CN111398511A

CN111398511A - Detection device and detection method for detecting helicobacter pylori

Info

Publication number: CN111398511A
Application number: CN202010260935.9A
Authority: CN
Inventors: 孙少民
Original assignee: Hangzhou Bufeng Technology Co ltd
Current assignee: Hangzhou Bufeng Technology Co ltd
Priority date: 2020-04-03
Filing date: 2020-04-03
Publication date: 2020-07-10

Abstract

The invention provides a detection device for detecting helicobacter pylori, which comprises an upper layer supporting part, a lower layer supporting part and at least two middle layer reaction parts, wherein the middle layer reaction parts are combined with the upper layer supporting part or/and the lower layer supporting part; the substances embedded on the middle layer reaction member for performing the control test comprise buffer solution and acid-base indicator, and the substances embedded on the middle layer reaction member for performing the index test comprise buffer solution, acid-base indicator and urea. The invention also provides a method for detecting helicobacter pylori. By adopting the detection device, a solid-liquid mixture, a viscous sample or a liquid sample can be tested at one time to obtain the detection result of one or more indexes; the detection method is simple and rapid, and the invention reasonably sets contrast, can eliminate interference and makes the detection result more accurate.

Description

Detection device and detection method for detecting helicobacter pylori

Technical Field

The invention particularly relates to a detection device and a detection method for detecting helicobacter pylori.

Background

Currently, there are various detection devices and detection methods for gastric helicobacter pylori, which can be roughly divided into two categories, invasive and non-invasive.

Wherein invasive means removing the gastric mucosa by gastroscopy and then examining the gastric mucosa. Including culture, smear gram staining microscope observation, tissue section staining, molecular biology detection, and rapid urease detection (rapid urea Test). Clinically RUT is the first means of invasive detection.

The non-invasive means a detection means without gastroscopy sampling, including Urea Breath Test (UBT), commonly known as 13C and 14C detection, 15N ammonia discharge Test, fecal helicobacter pylori antigen detection, helicobacter pylori antibody serology detection, urine helicobacter pylori antibody detection, urease detection of specific parts of the oral cavity and the like. Among them, 13C and 14C are widely used, are suitable for testing the current symptom infection and become a common method for clinical diagnosis in many countries.

Helicobacter pylori is consistently recognized as a high causative agent of gastric cancer. According to study statistics, approximately 50% of the world's population is currently infected with H.pylori. The above methods have some problems and are difficult to satisfy the requirement of the large base number.

Invasive methods can only be performed in hospitals with certain conditions, non-invasive tests such as 13C and 14C, 15N ammonia urine excretion tests need instruments, and serological tests need blood sample collection. These present certain difficulties.

The specific antigen of helicobacter pylori is detected in excrement, the detection of the specific antibody in urine needs expensive immunological technology as support, and compared with a urease biochemical method, the detection time is long and the operation is complex.

Urease detection of specific parts of the oral cavity is a better option.

First, the oral cavity is the repository for H.pylori. The mouth is located at the front end of the digestive tract and is in direct physical communication with the stomach. The presence of H.pylori has been reported in saliva, dental plaque, tongue dorsum, oral mucosa, root canal and tonsil. Failure of antibiotics to eradicate H.pylori may have some correlation with H.pylori in the oral cavity, and oral transmission has been discovered. These all indicate that the oral cavity can be used as the site of the digestive tract for testing H.pylori.

Second, oral sampling is very easy, not relying on excessive technical means for support.

However, sampling at specific positions is different, for example, dental plaque can be safely carried out under the operation of a professional physician by using a hard instrument, tonsils are deep in the oral cavity, and sampling is easy to cause discomfort; the root canal is easy to be carried out only when the decayed tooth is treated; saliva has a limited and unstable helicobacter pylori bacterial content.

The tongue coating on the back of the tongue is the better choice. The tongue coating refers to the dirty coating on the surface of the back of the tongue, and consists of tongue mucosa cells, exfoliated cells, exuded leukocytes, saliva, bacteria, fungi and physical debris. The components of the tongue coating have certain stability. The tongue coating can be scraped in a sufficient amount by a proper way to be used as a detection sample, so the detection is very suitable.

The tongue coating is relatively complicated in composition and is affected by diet. The simple urease test may be disturbed, resulting in inaccurate results. The currently marketed urease detection device does not consider the complexity of a sample, does not set a control, has inaccurate detection results, is easy to generate false positive or false negative results, and is inconvenient to use.

The commonly used method is the colloidal gold method, and therefore also many detection devices are produced. The colloidal gold method generally detects an antigen or an antibody, the antigen sample is feces, and the antibody sample is blood, serum, fingertip blood, or the like. The method has the advantages of simplicity, convenience, high accuracy and no limitation of conditions. However, the colloidal gold method requires 8 minutes for measurement, requires a specific antigen, a secondary antibody, a labeling substance, and the like, and has a complicated detection apparatus structure.

Disclosure of Invention

The invention aims to solve the technical problem in the background art and provides a detection device and a detection method for detecting helicobacter pylori.

Therefore, the technical scheme adopted by the invention is as follows:

a detection device for detecting helicobacter pylori, comprising an upper layer support member, a middle layer reaction member, a lower layer support member, wherein the middle layer reaction member is combined with the upper layer support member or/and the lower layer support member, the detection device comprises at least two middle layer reaction members, wherein at least one middle layer reaction member can be used for carrying out contrast test, and other middle layer reaction members can be used for carrying out detection of at least one index; the substances embedded on the middle layer reaction member for performing the control test comprise a buffer solution and an acid-base indicator, and the substances embedded on the middle layer reaction member for performing the index test comprise the buffer solution, the acid-base indicator and urea.

Further, the substances embedded on the middle layer reaction member for performing the control test comprise a buffer solution, an acid-base indicator and a preservative, and the substances embedded on the middle layer reaction member for performing the index test comprise a buffer solution, an acid-base indicator, a preservative and urea.

Further, the buffer solution includes, but is not limited to, at least one of glycine buffer solution, citric acid buffer solution, phosphoric acid buffer solution, morpholine ethanesulfonic acid buffer solution at pH 2.0-7.0.

Further, the acid-base indicator includes, but is not limited to, at least one of cresol red, phenol red, bromocresol purple.

Further, the preservative includes sodium benzoate, sodium azide, Proclin 300.

Further, the substances dissolved in the aqueous solution embedded on the middle layer reaction for the control test: 0.01-0.1M citric acid buffer pH3.0-pH6.0, 0.3-30mM phenol red and 0.05-5mM sodium benzoate; substances dissolved in aqueous solution embedded on the middle layer reaction piece for index test: 0.01-0.1M citric acid buffer pH3.0-pH6.0 contains 0.3-30mM phenol red, 0.05-5mM sodium benzoate and 0.5-500mM urea.

Furthermore, one end of the upper layer supporting piece is connected with one end of the lower layer supporting piece; the lower support or the upper support is provided with a sample adding region.

A method for detecting helicobacter pylori, which uses the detection device described above, comprising the steps of:

(1) opening the upper layer supporting piece to enable the upper layer supporting piece and the lower layer supporting piece to be tiled on the same plane;

(2) respectively adding 1mg-10mg of sample to the control group sample adding area and the experimental group sample adding area of the lower layer supporting piece;

(3) after the sample is loaded, the upper layer supporting piece is turned back, and the upper layer supporting piece and the lower layer supporting piece are combined together;

(4) flattening to enable the sample loaded on the lower layer supporting member to penetrate into the middle layer reaction member;

(5) and starting timing, and observing the color change of the middle layer reaction member.

Further, in the step (3), the sample is a tongue coating sample.

Further, in the step (5), the color change of the middle layer reaction member is observed within 0.5 to 5 minutes.

The invention has the beneficial effects that:

(1) the invention is a dry biochemical reaction detection device based on the specificity of biological enzyme and the principle of high-efficiency catalysis; by adopting the detection device, a solid-liquid mixture, a viscous sample or a liquid sample can be tested at one time under the condition of not depending on an instrument, and the qualitative detection result of one or more indexes can be obtained; the detection device can be applied to the fields of medicine, veterinary medicine, agriculture, animal husbandry, food safety, environmental monitoring, biological safety and the like.

(2) The detection device is provided with the middle-layer reaction member (control group) for carrying out control test, and the control group can be added for detecting the same sample during detection, so that interference is eliminated, and the detection result is more accurate.

(3) The detection device can ensure that the analyte liquid is partially diffused by extrusion to promote the biochemical reaction to occur, and the detection result is quickly obtained. The detection device of the invention has two or more middle-layer reaction parts fixed in different areas of the upper-layer support part or/and the lower-layer support part, and a certain physical distance exists between the two reaction parts to ensure that the reaction parts are mutually independent and do not interfere with each other. Thus, one index detection can be performed on one sample or multiple index detections can be performed on one sample. The same sample is loaded in the reaction areas with different indexes, so that the detection results of various indexes can be obtained.

(4) The end part of the upper layer supporting piece and the end part of the lower layer supporting piece are connected together by adopting a hinge structure; when the upper layer supporting piece needs to be opened, the upper layer supporting piece can be turned over when the sample is added, the upper layer supporting piece is naturally unfolded on the plane, the middle layer reaction piece and the sample adding area are exposed, so that two hands of a user can be conveniently vacated, a sample can be smoothly added in the sample adding area, after the sample addition is completed, the upper layer supporting component and the lower layer supporting component are buckled at the original position, and the subsequent result can be conveniently interpreted.

(5) According to the invention, the sample adding region is arranged on the lower supporting piece or the upper supporting piece, and the arrangement of the sample adding region is beneficial to accurately and quickly adding a sample to the detection device, so that the time is saved, and the efficiency is improved. The sample adding region and the middle layer reaction member are usually arranged on different supporting members, thereby being beneficial to sample adding and not causing the pollution of samples. If the sample adding region and the middle layer reaction piece are both arranged on the same supporting piece, namely the positions of the sample adding region and the middle layer reaction piece are overlapped, substances embedded on the middle layer reaction piece can be touched in the sample adding process, so that the pollution condition can exist possibly, the accuracy of a control experiment is influenced, and the sample adding of a control group and the sample adding of an experiment group are performed at certain time.

(6) The detection method of the invention needs less sample amount, is simple to operate and can quickly obtain the detection result; the invention reasonably sets contrast, can eliminate interference and leads the detection result to be more accurate.

Drawings

FIG. 1 is an exploded view of the detection device.

Fig. 2 is a schematic structural diagram of the detection device.

Fig. 3 is a schematic view of the upper support member in an open configuration.

FIG. 4 is a schematic view showing a structure in which a specimen is applied to a middle reaction member.

FIG. 5 is a graphical representation of the results of a valid positive test in the absence of interference.

FIG. 6 is a graph showing the results of effective negative tests in the absence of interference.

FIG. 7 is a diagram illustrating one of the invalid detection results in the presence of interference.

FIG. 8 is a schematic diagram of another invalid detection result in the presence of interference.

Fig. 9 is a schematic diagram of the structure of the sampler in embodiment 2.

FIG. 10 is a schematic diagram showing the structure of the sample injector in example 3.

FIG. 11 is a schematic diagram showing the structure of the sampler in combination with the sampler in example 3.

Fig. 12 is a schematic side view showing the structure of the sampler combined with the sampler in example 2 (showing the structure of the first handle).

FIG. 13 is a schematic view of the structure of the sampler in example 4.

FIG. 14 is a schematic diagram showing the structure of the sample injector in example 5.

FIG. 15 is a schematic diagram showing the structure of the sampler in combination with the sampler in example 5.

FIG. 16 is a schematic view showing a configuration in which an upper support is expanded in one embodiment of the detecting unit of example 1.

FIG. 17 is a schematic diagram showing a positive test result in one configuration of the test device in example 1.

FIG. 18 is a schematic view showing a structure in which an upper support is expanded in another configuration of the detecting unit according to embodiment 1.

FIG. 19 is a schematic view showing the result of negative detection in another configuration of the detecting unit in example 1.

FIG. 20 is a schematic diagram showing a result of a positive test in another configuration of the test device in example 1.

FIG. 21 is a schematic diagram showing another arrangement of the sample injector in example 3.

Detailed Description

The following detailed description of the embodiments of the present invention is provided in conjunction with the accompanying drawings, and it should be noted that the embodiments are merely illustrative of the present invention and should not be construed as limiting the present invention.

Example 1, reference is made to FIGS. 1-8, 16-20.

1-2, a biochemical reaction-based detection device comprises an upper layer support member 1, a middle layer reaction member 2, a lower layer support member 3, wherein the middle layer reaction member 2 is combined with the upper layer support member 1 or/and the lower layer support member 3, and in some preferred modes, as shown in FIG. 3, the middle layer reaction member 2 is combined and fixed with the upper layer support member 1; in some preferred embodiments, the detection device comprises at least two middle-layer reaction members 2, wherein at least one of the middle-layer reaction members 2 can be used for performing a control test, and the other middle-layer reaction members 2 can be used for performing a detection of at least one index. More than one middle-layer reaction part 2 can be arranged for contrast test, so that interference can be eliminated better, invalid results can be identified, and the detection result is more accurate; moreover, the device of the invention can also adopt a plurality of middle-layer reaction pieces 2, thereby being capable of carrying out detection on a plurality of indexes of the sample.

In some preferred modes, the middle reaction member 2 can perform some biochemical reactions, and in some preferred modes, the middle reaction member 2 is embedded with substances and related auxiliary agents participating in the biochemical reactions in advance, and the embedded substances can react with substances to be detected in a sample, so that the existence of a certain substance can be detected. In some preferred modes, all the substances and related auxiliary agents participating in biochemical reactions are embedded in the middle layer reaction member 2 (i.e. experimental group) for performing certain index detection in advance; the middle layer reaction member 2 (i.e. the control group) for performing the control test is pre-embedded with a part of the substances participating in the biochemical reaction and the related auxiliary agents, and the non-embedded substances are necessary and insufficient conditions for detecting the analyte. In some preferred modes, the chemical composition components embedded on the middle layer reaction member 2 (control group) are at least one less than the chemical composition components of the middle layer reaction member 2 (experimental group), wherein the same component chemical has the same amount.

In some preferred modes, the substance embedded on the middle layer reaction member for performing the control test comprises a buffer solution and an acid-base indicator, and the substance embedded on the middle layer reaction member for performing the index test comprises a buffer solution, an acid-base indicator and urea. In some preferred modes, the substances embedded on the middle layer reaction member for performing the control test comprise a buffer solution, an acid-base indicator and a preservative, and the substances embedded on the middle layer reaction member for performing the index test comprise a buffer solution, an acid-base indicator, a preservative and urea.

In some preferred embodiments, as shown in FIGS. 16-20, the lower support (or the upper support) is provided with indicia identifying the middle reaction part 2 (control group) and the middle reaction part 2 (experimental group), such as C for the control group and T for the experimental group, where there are multiple experimental groups, T1, T2, T3 …. Tn, or other symbols. When there are a plurality of control groups, they may be represented by C1, C2, C3 …, Cn, or by other different symbols.

In some preferred modes, the upper layer support 1 or the lower layer support 3 mainly supports and fixes, for example, the middle layer reaction part 2 can be fixed on the upper layer support 1 (or the lower layer support 3), and when sample is added, the sample can be added on the middle layer reaction part 2, as shown in fig. 4; the sample can react with the substance embedded in the middle reaction member 2 in advance; or the sample is added to the position corresponding to the lower layer support 3 (or the upper layer support 1) and the middle layer reaction member 2, for example, the sample adding region 10 can be pre-printed at the corresponding position on the lower layer support, as shown in fig. 18, the circle region in fig. 18 is the sample adding region 10, the sample can be added to the sample adding region 10, the arrangement of the sample adding region 10 is beneficial to quickly and accurately adding the sample to the detection device, the time is saved, and the efficiency is improved, so that when the upper layer support 1 and the lower layer support 3 are combined, the sample can be in contact with the middle layer reaction member 2, the sample can permeate into the middle layer reaction member 2, and the sample and the substance embedded in the middle layer reaction member 2 perform biochemical reaction.

In some preferred modes, the sample application region 10 is located at the middle position in the width direction of the lower layer support, which is beneficial for sample application and can avoid the sample from overflowing the edge of the lower layer support due to improper sample application position. In some preferred modes, the sample application region and the middle layer reaction member are usually disposed on different supports, for example, if the middle layer reaction member is combined with the upper layer support 1, the sample application region is disposed on the lower layer support at a position corresponding to the middle layer reaction member; if the middle layer reaction member is combined with the lower layer support member 1, the sample adding region is arranged on the upper layer support member at a position corresponding to the middle layer reaction member; the sample adding region and the middle layer reaction piece are arranged on different supporting pieces, so that sample adding is facilitated, sample pollution can not be caused, and sample pollution is easily caused if direct sample adding is carried out on different middle layer reaction pieces, because substances embedded on different middle layer reaction pieces are possibly different, for example, the middle layer reaction piece (a control group) and the middle layer reaction piece (an experimental group) or the middle layer reaction pieces embedded with different substances for detecting different indexes.

In some preferred embodiments, two or more middle reaction members 2 are fixed to different regions of the upper support member 1 or/and the lower support member 3 at a physical distance from each other to ensure independence from each other and non-interference. Thus, one index detection can be performed on one sample or multiple index detections can be performed on one sample. In some preferred modes, as shown in fig. 16, an adhesive is used to fix the upper support member 1 (or the lower support member 3) and the middle reaction member 2 together.

In this embodiment, as shown in fig. 1-2, the detecting device includes two middle layer reaction members 2, and the two middle layer reaction members 2 are fixed in different areas of the upper layer supporting member 1, and have a certain physical distance therebetween without interfering with each other. One of the middle layer reaction members 2 is used for contrast test, and the other one is used for detecting a certain index of a certain sample, wherein the sample can be a liquid sample, such as blood, urine, sweat, spinal fluid, saliva and the like, and the sample can also be a high-viscosity liquid sample, such as sputum, nasal mucus and secretion and the like; the sample can also be a solid-liquid mixed sample, such as a tissue sample, a bacterial colony, a fecal sample, a culture, tartar, plaque, tongue coating, and the like; the sample may also be a solid sample, such as soil, with no or low liquid content. The indicator may be the detection of a certain bacterium (e.g. helicobacter pylori) or a certain virus or the detection of a certain antibody or the detection of a certain chemical substance, such as a drug, etc.

In some preferred forms, one end of the upper support member 1 is connected to one end of the lower support member 3. In some preferred modes, one end of the upper layer supporting member 1 and one end of the lower layer supporting member 3 are connected together by a hinge structure. In some preferred forms, one end of the upper support member 1 is joined to one end of the lower support member 3 by a single-sided adhesive. One end of the single-sided adhesive is connected with the outer side of the upper layer supporting member 1, and the other end is connected with the outer side of the lower layer supporting member 3. In some preferred forms, the single-sided adhesive has a thickness of between 0.1 and 0.2 mm. In this embodiment, as shown in fig. 3-4 and 16, a portion of the single-sided adhesive thin layer is connected to the outer side (i.e. upper side) of the upper layer supporting member 1, and another portion of the single-sided adhesive thin layer is connected to the outer side (i.e. lower side) of the lower layer supporting member, so that the single-sided adhesive thin layer itself forms a "hinge region", when the upper layer supporting member 1 needs to be opened for sample application, the upper layer supporting member 1 can be turned over to naturally spread the upper layer supporting member 1 on a plane, as shown in fig. 3 and 16, the middle layer reaction member 2 fixed on the inner side of the upper layer supporting member is exposed, so that a user can conveniently remove both hands to smoothly apply a sample to each middle layer reaction member 2 or apply a sample to a position corresponding to the middle layer reaction member on the lower layer supporting member, after sample application is completed, the upper layer supporting member and the lower layer supporting member are fastened together, and the sample is in contact with the middle layer reaction member, the reaction is carried out, and the subsequent detection result can be conveniently interpreted. In some preferred forms, the lower support member and the upper support member are secured together by a flexible adhesive tape, as shown in figure 18. The flexible adhesive tape connects the ends of the upper and lower support members and can function as a hinge.

In some preferred modes, the length of the non-connecting end of the upper layer support 1 and the lower layer support 3 is different, so that the upper layer support 1 can be easily separated from the non-connecting end, and the middle layer reaction member and the sample adding area 10 can be exposed, so that the sample can be conveniently added. In some preferred modes, corresponding marks are arranged at the connecting ends of the upper-layer supporting member 1 and the lower-layer supporting member 3, or different colors are arranged to distinguish the connecting ends and the non-connecting ends, so that the connecting ends and the non-connecting ends can be conveniently identified and recognized. As shown in fig. 18, the connecting ends of the upper layer supporting member 1 and the lower layer supporting member 3 are provided with a darker color to distinguish the connecting ends from the non-connecting ends, and for example, the connecting ends may be provided with a color such as red or green.

In some preferred forms, the upper support 1 has a thickness of between 0.1mm and 1mm or the lower support 3 has a thickness of between 0.1mm and 1 mm. Thus being convenient for production and operation by users.

In some preferred forms, the upper support member 1 is made of a transparent plastic material, including but not limited to PE (polyethylene), PP (polypropylene), ABS (acrylonitrile/butadiene/polystyrene copolymer), PC (polycarbonate), PET (polyethylene terephthalate), PVC (polyvinyl chloride), PS (polystyrene), P L A (polylactic acid resin).

In some preferred forms, the lower support member 3 is a white or light colored plastic sheet, including but not limited to polyethylene, polyvinyl chloride, polypropylene, polycarbonate, and the like. In some preferred forms, at least the inner side of the lower support member is sanded. This is beneficial to the sample loading area of the lower layer supporting member, and the sample is prevented from overflowing and flowing around.

In some preferred modes, a layer of easily-torn self-adhesive sticker can be not attached or pre-attached to the lower-layer supporting member 3, so that the operation of an operator is facilitated or a test result is kept.

In some preferred modes, the middle reaction member 2 is made of porous material. The sample can be kept by the porous material, and the sample is easily contacted with the middle layer reaction piece 2, so that the sample and substances embedded on the middle layer reaction piece can be favorably reacted, and the liquid sample can be prevented from overflowing out of the testing device. In some preferred forms, the intermediate reaction member is a porous hydrophilic material comprising cellulose-based filter paper containing-CONHCH₂OH、-CONH₂、-COOH、-COOROH、-NH2、-OH、-SO₃H, and the like. The porous hydrophilic material can absorb water in the sample, and the sample containerIs easy to contact with the middle layer reaction member 2, and is beneficial to the reaction between the sample and the substance embedded on the middle layer reaction member. In some preferred forms, the porous material includes, but is not limited to, filter paper, polyester, fiberglass. In some preferred forms, the porous material has a thickness of between 0.1mm and 1 mm.

The using method of the detection device comprises the following steps: (As shown in FIGS. 3 to 4, the middle reaction member 2 is bonded to the upper support member 1, and the description will be made by way of example)

(1) Opening the upper layer supporting member 1 to make the upper layer supporting member 1 and the lower layer supporting member tiled on the same plane, as shown in fig. 3;

(2) taking a certain amount of sample, firstly adding the sample on the middle layer reaction member 2 of the control group, and then adding the sample on the middle layer reaction member 2 of the experimental group, as shown in figure 4; in some preferred modes, the sample is placed in the middle of the layer reaction member 2 in the control group or the experimental group respectively; in other preferred embodiments, the sample is applied to the lower support 3 at a position corresponding to the middle reaction member 2 (control group and experimental group);

(3) combining the upper layer supporting member 1 and the lower layer supporting member 3, extruding to tightly combine the sample and the middle layer reaction member 2, and observing color change within 0.5-5 minutes; through the extrusion, the liquid part can be diffused relatively fast in the sample, and the material that awaits measuring can fully contact with the material of embedding in advance on the middle level reaction piece 2, promotes biochemical reaction and takes place.

The detection results are usually three, the first, as shown in fig. 5, is a valid positive result in the absence of interference, the control group 2b has no color change, and the experimental group 2c has a color change; second, as shown in fig. 6, there was no effective negative result in the presence of interference, no color change in control group 2b, and no color change in experimental group 2 c; third, as shown in FIGS. 7-8, there was an invalid result in the presence of the interference, and the control group 2b had a color change.

The application of the detection device based on biochemical reaction in the detection of helicobacter pylori in the tongue fur sample is the detection device.

In some preferred embodiments, the substance embedded in the middle layer reaction member 2 (i.e., the control group) for performing the control test includes a buffer solution with a pH of 2.0-7.0 at 0.01-0.1M, and the buffer solution constituting the acid-base equilibrium includes, but is not limited to, glycine buffer solution, citric acid buffer solution, phosphoric acid buffer solution, morpholine ethanesulfonic acid buffer solution, and the like. To this solution is added 0.03-300mM acid-base indicators including, but not limited to, cresol red, phenol red, bromocresol purple, and the like. Adjuvants, such as solubilizing chemically inert substances, bacterial or mold inhibitors, etc., may also be added to the solution. The substances embedded in the middle layer reaction member 2 (i.e., experimental group) for performing the detection of a certain index are: adding 0.05mM-500mM urea on the basis of the above control group; that is, the other components in the experimental group, and the concentrations and contents thereof were the same as those in the control group except for urea.

In some preferred embodiments, the aqueous solution in which the middle layer reaction member (control) is embedded contains 0.01-0.1M citric acid buffer pH3.0-pH6.0 containing 0.3-30mM phenol red (indicator) and 0.05-5mM sodium benzoate (preservative), and the aqueous solution in which the middle layer reaction member (experimental) is embedded contains 0.01-0.1M citric acid buffer pH3.0-pH6.0 containing 0.3-30mM phenol red (indicator), 0.05-5mM sodium benzoate (preservative), and 0.5-500mM urea.1M 1 mol/L described above.

In some preferred forms, the volume of the solution is calculated in terms of the water-holding capacity of the porous material, e.g. 1cm with a thickness of 0.3mm²The porous material can carry about 20 mu L aqueous solution, the solution is embedded on the middle layer reaction member, and the specific embedding method is that the area of the porous material is calculated, corresponding amount of the solution is sprayed on the porous material, and the porous material is dried for about 12 hours at the relative humidity of less than 30 percent and the temperature of 2-40 ℃.

Principle of urease detection: helicobacter pylori is the bacterium with the highest urease content discovered at present (which is the result of the long-term evolution of the bacterium and can enable helicobacter pylori to continuously survive under acidic conditions), and the urease can catalyze urea to form ammonia under acidic conditions, so that the pH value around the urease is increased and the color of an acid-base indicator is changed along with the increase of the concentration of the ammonia. The existence of urease can be judged by the change of the color of the acid-base indicator.

The above information transfer chain can be represented as follows:

① alkali indicator color change->② increase in pH value>③NH₃Increase->④ urease Presence->⑤ helicobacter pylori is present.

The tongue coating is relatively complex and affected by diet, the simple urease detection may be interfered and result may be inaccurate, and ①② in the information transmission chain may be affected by non-urease factor.

The control settings were set to exclude those non-urease factors.

If the difference between the set control group and the experimental group is that whether the indicator exists or not, other conditions are consistent; if the pH value of the oral cavity is higher than 7.0 and the pH value of the tongue coating sample is higher than 7.0 due to the fact that the tested person drinks tea and the like, the color of the tongue coating sample is still not changed because the control group does not contain the indicator at the moment; the color of the indicator on the layer reaction member in the experimental group changes, and it is difficult to distinguish: whether the indicator color change is caused by H.pylori or the sample itself has an effect on the indicator, the setting of the control group and the test group is likely to result in false positive results.

The difference between the control group and the experimental group is whether urea exists or not, and other conditions are consistent; if the pH value of the oral cavity is higher than 7.0 and the pH value of the tongue coating sample is higher than 7.0 due to the fact that the tested person drinks tea and the like, the color of the indicator of the control group is changed when the tongue coating sample is detected, and the interference exists; if the color of the indicator on the layer reaction member in the experimental group also changes, but the red color of the experimental group exceeds that of the control group, a correct judgment can be made: the presence of H.pylori in the tongue coating samples, if not clearly distinguished, is likely to be negative, and H.pylori is absent from the tongue coating samples. This shows that the control group and the experimental group provided by the present invention can more accurately detect the presence of helicobacter pylori in the tongue coating sample.

The detection device is used for detecting whether helicobacter pylori exists in the tongue fur sample, and the specific detection method comprises the following steps: (As shown in FIGS. 18 to 19, the middle reaction member 2 is bonded to the upper support member 1, and the description will be made by way of example)

(1) The upper layer supporting piece 1 is turned over around the hinge, then the middle layer reaction piece 2 is separated from the lower layer supporting piece 3, and the upper layer supporting piece 2 and the lower layer supporting piece 3 are tiled on a plane;

(2) adding the tongue fur sample into a control group sample adding area and an experimental group sample adding area of the lower layer supporting piece 3 respectively, and adding about 1mg-10mg of the tongue fur sample into each sample adding area; the sample amount is set mainly according to the volume of water contained in the middle layer reaction piece, and the water contained in the tongue coating sample of 1-10 mg enables the whole sample to permeate into the middle layer reaction piece for reaction, so that the result can be observed and the whole sample cannot overflow out of the lower supporting piece.

(3) After the sample is loaded at the designated position on the lower layer supporting piece, the upper layer supporting piece 2 is turned back, and the upper layer supporting piece 2 and the lower layer supporting piece 3 are combined together;

(4) flattening to allow the sample loaded on the lower support member 3 to penetrate into the middle reaction member 2;

(5) at the start of the timing, the color change of the middle layer reaction member (control group) and the middle layer reaction member (experimental group) was observed.

In some preferred embodiments, the timer may be set at 0.5 to 5 minutes, within which time the observation of the color change is performed.

When the sample is observed to permeate into the middle reaction part through the transparent upper layer supporting part and the color of the middle reaction part (control group) is not changed within 0.5-5 minutes, the test of the sample is not interfered by the non-urease, and the result is accurate. Within 0.5-5 minutes, when the color of the middle reaction part (control group) is not changed, the color of the middle reaction part (experimental group) is changed, which can be regarded as a positive result, and helicobacter pylori exists. Within 0.5-5 minutes, neither the middle layer reaction member (control group) nor the middle layer reaction member (experimental group) showed color change, and the result was considered as negative. When the sample is observed to permeate into the middle reaction part through the transparent upper layer supporting part and the color of the middle reaction part (control group) changes within 0.5-5 minutes, indicating that the interference substance exists, comparing the color change difference between the middle reaction part (control group) and the middle reaction part (experimental group), if the color change is not obviously different, terminating the detection, finding out the interference substance possibly existing in the tongue coat sample, and testing after removing the interference substance; if the color change of the intermediate reaction member (experimental group) exceeds the color change of the intermediate reaction member (control group), this may be considered as a positive result.

In some preferred modes, the result can be determined by visual observation or by using an instrument.

In some preferred embodiments, semi-quantitative or quantitative results may be obtained if the sample is homogeneous and the amount loaded can be controlled within a certain range, such as ± 10%. 10% refers to the actual variation from the sample requirement for loading, if the sample requirement is 3mg, the actual sample addition is not necessarily 3mg, and the actual loading is controlled at 2.7-3.3mg, in which case the effect of the sample addition error can be eliminated, and the result is only related to a uniform sample, and the "concentration" of bacteria is considered to be the only variable for color change, so there is a semi-quantitative or quantitative basis.

In some preferred modes, if the monitoring is performed by using an instrument, the interpretation standard of the result can be the time required for reaching a certain gray level, or the result can reach a certain gray level in a fixed time.

In some preferred modes, if the test result is interpreted by using an instrument, the gray scale and the content of helicobacter pylori are in positive correlation within a certain time; the time required to reach a certain grey scale is inversely related to the bacterial content of H.pylori.

In some preferred modes, the result can be determined and explained by using the existing mobile equipment such as a mobile phone and the like and by installing corresponding application software and matching with the camera shooting function of the mobile equipment.

Example 2, see figures 9, 11, 12.

A sample acquiring device, as shown in fig. 9, comprising a sampler 1a for acquiring and/or transferring a sample from a biological tissue or organ, the sampler 1a comprising a sampling head 3a, the sampling head 3a having at least one first sample binding area 10 a. In some preferred modes, the sampling head 3a can be used for collecting samples, the sampling head 3a can be used for scraping or other modes to separate the samples from the biological tissues or organs and gather the samples in the first sample combining area 10a, and the first sample combining area 10a can combine and hold the samples, so that the samples cannot be separated from the sampling head 3a and fall elsewhere; but also allows other devices to remove the sample from the first sample binding region, the binding being reversible. In some preferred embodiments, the sampling head 3a can collect a sample from a tissue or an organ having a soft surface, where the sample can be a liquid sample or a solid-liquid mixture sample, or a sample having a certain viscosity, such as a tongue coating sample scraped from the surface of the tongue. In some preferred forms, the sampler 1a can also be used to transfer the collected sample to the detection device.

The term "bind" as used above means that the sample is brought into contact with the binding region, and the sample is held in the binding region by bringing the surface tension of the liquid sample or the solid-liquid mixed sample into contact with the surface of the binding region, or by bringing a viscous component contained in the liquid sample or the solid-liquid mixed sample into contact with the surface of the binding region. The area has a relatively large surface area and thus a large holding force for the sample, allowing the sample to stably pool in the area. Such a bonding area may be in the form of various structures, such as a bent area or a broken line area of a bent or broken line, which is illustrated with respect to a straight line form. The curved or dog-leg region has an increased relative surface area and a greater surface area for contact with the sample. In addition to the above, the surface area may be increased by roughening the portion contacting the sample, or by roughening the curved region-binding surface, or by roughening the broken line-type region-binding surface, or the like. If the sample binding regions are simply in a linear or planar configuration, such binding regions have less contact with the surface of the sample, then the binding regions bind or pool relatively less sample, particularly for liquid samples, because the liquid sample has some fluidity.

In some preferred forms, the first sample bonding area 10a includes a curved line structure or/and a broken line structure. The curved line structure and/or the broken line structure facilitate the sample to be combined on the first sample combination area 10a, such combination area is in more contact with the surface of the sample, the sample is combined on the combination area through the force of surface tension, attraction, viscosity and the like, for example, the liquid sample can be combined on the first sample combination area 10a through the surface tension. In some preferred modes, the radian of the arc is greater than 0 and less than or equal to 2 pi, and when the radian is equal to 2 pi, the arc structure is a circular ring; the arc structure can be an arc or a circular ring structure, the liquid has surface tension, and the liquid sample can be easily combined at the arc structure and is not easy to fall off. The broken line structure comprises a closed broken line and an open broken line, wherein the closed broken line is a polygonal structure, such as a triangle, a quadrangle, a pentagon and the like. In some preferred modes, the fold line structure further comprises a fold surface; the broken line structure all has the contained angle, the pitch arc structure has the fillet, and the surface contact of such combination region and sample is more, and liquid sample or solid-liquid mixture sample or the sample that has certain stickness combine in contained angle and fillet department easily. In some preferred forms, the first sample bonding area 10a includes both an arc line structure and a meander line structure. In this embodiment, the structure of the sampling head is as shown in fig. 9, the first sample combining area 10a is a triangular ring structure, and a relatively large number of liquid samples, solid-liquid mixed samples or samples with certain viscosity are easily combined at the included angle of the triangular ring structure. Fewer samples were bound at the middle of the three straight sides of the trilateral loop or at the middle of the three planes in which the three straight sides lie, such as at the 11b position. If the volume of the sampling head is smaller in the embodiment, the sample is combined in the whole trilateral annular structure, but the volume of the sampling head is too small, so that the sampling amount is limited. The sampling head is usually made to have a moderate volume, and a relatively large amount of liquid samples or solid-liquid mixed samples or samples with certain viscosity can be easily collected at the included angle. The trilateral annular structure is convenient for a user to scrape a sample from a biological tissue or an organ, the sample is easy to collect in the combination area, and the trilateral annular structure is stable and is not easy to damage when in use. In some preferred forms, each corner of the polygonal configuration is rounded as shown in fig. 9, which makes the sampler safer to use, less prone to scratching of organs or tissues, and facilitates stable incorporation or retention of the liquid sample.

In some preferred forms, as shown in fig. 9, the sampler further comprises a first handle 8a, the first handle 8a being connected to the sampling head 3 a. The first handle 8a facilitates access to the sampler. The first handle 8a and the sampling head 3a may be fixedly connected or detachably connected, for example, the first handle 8a and the sampling head 3a are connected by a snap structure.

In some preferred modes, the sampler 1a is made of a material with certain hardness, and the sampling head 3a and the first handle 8a have certain hardness, so that the sampling head can keep a certain shape, is not easy to deform, facilitates the transmission of force, and can well scrape a sample from biological tissues or organs with soft surfaces.

In some preferred modes, the sampler further comprises a connecting part 4a, as shown in fig. 9, the connecting part 4a can strengthen the connection between the sampling head 3a and the first handle 8a, so that the connection between the sampling head 3a and the first handle 8a is firmer, the connection between the sampling head 3a and the first handle 8a is prevented from being broken, the force transmission is facilitated, and the sample scraping is facilitated.

In some preferred forms, as shown in fig. 9 and 11, the connecting piece has a square end 7a with a large width, which not only facilitates the connection with the sampler, but also facilitates the taking of the sampler, so that it is not easy to fall off.

In some preferred forms, as shown in fig. 12, the first handle 8a has a gradually increasing height from the head to the tail of the first handle 8a (i.e., in the direction of the arrow in fig. 12), the first handle 8a has a lower head height to facilitate the insertion of the tongue coating, and the first handle 8a has a higher tail height to facilitate the removal of the sampler.

The above described collection device can be used to collect a tongue coating sample. Further, the collection device can be used to detect helicobacter pylori in a tongue coating sample.

In some preferred modes, the sampler is produced by one-time injection molding, the material is a high molecular polymer containing nitrogen and/or oxygen and/or chlorine and/or sulfur, and the material includes but is not limited to ABS (acrylonitrile/butadiene/polystyrene copolymer), PC (polycarbonate), PET (polyethylene terephthalate), PVC (polyvinyl chloride), PS (polystyrene), P L A (polylactic acid resin).

Example 3, see figures 10-11, 21.

In some preferred modes, as shown in fig. 10, the sample collection device further comprises an applicator 2a for collecting and transferring the sample. The sample applicator 2a can be used to apply a sample to the detection device.

In some preferred embodiments, the sample applicator 2a includes a sample applicator head 5a, and the sample applicator head 5a can be used for collecting and transferring samples, for example, the sample applicator head 5a can collect samples from the sample applicator head 3a and can transfer the samples to the detection device.

In some preferred embodiments, the sample addition head has at least one second sample binding region 11a, as shown in FIG. 10. The second sample binding region 11a can bind and hold the sample, so that the sample does not separate from the sample addition head 5a and fall elsewhere; but also under conditions allowing the sample to leave the second sample binding area 11a, where the binding is reversible. The term "bind" as used above means that the sample is brought into contact with the binding region, and the sample is held in the binding region by bringing the surface tension of the liquid sample or the solid-liquid mixed sample into contact with the surface of the binding region, or by bringing a viscous component contained in the liquid sample or the solid-liquid mixed sample into contact with the surface of the binding region. The second sample binding region has a relatively large surface area and thus a large holding force for the sample, allowing the sample to stably pool in the region. Such a bonding area may be in the form of various structures, such as a bent area or a broken line area of a bent or broken line, which is illustrated with respect to a straight line form. The curved or dog-leg region has an increased relative surface area and a greater surface area for contact with the sample. In addition to the above, the surface area may be increased by roughening the portion contacting the sample, or by roughening the curved region-binding surface, or by roughening the broken line-type region-binding surface, or the like. If the sample binding regions are simply in a linear or planar configuration, such binding regions have less contact with the surface of the sample, then the binding regions bind or pool relatively less sample, particularly for liquid samples, because the liquid sample has some fluidity.

In some preferred forms, as shown in fig. 10, the second sample-binding region 11a includes a curved line structure or/and a broken line structure. The curved line structure and/or the broken line structure facilitate the sample to be combined on the second sample combination area 11a, such combination area is in more contact with the surface of the sample, the sample is combined on the combination area through surface tension, attraction, viscosity and other forces, for example, the liquid sample can be combined on the second sample combination area 11a through the surface tension. In some preferred modes, the radian of the arc is greater than 0 and less than or equal to 2 pi, and when the radian is equal to 2 pi, the arc structure is a circular ring. The arc structure can be an arc or a cambered surface or a circular ring structure. The liquid has surface tension, and the liquid sample can be stably combined at the arc structure and is not easy to fall off. The broken line structure comprises a closed broken line and an open broken line, wherein the closed broken line is a polygonal structure, such as a triangle, a quadrangle, a pentagon and the like. In some preferred forms, the fold line structure further comprises a fold plane. The broken line structure all has the contained angle, the pitch arc structure has the fillet, and the surface contact of such combination region and sample is more, and liquid sample or solid-liquid mixture sample or the sample that has certain stickness combine in contained angle and fillet department easily.

In some preferred forms, the second sample binding region 11a includes a curved line structure or a polygonal line structure. In other preferred forms, the second sample binding region 11a includes both a curved line structure and a polygonal line structure. In this embodiment, the second sample combining area 11a is of a circular ring structure, as shown in fig. 10, which is beneficial to collecting a certain amount of liquid sample (or solid-liquid mixed sample or viscous sample), the sample is easily combined in the middle of the circular ring, the size of the sample left in the middle of the circular ring is affected by the size of the circular ring, so that the sample can be quantitatively collected and transferred, and the circular ring is convenient to manufacture. The sample adding head 5a with the circular ring-shaped structure can conveniently collect a certain amount of samples and can bear the samples, the samples can be stably combined at the circular ring-shaped structure, and the middle of the circular ring-shaped structure is hollowed out, so that the samples can be conveniently transferred to the detection device. The sample with certain viscosity is easily and firmly combined on the annular structure, so that the sample is favorably transferred, and the sample is not easy to leak.

In other preferred embodiments, as shown in FIG. 21, the second sample-binding region 11a has a ring-like structure, i.e., the ring may be itself notched, and the radian of the notched ring is greater than π and less than 2 π. Because the tongue sample is relatively viscous, even if the sample addition head is not a completely closed loop (e.g., a gap), it binds the same amount of sample as the closed loop.

In some preferred embodiments, as shown in fig. 10, the sample applicator 2a further comprises a second handle 6a, and the second handle 6a is connected to the sample application head directly or through other components. The second handle 6a is provided to facilitate access to the applicator 2 a. The second handle 6a and the sample adding head 5a can be fixedly connected or detachably connected, for example, the second handle 6a and the sample adding head 5a are connected by a snap structure.

In some preferred modes, the second handle 6a is made of a material with certain hardness, can keep a certain shape, and is not easy to deform, so that the sample injector 2a is convenient to take, scraping is facilitated, a sample is transferred, and the sample is added to the detection device.

In some preferred forms, the sampler 1a and the sampler 2a can be connected together and, in use, can be disconnected. As shown in fig. 11, the two are combined together to facilitate the storage, transportation and cooperation of the sampler 1a and the sampler 2a, and to avoid loss of one of them. In some preferred forms, the sampler 1a and the sampler 2a can be detachably combined, for example, by a snap connection. In other preferred embodiments, the sample can be loaded by using the sampler 1a alone and then using another device for quantitatively transferring a sample; alternatively, the sampler 1a may be used alone to collect a sample and transfer and sample the sample, or the sampler 2a may be used alone to collect a predetermined amount of sample and add the sample to the detection apparatus.

In some preferred embodiments, as shown in fig. 11, the sample application head is smaller than the sampling head, so that the sample application head can collect a certain amount of sample from the sampling head to perform sample application, and a large amount of sample is not required in the sample application.

Example 4, see figures 13 and 15.

In this embodiment, as shown in fig. 13, the first sample combining area 10a further includes a first supporting surface 12a, the sample can be combined with the first supporting surface 12a, and the first supporting surface 12a can support and bear the sample, so that the sample is not easy to separate from the first combining area 10a, and the sample is prevented from dropping or spilling elsewhere to cause contamination. In some preferred modes, the first supporting surface 12a is at least one of a plane, a concave surface and a folded surface, in this embodiment, as shown in fig. 13, the first supporting surface 12a is a plane, the plane is used as a bottom support and can support the sample, and in this embodiment, the bottom support does not completely seal the three-side ring, but leaves an opening.

In this example, the other embodiments may adopt embodiments similar to those of example 2.

Example 5, see figures 14-15.

In some preferred manners, as shown in fig. 14, the second sample combining area 11a further includes a second supporting surface 13a, the sample can be combined with the second supporting surface 13a, and the second supporting surface 13a can support and carry the sample, so that the sample is not easy to be separated from the second combining area 11a, and the sample is prevented from falling or spilling elsewhere to cause contamination. In some preferred modes, the second supporting surface 13a is at least one of a plane surface, a concave surface and a folded surface. In this embodiment, the second support surface 13a is a concave surface.

In this embodiment, the other embodiments may adopt embodiments similar to those of embodiment 3.

Example 6

The tongue coating sample in the oral cavity is collected by using the sample collecting device described in examples 2 to 5, and the helicobacter pylori detection is performed by using the detection device described in example 1, as shown in fig. 16 to 20, and specifically, the method comprises the following steps:

(1) the upper layer supporting member 1 is opened, so that the upper layer supporting member and the lower layer supporting member are flatly laid on the same plane, as shown in fig. 16 and 18;

(2) scraping a tongue coating sample on the tongue by using a sampler 1a, wherein the tongue coating sample is combined on a sampling head 3 a;

(3) using a sample applicator 2a to collect a tongue sample on a sampling head 3a, transferring the tongue sample, and applying the tongue sample to a middle reaction member 2 (including a control group and an experimental group) of the detection device or applying the sample to a corresponding position on a lower support member (the lower support member is provided with marks T and C corresponding to the experimental group and the control group, respectively, and applying the sample to an application region 10 below the marks, as shown in fig. 18);

(4) the upper layer supporting member 1 and the lower layer supporting member 3 are combined together, the sample is extruded, the sample is fully contacted with the middle layer reaction member 2, the reaction is favorably carried out, the detection is carried out rapidly, the detection result is observed within 0.5-5min, the detection result is shown in figures 17 and 19-20, and figure 19 is a negative detection result, which indicates that no helicobacter pylori exists in the tongue coating sample. FIGS. 17 and 20 are positive results indicating the presence of H.pylori in the tongue coating samples.

It is to be understood that the described embodiments are merely a few embodiments of the invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Claims

1. A detection device for detecting helicobacter pylori is characterized by comprising an upper layer supporting member, a middle layer reaction member and a lower layer supporting member, wherein the middle layer reaction member is combined with the upper layer supporting member or/and the lower layer supporting member; the substances embedded on the middle layer reaction member for performing the control test comprise a buffer solution and an acid-base indicator, and the substances embedded on the middle layer reaction member for performing the index test comprise the buffer solution, the acid-base indicator and urea.

2. The detecting device for detecting helicobacter pylori according to claim 1, wherein the substances embedded in the middle reaction member for performing the control test include a buffer solution, an acid-base indicator and a preservative, and the substances embedded in the middle reaction member for performing the index test include a buffer solution, an acid-base indicator, a preservative and urea.

3. The detecting device for detecting helicobacter pylori according to claim 1, wherein the buffer solution is at least one of a glycine buffer solution, a citric acid buffer solution, a phosphoric acid buffer solution, and a morpholine ethanesulfonic acid buffer solution having a pH of 2.0 to 7.0.

4. The detecting device for detecting helicobacter pylori according to claim 1, wherein the acid-base indicator is at least one of cresol red, phenol red and bromocresol purple.

5. The detecting device for detecting helicobacter pylori according to claim 2, wherein the preservative is at least one of sodium benzoate, sodium azide and proclin 300.

6. The detecting unit for helicobacter pylori according to claim 2, wherein the substance dissolved in the aqueous solution embedded in the middle layer reaction member used for the control test is: 0.01-0.1M citric acid buffer pH3.0-pH6.0, 0.3-30mM phenol red and 0.05-5mM sodium benzoate; substances dissolved in aqueous solution embedded on the middle layer reaction piece for index test: 0.01-0.1M citric acid buffer pH3.0-pH6.0 contains 0.3-30mM phenol red, 0.05-5mM sodium benzoate and 0.5-500mM urea.

7. The detecting device for detecting helicobacter pylori according to claim 1, wherein one end of the upper supporting member is connected to one end of the lower supporting member; the lower support or the upper support is provided with a sample adding region.

8. A method for detecting helicobacter pylori, which employs the detection device according to any one of claims 1 to 7, comprising the steps of:

9. The method for detecting helicobacter pylori according to claim 8, wherein in the step (3), the specimen is a tongue coating specimen.

10. The method of detecting helicobacter pylori according to claim 8, wherein in the step (5), the color change of the middle layer reaction member is observed within 0.5 to 5 minutes.