阅读说明：本技术 一种Fish唾液自动分样检测盘及其检测方法 (Fish saliva automatic sample separation detection disc and detection method thereof ) 是由 任彪 周馨怡 田而慷 陈婧怡 任瑞阳 于晓彤 唐浩迪 余美琪 张思雨 周学东 于 2020-11-17 设计创作，主要内容包括：本发明涉及一种Fish唾液自动分样检测盘及其检测方法,解决了现有技术中唾液检测方式工序复杂、精确度不足、难以快速同时检测的问题。本发明设置基体、中央收集区、导流毛细管和玻片载台构成唾液放置、分样和放置载玻片的通道,为一体结构,其中载玻片可拆卸。将采集唾液,放入中央搜集区后,液体与毛细管壁通过毛细现象使唾液自动流入导流毛细管至玻片载台上的载玻片。拆卸载玻片后,根据Fish技术原理和检测微生物种类,判断微生物含量,指征疾病进程。多个玻片载台可以同时互不干扰检测多种微生物。实现了快速均匀的唾液自动分装,并与Fish荧光检测技术融合,特异性荧光探针与微生物核酸结合,定量检测微生物。简化唾液检测,提高唾液检测的精确度。(The invention relates to a Fish saliva automatic sample separation detection disc and a detection method thereof, and solves the problems that in the prior art, a saliva detection mode is complex in process, insufficient in accuracy and difficult to detect quickly and simultaneously. The device is provided with a substrate, a central collecting area, a diversion capillary tube and a slide glass carrier to form a channel for holding saliva, separating samples and holding a slide glass, and the channel is of an integrated structure, wherein the slide glass is detachable. After the collected saliva is placed in the central collection area, the liquid and the capillary wall automatically flow into the diversion capillary to the glass slide on the glass slide carrier through the capillary phenomenon. After the glass slide is disassembled, the content of the microorganisms is judged according to the Fish technical principle and the species of the detected microorganisms, and the disease process is indicated. Multiple slide stages can detect multiple microorganisms simultaneously without interfering with each other. The saliva is automatically subpackaged quickly and uniformly, and is fused with a Fish fluorescence detection technology, and a specific fluorescent probe is combined with the microbial nucleic acid to quantitatively detect the microbes. Simplify saliva detection, improve saliva detection's accuracy.)

1. A Fish saliva automatic sample separation detection disc, comprising: a flow guide capillary tube (3) and a glass slide carrying platform (4) are arranged in the central collecting area (2) of the substrate (1); the central collection area (2), the flow guide capillary (3) and the slide carrying platform (4) are embedded above the substrate (1).

2. The Fish saliva automatic sample separation detection disc of claim 1, wherein the central collection area (2), the flow guide capillary (3) and the slide glass carrier (4) are sequentially connected and arranged to form a saliva placement, automatic sample separation and slide glass placement channel, and the structure is an integral structure.

3. A Fish saliva automatic sample separation detection disc according to claim 2, characterized in that the flow guide capillary (3) in the central collection area (2) of the substrate (1) and the slide carrier (4) of the substrate (1) are both non-detachable.

4. A Fish saliva automatic sample separation test tray according to claim 3, wherein the slide carrier (4) is adapted for holding a slide, and the slide is freely removable.

5. A use method of a Fish saliva automatic sample separation detection disc, which is characterized in that after saliva is automatically separated and guided into a glass slide by the Fish saliva automatic sample separation detection disc of claims 1-4, the glass slide is subjected to a series of treatments and then inserted back into a glass slide carrier (4) for disposable fluorescence detection according to the principle of Fish technical steps, so as to judge the content of microorganisms and indicate the disease process, and the method comprises the following steps:

step S1: collecting a proper amount of saliva and filling the saliva into a central collecting area (2);

step S2: saliva in the central collection area (2) automatically flows into a glass slide arranged above the glass slide carrying platform (4) along the diversion capillary tube (3) through capillary phenomenon;

step S3: disassembling the glass slide, performing characteristic treatment on different glass slides according to the Fish technical step principle and the detected microorganism species, and putting specific probes into the different glass slides;

step S4: the processed slide is inserted back into the slide stage (4) and placed under a fluorescence microscope for scanning and the results are analyzed.

Technical Field

The invention relates to the field of medical technology diagnosis, in particular to a Fish saliva self-sampling detection disc and a detection method thereof.

Background

Saliva is an important body fluid secreted by the salivary glands of the oral cavity, is rich in biomolecules, and can be changed along with the physical health condition, wherein related biomarkers can be used for monitoring oral cavity and systemic diseases. Saliva detection has the advantages of being noninvasive, convenient, rapid, easy to accept by patients and the like, so that more and more attention is paid to the saliva detection. At present, the relation between various diseases such as new coronary pneumonia, oral diseases, cardiovascular diseases, cancers and the like and corresponding pathogenic bacteria and secretions thereof is clearly researched, and researches prove that the types and the proportions of flora have good correlation with the diseases. Therefore, the sensitivity and the accuracy of disease detection can be greatly improved by combining the specific microorganisms with some clinical indexes, and the detection of the abundance or the proportion change can also be used for judging the risk of diseases, helping patients to take treatment measures as soon as possible and avoiding unnecessary pain and burden caused by delay.

In addition, in recent years, the Fish technology has gradually emerged, and has the advantages of 1, economy and safety of a fluorescent reagent and a probe as a non-radioactive detection system; 2. the probe is stable and can be used within two years after being labeled once; 3. the experimental period is short, the result can be obtained quickly, the specificity is good, the positioning is accurate, and the sensitivity is high; 4. multicolor FISH can detect multiple sequences simultaneously, etc. by displaying different colors in the same nucleus.

In the prior art, the Fish saliva automatic sample separation detection disc is designed, and the saliva is automatically separated and placed on eight glass slides, so that each glass slide can be applied to detection of different microorganisms, the required saliva extraction times are greatly reduced, and the detection efficiency is improved. After the detachable glass slide is processed and the pre-configured fluorescent probe is placed in the detachable glass slide, the detachable glass slide is placed back to the device for fluorescent detection at the same time, so that various disease detection indexes can be obtained at one time, and diseases can be diagnosed efficiently and comprehensively.

Disclosure of Invention

In view of the above analysis, the present invention aims to provide a Fish saliva automatic sample separation detection tray, which is used for solving the problems of complicated procedures, insufficient accuracy, difficulty in rapid and simultaneous detection in the saliva detection mode in the prior art.

The purpose of the invention is mainly realized by the following technical scheme:

in the technical scheme of the invention, the Fish saliva automatic sample separation detection disc device comprises: the device comprises a substrate, a central collecting area, a diversion capillary tube and a slide carrying platform.

In the technical scheme of the invention, the substrate, the central searching and collecting area, the flow guide capillary tube and the slide carrying platform are of an integrated structure and cannot be disassembled.

Furthermore, the central searching and collecting area, the flow guide capillary tube and the slide glass carrying platform are sequentially arranged and embedded above the machine body to form a channel for placing saliva, separating samples and placing slide glass, and the channel is of an integrated structure.

Further, the slide stage may be for a slide to be placed.

Further, the slide is freely detachable.

A use method of a Fish saliva automatic sample separation detection disc is characterized in that after saliva is automatically separated and guided into a glass slide by the Fish saliva automatic sample separation detection disc, according to the Fish technical step principle, the glass slide is subjected to a series of treatments and then inserted back to a glass slide carrier for one-time fluorescence detection, so that the content of microorganisms is judged, and a disease process is indicated: comprises the following steps;

s1, collecting a proper amount of saliva and filling the saliva into a central collection area;

s2, automatically flowing saliva in the central collection area into a glass slide placed above the glass slide carrier along a diversion capillary tube through a capillary phenomenon;

s3, disassembling the glass slide, performing characteristic treatment on different glass slides according to the Fish technical step principle and the detected microorganism species, and putting specific probes;

and S4, inserting the processed slide back to the slide carrier, placing the slide carrier under a fluorescence microscope for scanning, and analyzing the result.

On one hand, the invention can use the Fish saliva automatic sample separation detection disc matched with probes and reagents to detect a plurality of common oral diseases, and can also use the Fish saliva automatic sample separation detection disc alone to select other microorganism specific probes and reagents to detect.

The matched probe and reagent of the invention and the application method are as follows:

slide 1 slide trough: for detecting enterococcus faecalis, the 5' end of gene probe ENF191 (sequence: GAAAGCGCCTTCACTTATGC) is marked with fluorescein isothiocyanate (green signal).

Slide groove No. 2: detection of Streptococcus mutans, Gene Probe MUT590, 5'-ACTCCAGAC TTTCCTGAC-3'.

Slide groove No. 3: the gene probe 5'-CTGGAGAGACTAAGCCCTCC-3' modified by LNA, and the 5' end Cy5 marker (Abs/Em 550-592/633 nm, red) are used for detecting helicobacter pylori.

Slide 4 slide trough: the detection of the mycobacterium tuberculosis is carried out by gene probes FUS664 and 5'-CTTGTAGTTCCGCTTACCTA-3' and 5' end Cy5 labeling (Abs/Em 550-592/633 nm, red).

Slide No. 5 slide trough: detecting Porphyromonas gingivalis, wherein a gene probe 5 '-3': CACTGAACT CAAGCCCGGCAGTTTCAA and labeled with ALEXA Fluor 488.

No. 6 slide glass slide groove, detecting Actinobacillus actinomycetemcomitans, gene probe, sequence 5'-CACCAGGGCTAAACCCAAT-3', and labeling with FITC developing blue-green.

Slide slots No. 7 and No. 8 are spare slide slots or are used for other microbiological tests.

The matched reagent comprises: fluorescent medium oil, ethanol, paraformaldehyde, sodium dodecyl sulfate, Tris buffer solution, formalin, formamide/2 XSSC (sodium dodecyl sulfate)/denaturing solution, Giemsa staining agent, PI/anti-fiade staining agent, DAPI/anti-fiade staining agent and a universal probe.

The technical scheme of the invention can at least realize one of the following effects:

1. the Fish saliva automatic sample separation detection disc simplifies operation on the basis of the original saliva detection process, realizes simultaneous detection of multiple microorganisms in the same sample, and greatly improves saliva detection efficiency.

2. The symmetrical structure of the device enables the samples to be uniformly subpackaged, and the design of mutual separation of the device areas improves the operability of the experiment, provides sufficient variable experiment conditions, fully ensures the specificity of Fish technology detection conditions, and improves the accuracy of saliva detection.

3. The device has the advantages of flexible and simple design structure, easy disassembly and cleaning and high cleaning efficiency.

4. The device disclosed by the invention is combined with a fluorescein in-situ hybridization technology (Fish), and a mode of combining a specific fluorescent probe and microbial nucleic acid is adopted to realize rapid quantitative detection of microbes.

Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The objectives and other advantages of the invention will be realized and attained by the structure particularly pointed out in the written description and drawings.

Drawings

The drawings are only for purposes of illustrating particular embodiments and are not to be construed as limiting the invention, wherein like reference numerals are used to designate like parts throughout.

FIG. 1 is a schematic top view of the present invention;

FIG. 2 is an isometric view of the present invention;

FIG. 3 is a right side view of the present invention;

FIG. 4 is a schematic view of a slide assembly of the present invention.

Reference numerals:

1-a substrate; 2-a central collection region; 3-diversion capillary; 4-slide stage.

Detailed Description

The preferred embodiments of the present invention will now be described in detail with reference to the accompanying drawings, which form a part hereof, and which together with the embodiments of the invention serve to explain the principles of the invention and not to limit its scope.

In the description of the embodiments of the present invention, it should be noted that, unless otherwise explicitly stated or limited, the term "connected" should be interpreted broadly, and may be, for example, a fixed connection, a detachable connection, or an integral connection, which may be a mechanical connection, an electrical connection, which may be a direct connection, or an indirect connection via an intermediate medium. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.

The terms "top," "bottom," "above … …," "below," and "on … …" as used throughout the description are relative positions with respect to components of the device, such as the relative positions of the top and bottom substrates inside the device. It will be appreciated that the devices are multifunctional, regardless of their orientation in space.

As shown in fig. 1 to 4, an embodiment of the present invention provides a Fish saliva automatic sample separation detection tray, including: a flow guide capillary tube (3) and a glass slide carrying platform (4) are arranged in the central collecting area (2) of the substrate (1); the central searching and collecting area (2), the flow guide capillary tube (3) and the slide carrying platform (4) are embedded above the substrate (1); the central searching and collecting area (2), the flow guide capillary tube (3) and the slide glass carrying platform (4) are sequentially connected and arranged to form a channel for placing saliva, automatically separating samples and placing slide glass, and the channel is of an integrated structure; the flow guide capillary tube (3) in the central collection area (2) of the substrate (1) and the slide carrying platform (4) are not detachable from the substrate (1); the slide glass carrier (4) can be used for placing the slide glass, and the slide glass can be freely detached. Wherein, the central collecting area (2) is a depression in the substrate (1) and is used for containing a saliva sample to be measured; the flow guide capillary tube (3) is connected with the central collecting area (2), and a part of saliva sample to be measured is guided to the slide glass carrying platform (4) by utilizing the capillary phenomenon; the slide glass carrier (4) is connected with the diversion capillary (3), a part of saliva samples are collected and then detached, different glass slides are processed differently according to the Fish technical step principle and the microbial species detection, then the slide glass carrier (4) is inserted back and placed under a fluorescence microscope for scanning, and the result is analyzed.

The invention relates to a Fish technology and an automatic sample separation technology, which are difficult to perform multiple examinations simultaneously due to the difficulty in sampling saliva and the small sampling amount in the traditional saliva detection, and have high time cost. According to the device, one saliva sample to be detected is automatically divided into eight parts on average, so that the novel saliva detection box collects the sample through the central collection area (2), the flow guide capillary tube (3) uniformly disperses the sample, and the sample is continuously drained to the slide glass carrying table (4) for subsequent operation.

After a sample to be detected is dripped in the central collecting area, liquid is in contact with the wall of the capillary tube to generate a capillary phenomenon, so that a part of liquid moves to the test paper detecting area and is absorbed by dry test paper, the sample is continuously and uniformly dispersed to each slide carrier, and the slide is disassembled for characteristic treatment and then inserted back to the slide carrier for scanning observation and result analysis.

The use method of the embodiment of the invention comprises the following steps:

step S1: collecting a proper amount of saliva and filling the saliva into a central collecting area (2);

step S2: saliva in the central collection area (2) automatically flows into a glass slide arranged above the glass slide carrying platform (4) along the diversion capillary tube (3) through capillary phenomenon;

step S3: disassembling the glass slide, performing characteristic treatment on different glass slides according to the Fish technical step principle and the detected microorganism species, and putting specific probes into the different glass slides;

step S4: the processed slide is inserted back into the slide stage (4) and placed under a fluorescence microscope for scanning and the results are analyzed.

Compared with the existing detection mode, the Fish saliva automatic sample separation detection disc provided by the embodiment of the invention has at least one of the following beneficial effects:

1. the traditional saliva test needs a large amount of saliva samples, and the testee may feel discomfort such as regurgitation during sampling. The invention can uniformly disperse the saliva sample into eight parts, reduce the times of saliva extraction and reduce the pain of the examined person.

2. One sample can be used for simultaneously detecting eight diseases, so that the detection efficiency is improved, and the time cost is saved.

3. By applying the Fish technology, the detection accuracy is improved, and the early diagnosis of the disease condition is facilitated, so that the early discovery, early diagnosis and early treatment are realized.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention.