and other parts of the extraction device can be reused after cleaning and disinfection, so that the cost for detecting periodontitis can be reduced to a certain extent.

Further, in some embodiments, the stationary tube is made of a rubber material (or, in other embodiments, the portion of the stationary tube that mates with the movable tube is made of a rubber material), the movable tube is made of a plastic, and the outer diameter dimension of the movable tube may be slightly larger than the inner diameter dimension of the rubber material. Wherein the movable tube is connected with the fixed tube by being inserted into the inner wall of the fixed tube. In this embodiment, the fixed pipe and the movable pipe can be tightly matched (avoid liquid overflow), and meanwhile, the disassembly and replacement between the fixed pipe and the movable pipe are simple and easy to operate.

In some embodiments, the outlet end of the extraction end may also be used as a liquid outlet of the receiving space.

Alternatively, in other embodiments, the receiving space is provided with a liquid outlet and the liquid outlet is provided with a sealed opening. In the preservation and transportation process of the extraction device, the sealing opening can ensure the cleaning of the inner space. The sealing opening may be opened when it is desired to take out the sample from the receiving space.

In some embodiments, the mixing chamber 20 has a pre-stored diluent therein. Suitable diluents are, for example, TES, MES, PIPES, HEPES, MOPS, tris, PBS and physiological saline.

In some embodiments, the air pressure balance tube 13 is provided with a sealed opening, and when the gingival crevicular fluid is extracted by using the extracting device, the air pressure balance tube 13 is required to balance the air pressure inside and outside the capillary channel, and then the sealed opening is opened. When it is necessary to pull the pushing part 30 so that the liquid inside the capillary channel can enter the accommodating space 21, the seal opening may be closed to limit the action of the air pressure balance tube 13 to balance the air pressure.

Alternatively, in other embodiments, the air pressure equalization tube 13 is made of a pliable material (e.g., plastic), wherein the air pressure equalization tube extends a length outwardly from the main body portion of the extraction tube. When the action of the air pressure balance tube to balance the air pressure is required to be limited, the bent air pressure balance tube can be pressed (even if the air pressure balance tube achieves a sealing effect, air exchange or flow between the inside of the extraction tube and the external environment cannot be assisted any more) so that the liquid sample inside the capillary channel can smoothly enter the accommodating space 21 under the action of the pushing part.

In some embodiments, a label is also provided on the extraction device (e.g., on the outer surface of the mixing chamber). After the user reads the extraction amount of gingival crevicular fluid, the extraction amount reading can be recorded on the label, so that the data recording management is facilitated.

In order to further explain the technical solution in the embodiment of the present invention, the following describes the use process of the extraction device provided in the foregoing with reference to fig. 5:

when the extraction device is not activated, the first end of the pushing part of the extraction device is positioned at the inner side of the outlet end of the extraction tube so as to seal the diluent pre-stored in the accommodating space (prevent the diluent from entering the extraction tube).

In use, as shown in fig. 5, the extraction end of the extraction device is extended between the tooth and the adjacent gums, such that gingival crevicular fluid wicks into the interior of the capillary channel. After the extraction is finished, the extraction amount of gingival crevicular fluid is read through the scale marks, and corresponding data is recorded. The gingival crevicular fluid extraction device is then inverted (or placed in a horizontal orientation) and the push portion is pulled in a first direction such that fluid within the capillary channel enters the mixing chamber under the dual action of gravity and the push portion and mixes with the pre-stored diluent.

Finally, the lower half of the extraction tube 10 (i.e., the side near the extraction end) is broken off from the breaking structure 15, and the pushing part 30 is pushed back again to the outlet end of the extraction tube 10, so that the mixed solution of gingival crevicular fluid and diluent in the mixing chamber 20 is hermetically preserved.

In this embodiment, the user (e.g., a inspector) can conveniently and rapidly extract the gingival crevicular fluid and mix the gingival crevicular fluid with the diluent. The gingival crevicular fluid and the diluent are mixed and stored, so that loss of the gingival crevicular fluid can be reduced to a certain extent, and storage time of the gingival crevicular fluid is prolonged. In addition, since the mixed preservation mode of gingival crevicular fluid and diluent is adopted in the embodiment, loss of gingival crevicular fluid in the processes of transportation, storage and the like can be reduced, and conversely, necessary extraction amount of gingival crevicular fluid (namely, gingival crevicular fluid amount capable of meeting detection requirements) can be reduced to a certain extent. Therefore, the number of extraction operations or the extraction operation time of gingival crevicular fluid can be reduced in the actual extraction process, so that the damage of the extraction device to the gums of a subject (such as periodontitis patients) can be reduced or avoided to a certain extent, and the extraction efficiency of gingival crevicular fluid can be improved to a certain extent.

Meanwhile, the mixing cavity 20 with a more regular structure and smaller occupied space is used as an independent storage unit, so that the whole storage and transportation of the extraction device (or gingival crevicular fluid extracted in batches) after batch use are facilitated.

Under the synergistic effect of the multiple effects of smaller gum injury, higher extraction efficiency, convenience in batch storage and the like, the gingival crevicular fluid extraction device disclosed by the invention can be well applied to the application scene of rapidly screening the oral health condition (such as whether periodontitis exists) of a large-scale crowd.

For example, the extraction device of the invention can be widely applied to the activities of oral cavity consultation activities held by organizations such as communities aiming at local residents, or activities of free oral health detection of teachers and students in healthy oral cavity campuses and the like. In these large-scale screening activities, the number of subjects is large, and the sampling and detection work is very heavy and the efficiency requirement is very high. And such large-scale screening activities have also involved long-term on-site sampling, off-site detection, etc. (e.g., requiring residents to be sampled on-site for a long period of time in an area remote from the hospital or detection laboratory, and then transported to the hospital or detection laboratory). On the other hand, the extraction device provided by the invention can reduce the necessary extraction amount of gingival crevicular fluid, so that the sampling efficiency is greatly improved when the device is used for sampling a single subject, and the high-efficiency sampling can effectively reduce the gum damage to the subject. After the sampling is finished, a user can also quickly record the extraction amount reading of the gingival crevicular fluid and quickly realize the mixed storage of the gingival crevicular fluid and the diluent.

The batch extraction device used for many subjects can be stored and transported in batch in a minimized unit form (i.e. a mixing chamber) so as to facilitate convenient transportation of the sampled products from sampling sites such as communities, campuses and the like to hospitals or detection laboratories in different places to complete subsequent detection. In addition, during the storage and transportation process, the mixed storage can reduce the loss of gingival crevicular fluid so as to prolong the effective preservation time of the gingival crevicular fluid.

Preferably, for such large-scale batch test items, the amount of gingival crevicular fluid may be determined by a); b) The pH of gingival crevicular fluid; c) The periodontal disease patients and periodontal disease patients can be rapidly distinguished by three dimensional markers such as gingival crevicular fluid hemoglobin, so that early discovery and early treatment of periodontitis can be realized through lower economic cost.

Example IV

As shown in fig. 6 to 9, the present embodiment provides another extraction device for extracting a gingival crevicular fluid sample, the extraction device including:

a first storage chamber 40, wherein a first storage space 42 for accommodating the sample is formed in the first storage chamber 40, the first storage chamber 40 further comprises a pushing part 30, a first end 31 of the pushing part 30 is positioned inside the first storage chamber 40, and a second end 32 of the pushing part 30 extends to the outside of the first storage chamber 40;

An extraction tube 10, said extraction tube 10 comprising an extraction end 12 and an outlet end disposed on a side remote from said extraction end 12, wherein said outlet end is in communication with said first storage space 42, said extraction end 12 is sized and shaped to extend between a tooth and an adjacent gum, and a side of said extraction tube 10 adjacent said extraction end 12 is provided as a capillary channel 11, and said outlet end of said extraction tube 10 is in communication with said second storage space;

a second storage chamber 50, wherein a second storage space 52 for containing liquid is formed in the second storage chamber 50, and the second storage space is communicated with the first storage space through a first channel, at least one air pressure balancing port is further arranged in the second storage space 52, the second storage chamber 50 is outwardly extended with a first channel 53, the first channel 53 is communicated with the second storage space 52, and one side of the first channel 53 close to the second storage space is provided with a capillary structure;

when the pushing portion 30 moves in a direction away from the first storage cavity 40, the action of the pushing portion 30 on the air inside the extraction device can drive the sample in the capillary channel and the liquid in the second storage cavity 50 to flow in a direction approaching to the first storage cavity 40.

It should be noted that in the embodiments of the present invention, different terms, such as "accommodating space", "first storage space" or "second storage space", are used to describe a chamber for storing a liquid sample (such as gingival crevicular fluid or a diluted fluid), such as a mixing chamber, a storage chamber, or the like, which is mainly used to facilitate distinguishing between different chambers in different embodiments. In fact, in the embodiment of the present invention, the above various accommodating spaces or storage spaces may be of the same structure. Of course, different configurations are also possible. The structure in which these chambers may be provided includes: spherical cavities, or rectangular cavities, etc.

In some embodiments, the extraction tube 10 includes a fixed tube fixedly connected to the second storage space 52, and a movable tube having one end detachably connected to the fixed tube, and the other end of the movable tube is the extraction end.

In this embodiment, the movable tube portion is a disposable portion, and the remaining portion of the extraction device can be reused a plurality of times after corresponding cleaning operations such as sterilization. The cost of periodontitis detection is reduced to a certain extent.

For example, in some embodiments, the stationary tube is made of a rubber material (or, in other embodiments, the portion of the stationary tube that mates with the movable tube is made of a rubber material) and the movable tube is made of plastic, wherein the movable tube is inserted into the stationary tube inner wall to connect with the stationary tube. In this embodiment, the fixed pipe and the movable pipe can be tightly matched (avoid liquid overflow), and meanwhile, the disassembly and replacement between the fixed pipe and the movable pipe are simple and easy to operate.

In some embodiments, the first storage space 42 is provided with a second opening 41.

In this embodiment, the second opening 41 may function to balance the air pressure. When the gingival crevicular fluid is extracted from the gingival of the subject using the extraction tube, the second opening is left open so as to be usable for balancing the air pressure inside the extraction device.

Further, in some embodiments, the second opening may also function as a fluid outlet, i.e. for draining a mixture of gingival crevicular fluid, diluent fluid inside the second storage cavity.

In some embodiments, a sealed opening is provided over the second opening. During the preservation and transportation of the extraction device, the sealing opening can seal the second opening to ensure the cleaning of the inner space. The sealed opening may be opened when the second opening is required to perform the function of draining liquid or balancing air pressure.

In some embodiments, the extraction tube is provided with at least one section of graduation marks useful for reading the amount of extraction of the sample.

In some embodiments, the extraction end is rounded.

In some embodiments, the first end of the pushing portion has a dimension that is greater than a dimension of the second end.

In some embodiments, the pushing portion is made of a rubber material.

In some embodiments, the first channel is a capillary channel for connecting the first and second storage spaces, and the capillary channel may act as a flow restrictor for the diluent in the second storage space.

Specifically, the first end of the capillary channel is connected to the first storage space, and the second end is connected to the second storage space. At this time, since the inner diameter of the first end of the capillary channel is much smaller than the size of the second storage space, there may be a portion of the diluent in the second storage space that enters the first end of the capillary channel under the capillary action (it will be understood that the diluent content entering the capillary channel under the action of the air pressure will not be excessive since the second storage space is in a sealed state). Also, since the size of the first storage space (specifically, the inner diameter of the junction of the first storage space and the capillary channel) is larger than the inner diameter of the second end of the capillary channel, it is difficult for the liquid entering the capillary channel to continue into the first storage space through the second end (or, the size of the first storage space is larger than the size of the second end of the capillary channel), which creates a second resistance to the diluent that limits its flow to the first storage space. Therefore, the first channel in this embodiment has a flow limiting effect, so that the diluent is further prevented from entering the extraction tube and contaminating the extraction tube.

Of course, in other embodiments, the first channel may be formed by a combination of a common sized channel and a capillary channel. Specifically, the side of the first channel near the second storage space is provided as a capillary channel.

In some embodiments, to avoid the elongated first channel from being broken, a protective sleeve is also provided outside the first channel.

It can be understood that, unlike the third embodiment of the present invention, in this embodiment, a relatively independent storage space is designed for the diluent, and a corresponding flow limiting structure (i.e., the first channel) is adopted to assist in achieving stable storage of the diluent. The storage amount of the diluent can be increased to a certain extent, and the diluent cannot be immersed into the extraction tube and pollute the extraction tube because of excessive diluent. Therefore, compared with the extraction device in the third embodiment, the extraction device in the present embodiment can provide more detection samples (i.e. the mixed solution of gingival crevicular fluid and diluent) to a certain extent, so as to satisfy the detection scenario with relatively more detection items.

For example, for oral patients who are self-visiting, re-diagnosing to a hospital, their propensity to develop periodontitis or the severity of periodontitis can be relatively high. Therefore, for such patients who are subjected to autonomous diagnosis, providing sufficient detection samples facilitates the selection of a more comprehensive and accurate detection scheme. Specifically, the patient for autonomous diagnosis can comprehensively detect the quantity of gingival crevicular fluid, the pH value of gingival crevicular fluid, gingival crevicular fluid hemoglobin, the quantity of porphyromonas gingivalis, and a plurality of indexes such as gingival proteinase K, matrix metalloproteinase-8, interleukin-6, interleukin-1 beta, 8-hydroxydeoxyguanosine, malondialdehyde and the like.

It can be understood that the above is only to illustrate a preferred application scenario of the embodiment of the present invention, and in fact, the extraction device in both the third embodiment and the fourth embodiment may be applied to different screening activities such as large-scale community detection or field detection in hospitals.

The following further describes the technical scheme in the embodiment of the present invention in combination with the use process of the extraction device:

as shown in fig. 9, the extraction end of the extraction device extends into the gums of the subject to extract gingival crevicular fluid, and the second opening is then required to be opened so that the second opening acts to balance the air pressure.

After the gingival crevicular fluid is extracted, the second opening is closed, the extraction device is inverted, and the pushing part is pulled outwards along the direction away from the first storage space, so that the diluent in the second storage space enters the first storage space under the action of gravity and the action of the pushing part (in the process, the second resistance is overcome); likewise, gingival crevicular fluid in the capillary channel enters the first storage space under the force of gravity and under the force of the pushing portion. Subsequently, gingival crevicular fluid and diluent fluid are mixed with each other in the first storage space. Finally, the mixed liquid can be discharged by utilizing the second opening so as to complete the corresponding detection project.

Example five

As shown in fig. 10a to 10c, the present embodiment further provides a detection card matched with the extracting device of any one of the third and fourth embodiments, where the detection card includes:

a body 60;

a sample inlet 61 provided on the body, an opening of the sample inlet being provided on the first surface of the body;

the first detection areas 62 are connected to the sample loading port 61 through a second channel 63, wherein a first end of the second channel is connected to the sample loading port 61, and a second end of the second channel is connected to the first detection areas 62.

In some embodiments, a plurality of first detection zones 62 may be provided. For example, it may be generally set to 4 to 9.

In some embodiments, a reagent may be pre-stored within the first detection zone, which may be used to detect the value of the label or the value of a compound associated with the value of the label, wherein the reagent may be an aptamer corresponding to the label (corresponding to the target). For example, in one embodiment, 9 first detection areas are provided on the detection card, wherein 1 first detection area is used as a blank, the other 8 first detection areas are used as test sets, and the 8 first detection areas are respectively provided with an aptamer specifically recognizing human hemoglobin, an aptamer specifically recognizing Porphyromonas gingivalis, an aptamer specifically recognizing matrix metalloproteinase-8, an aptamer specifically recognizing interleukin-6, an aptamer specifically recognizing interleukin-1 beta, an aptamer specifically recognizing 8-hydroxydeoxyguanosine, an aptamer specifically recognizing malondialdehyde, and an aptamer specifically recognizing gingival proteinase K.

Of course, any one or more of the above reagents may be pre-stored on the test card according to the current actual test requirements (e.g., test accuracy).

In some embodiments, the aptamer is a single-stranded DNA molecule.

In some embodiments, the aptamer is modified with a fluorescent group and/or a quenching group. For example, fluorescent labeling techniques can attach a covalent bond of a fluorophore to an aptamer. This technique provides a signal of the object to be detected by reacting a specific group of a fluorescent signal substance with a specific group of an aptamer, thereby completing the labeling process thereof, and utilizing the fluorescent characteristic of the fluorescent group. Among them, rhodamine and fluorescein are most commonly used as the reagent for providing fluorescent genes, and other fluorescent labeling reagents such as polycyclic aromatic hydrocarbon compounds, aromatic heterocyclic compounds, chelates of rare earth elements and the like can be selected.

In some embodiments, the aptamer may be further prepared into an aptamer sensor using a nanomaterial as a carrier, where the nanomaterial is selected from the following materials: the metal and metal oxide nanoparticles include gold nanoparticles, silver nanoparticles (silver nanoparticles, agNPs), platinum nanoparticles (platinum nanoparticles, ptNPs), iron oxide nanoparticles (iron oxide nanoparticles, IONPs), molybdenum nanoparticles, and the like.

In some embodiments, a second detection area is further provided on the detection card, and the second detection area is used for detecting the PH value of the mixed solution. For example, the second detection area is a rectangular groove arranged on the surface of the detection card, and a PH test paper can be arranged in the rectangular groove for detecting the PH value of the mixed solution. In this embodiment, a user (e.g., a detecting person) may read the PH by manually comparing with the standard color chart.

In some embodiments, the second channel 63 may be disposed inside the test card.

Alternatively, in other embodiments, the second channel (i.e., the sample separation channel) may be disposed on the second surface of the test card and sealed by covering the second surface with a sealing film.

In some embodiments, the second end of the second channel 63 is configured as a capillary channel, and the inner diameter of the capillary channel is configured to decrease gradually in a direction approaching the first detection zone 62.

In this embodiment, when the reagent in the liquid state is stored in the first detection region 62, the inner diameter gradient of the second channel 63 can limit the reagent in the first detection region 62 from overflowing into the second channel 63. Alternatively, even if some reagent overflows into the second end of the second channel 63, the overflow reagent in the second end is prevented from moving further toward the first end of the second channel 63 and even into the sample application port 61, and contaminating the test card (it will be appreciated that liquid will not actively flow from a location where capillary action is greater to a location where capillary action is less when no other external force is applied). Conversely, when the mixed solution in the sample adding port enters the first detection area 62 along the second channel 63, the second channel can also play a role in guiding and assisting the flow of the mixed solution, and the sample separating process of the mixed solution is not adversely affected.

In some embodiments, the second channel may be configured to have some capillary action.

For example, in one embodiment, in use, the mixed fluid is added to the sample application port via the fluid outlet of the gingival crevicular fluid extraction device, and the mixed fluid may be wicked (or may be combined with centrifugal force) through the second channel and into each of the first detection zones. The test card is then inserted into the test instrument in a horizontal arrangement (in other embodiments, it may also be in a vertical arrangement) and the test instrument may perform a data reading (e.g., reading the color of the test area) of each of the first test areas, wherein the use of the test card with the test instrument 70 may be seen in fig. 11-13.

Of course, it will be appreciated that the test card may also be used in other ways than shown in FIGS. 11-13 to interact with the test instrument.

Specifically, in some embodiments, the test instrument includes a housing having a test card placement cavity disposed therein for placing a test card, and a first test assembly comprising: the first image acquisition module (such as a fluorescence microscope or other fluorescence cameras used for acquiring fluorescence pictures of the first detection area) is arranged corresponding to the detection card placement cavity, and the first image analysis module is connected with the first image acquisition module. Wherein, after the user places the detection card in the detection card placement cavity and starts the detection instrument, the main workflow of the detection instrument is as follows: 1) The fluorescence microscope respectively acquires fluorescence images of each first detection area and sends the acquired images to the first image analysis module; 2) The first Image analysis module (for example, image J Image analysis software) obtains an Image, splits the Image into 3 channels of red, green and blue, and then performs fluorescent signal identification on the green channel map to obtain average fluorescent intensity.

Wherein the average fluorescence intensity (corresponding to the value of the label) in each first detection zone can be compared with the fluorescence intensity information (corresponding to the reference value of the "label") of different standard samples. For example, taking hemoglobin detection as an example, multiple sets of hemoglobin standard samples of different concentrations may be prepared in advance prior to detection, and fluorescence intensity information (e.g., average fluorescence intensity) after these different concentrations of hemoglobin standard samples have reacted with the same reagent (e.g., an aptamer that specifically recognizes human hemoglobin) in the same reaction environment may be tested and recorded. Finally, comparing the fluorescence intensity of the test sample with the fluorescence intensity of the standard sample by a detection personnel or corresponding computer software to obtain a corresponding judgment result (for example, the hemoglobin concentration of the sample).

For another example, in some embodiments, the detection instrument further comprises: a second detection assembly, wherein the second detection assembly comprises: the second image acquisition module is arranged towards a second detection area on the detection card, and the second image analysis module is connected with the second image acquisition module. Specifically, the second image acquisition module is a video camera (or also a camera), and the second image analysis module is a single-chip microcomputer (or also other electronic chips). After the first detection assembly completes detection work, the second detection assembly starts to operate, wherein the camera shoots an image of the second detection area and sends the image to the singlechip, and the singlechip measures the PH value of a sample to be detected in the second detection area through the contrasting color block.

Preferably, the second detecting assembly may further include: and an illumination module, which is likewise disposed toward the second detection zone, and which can provide an appropriate light source when the camera is taking an image.

Therefore, under the mutual cooperation of the detection card and the detection instrument, the collected gingival crevicular fluid can be subjected to detection reactions with various different dimensions at one time, and the detection instrument can synchronously output the values of various markers or the comparison result of the values of the markers and the reference value, so that the periodontitis development state of a subject can be conveniently and rapidly estimated.

Example six

The present embodiment provides a periodontitis detection kit, which includes the extraction device in the third embodiment or the fourth embodiment.

Further, in some embodiments, the periodontitis detection kit further comprises: any of the test cards of the fifth embodiment.

The embodiments of the present invention have been described above with reference to the accompanying drawings, but the present invention is not limited to the above-described embodiments, which are merely illustrative and not restrictive, and many forms may be made by those having ordinary skill in the art without departing from the spirit of the present invention and the scope of the claims, which are to be protected by the present invention.

Claims

1. A periodontitis detection kit, comprising:

-an extraction tube (10) comprising an extraction end (12), and an outlet end arranged at a side remote from the extraction end (12), wherein the extraction end (12) is sized and shaped to extend between a tooth and an adjacent gum, wherein the side of the extraction tube adjacent to the extraction end (12) is provided with a capillary channel (11), and wherein the extraction tube is provided with at least one air pressure balancing tube (13);

-a mixing chamber (20), the mixing chamber (20) comprising a receiving space (21) for receiving the sample, the receiving space (21) being in communication with the outlet end, and the receiving space being provided with a first opening at a position corresponding to the outlet end; the extraction pipe (10) comprises a fixed pipe fixedly connected with the accommodating space and a movable pipe with one end detachably connected with the fixed pipe, and the other end of the movable pipe is the extraction end (12);

a pushing part (30), a first end (31) of the pushing part (30) can extend into the interior of the extraction tube from the outlet end, and a second end of the pushing part (30) penetrates the accommodating space and extends outwards from the first opening; wherein when the pushing part (30) moves along a first direction, the action of the pushing part (30) on the air in the extraction tube can drive the sample in the capillary channel (11) to flow along the first direction, wherein the first direction is the direction from the extraction end (12) to the outlet end;

When the extraction device is not started, the first end of the pushing part is positioned at the inner side of the outlet end of the extraction pipe so as to seal the diluent pre-stored in the accommodating space.

2. Kit according to claim 1, characterized in that the extraction tube (10) is provided with at least one section of graduation marks (14) for reading the extraction quantity of the sample;

and/or the extraction tube is provided with a frangible structure (15) at the side close to the mixing chamber (20);

and/or the extraction end (12) is rounded;

and/or the first end (31) has an outer diameter greater than the second end (32), and correspondingly the outlet end has an inner diameter greater than the first opening;

and/or the pushing part (30) is made of rubber material.

3. Kit according to claim 1, characterized in that the receiving space (21) is provided with a liquid outlet (22) and that the liquid outlet is provided with a sealing opening.

4. The kit of claim 1, further comprising:

a test card, and the test card comprises:

a body (60);

a sample addition port (61) provided on the body, an opening of the sample addition port being provided on a first surface of the body (60);

The first detection areas (62) are communicated with the sample adding port through a second channel (63).

5. The periodontitis detection kit of claim 4, wherein the first detection zone is pre-stored with a corresponding reagent comprising: one or more of an aptamer specifically recognizing human hemoglobin, an aptamer specifically recognizing Porphyromonas gingivalis, an aptamer specifically recognizing matrix metalloproteinase-8, an aptamer specifically recognizing interleukin-6, an aptamer specifically recognizing interleukin-1β, an aptamer specifically recognizing 8-hydroxydeoxyguanosine, an aptamer specifically recognizing malondialdehyde, and an aptamer specifically recognizing gingival proteinase K.

6. A periodontitis detection kit, comprising:

a first storage cavity (40), wherein a first storage space (42) for containing the sample is formed in the first storage cavity (40), the first storage cavity (40) further comprises a pushing part (30), a first end of the pushing part (30) is positioned inside the first storage cavity (40), and a second end of the pushing part (30) extends to the outside of the first storage cavity (40); the first storage space (42) is provided with a second opening (41) for balancing air pressure;

-an extraction tube (10) comprising an extraction end (12), and an outlet end arranged at a side remote from the extraction end (12), wherein the outlet end is in communication with the first storage space (42), the extraction end (12) being sized and shaped to extend between a tooth and an adjacent gum, and the side of the extraction tube adjacent to the extraction end (12) being arranged as a capillary channel (11); the extraction pipe (10) comprises a fixed pipe fixedly connected with the first storage space (42), and a movable pipe with one end detachably connected with the fixed pipe, wherein the other end of the movable pipe is the extraction end (12);

the storage device comprises a first storage cavity (50), wherein a first storage space (52) for containing liquid is formed in the first storage cavity, the first storage space is communicated with a first storage space through a first channel, at least one air pressure balancing port is arranged in the first storage space (52), one side, close to the first storage space, of the first channel is provided with a capillary channel, the inner diameter of the first end of the first channel is smaller than that of the first storage space, the inner diameter of the joint of the first storage space and the first channel is larger than that of the second end of the first channel, and a protective sleeve is further arranged outside the first channel;

When the pushing part (30) moves along the direction away from the first storage cavity (40), the action of the pushing part (30) on the air in the extraction device can drive the sample in the capillary channel (11) and the liquid in the second storage cavity to flow towards the direction close to the first storage cavity (40).

7. The kit according to claim 6, wherein,

the extraction tube is provided with at least one section of scale mark for reading the extraction amount of the sample;

and/or the extraction end (12) is rounded.

8. The kit of claim 6, wherein the first end of the pushing portion has an outer diameter dimension that is greater than an outer diameter dimension of the second end of the pushing portion;

and/or the pushing part (30) is made of rubber material.

9. The kit of claim 6, further comprising:

a test card, and the test card comprises:

a body (60);

10. The periodontitis detection kit of claim 9, wherein the first detection zone is pre-stored with a corresponding reagent comprising: one or more of an aptamer specifically recognizing human hemoglobin, an aptamer specifically recognizing Porphyromonas gingivalis, an aptamer specifically recognizing matrix metalloproteinase-8, an aptamer specifically recognizing interleukin-6, an aptamer specifically recognizing interleukin-1β, an aptamer specifically recognizing 8-hydroxydeoxyguanosine, an aptamer specifically recognizing malondialdehyde, and an aptamer specifically recognizing gingival proteinase K.