Detection reagent for detecting coccoid forms and spiral forms of helicobacter pylori under gastroscope and use thereof

Abstract

Provided in the present invention are a detection reagent for detecting coccoid forms and spiral forms of Helicobacter pylori under a gastroscope and the use thereof, which belong to the technical field of biomedicine. A detection reagent for detecting coccoid forms of Helicobacter pylori, which takes anthocyanin as a pH value indicator, and further comprises hydrogen peroxide. Given that the coccoid forms of Helicobacter pylori have catalase activity, the pH value is increased by means of acting on the substrate hydrogen peroxide, and then the distribution of the coccoid forms of Helicobacter pylori on gastric mucosa is displayed on the basis of a change in color of the anthocyanin.

Description

A test reagent for detecting Helicobacter pylori spherical bodies and spirochetes under gastroscopy and its application

[0001] This application claims priority to the Chinese patent application filed with the China Patent Office on December 13, 2023, with application number 202311716464.8 and invention name “A test reagent for detecting Helicobacter pylori spherical bodies and spiral bodies under gastroscopy and its application”, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The invention belongs to the technical field of biomedicine, and in particular relates to a test reagent for detecting Helicobacter pylori spirochetes and spherical bodies under gastroscopy and an application thereof. Background Art

[0003] Helicobacter pylori is a major gastric pathogen that can cause duodenal ulcers, chronic active gastritis, chronic atrophic gastritis, gastric mucosal tissue-associated lymphoma, and gastric cancer. Therefore, it is essential to quickly and easily diagnose H. pylori infection and identify the relationship between H. pylori infection and gastric diseases.

[0004] Under normal circumstances, Helicobacter pylori is active in a spiral form, causing H. pylori-related diseases. Under the influence of various adverse factors both internally and externally, H. pylori morphology mutates, transforming from a spiral to a spherical form. However, H. pylori spherical forms remain viable and highly resistant to adverse environmental conditions, allowing them to revert to a spiral form. Studies have confirmed that H. pylori spherical forms can colonize and multiply in the stomach of mice, causing gastritis. H. pylori spherical forms are a key factor in the spread of H. pylori in the human population and in the recurrence of H. pylori after treatment failure.

[0005] Current methods for detecting Helicobacter pylori include morphological testing, functional testing, and immunoassays. Morphological testing for Helicobacter pylori involves staining gastric mucosal biopsy tissue with hematoxylin and eosin, WS silver staining, Giemsa staining, and methylene blue staining to demonstrate the morphology of Helicobacter pylori, as well as bacterial culture. Functional testing for Helicobacter pylori, such as breath tests and rapid urease tests, utilizes the principle that Helicobacter pylori has urease activity that can decompose urea into ammonia (NH3) and carbon dioxide (CO2) to detect Helicobacter pylori. Immunoassays for Helicobacter pylori assess past Helicobacter pylori infection by detecting Helicobacter pylori antibodies in blood and feces.

[0006] These tests can be performed using both invasive and non-invasive methods. Non-invasive methods include breath tests, blood antibody tests, and stool antibody tests. Invasive methods require gastroscopy to obtain a biopsy of the gastric mucosa for H. pylori testing. These tests include rapid urease tests, tissue section staining, and bacterial culture.

[0007] Breath tests for Helicobacter pylori have the advantage of high sensitivity, but their disadvantages are that they can only determine whether Helicobacter pylori infection is present, but cannot determine what type of stomach disease Helicobacter pylori has caused. Helicobacter pylori antibody tests can only determine whether Helicobacter pylori has been infected in the past, but cannot determine whether Helicobacter pylori infection is currently present in the body. Rapid urease tests, tissue section staining tests, and bacterial culture have the advantage of good specificity, but have the following disadvantages: A gastric mucosal biopsy is required to detect the presence of Helicobacter pylori. Because Helicobacter pylori is distributed focally in the stomach, if the biopsy does not capture the site of Helicobacter pylori infection, a false negative result may result in a missed diagnosis.

[0008] Existing technologies for detecting Helicobacter pylori using Helicobacter pylori biological enzyme activity include the rapid urease method and the breath test method. These methods use the principle that Helicobacter pylori has urease activity that can decompose urea into ammonia (NH3) and carbon dioxide (CO2) to detect Helicobacter pylori. This mechanism can only detect the helicobacter pylori morphology, and there is currently a lack of detection reagents for the spherical body of Helicobacter pylori.

[0009] Summary of the Invention

[0010] In view of this, the purpose of the present invention is to provide a test reagent for directly detecting Helicobacter pylori spheroids under gastroscopy. Based on the fact that Helicobacter pylori spheroids have catalase activity, the pH value around Helicobacter pylori is increased by the action of the substrate hydrogen peroxide, and the purpose of detecting Helicobacter pylori spheroids on the gastric mucosa is achieved according to the color change of the pH indicator.

[0011] The invention provides a test reagent for detecting Helicobacter pylori spherical bodies, comprising anthocyanidin or anthocyanidin derivative as a pH indicator and hydrogen peroxide.

[0012] Preferably, the mass ratio of hydrogen peroxide to anthocyanin is (0.5-1.5):(0.5-1.5).

[0013] Preferably, it also includes water;

[0014] The working mass concentration of the hydrogen peroxide is 3.33% to 10%;

[0015] The working mass concentration of the anthocyanin is 3.33% to 10%.

[0016] The invention provides a test reagent for simultaneously detecting spherical bodies and spirochetes of Helicobacter pylori, comprising the test reagent and urea as a substrate for detecting spirochetes.

[0017] Preferably, the mass ratio of urea, hydrogen peroxide and anthocyanin is (1-3): (0.5-1.5): (0.5-1.5).

[0018] Preferably, the working mass concentration of the urea is 6.67% to 20%.

[0019] Preferably, the structure of the anthocyanin is as shown in Formula I:

[0020] Wherein R1 and R2 are -H, -OH or -OCH3; R3 is -H or a glycosyl group; R4 is -OH or a glycosyl group.

[0021] The present invention provides application of the test reagent in preparing a product for detecting Helicobacter pylori spherical bodies.

[0022] The present invention provides application of the test reagent in preparing a product for evaluating the therapeutic effect of Helicobacter pylori.

[0023] The present invention provides the use of the test reagent in preparing a product for screening gastric diseases associated with Helicobacter pylori infection;

[0024] The stomach-related diseases include at least one of the following: duodenal ulcer, chronic active gastritis, chronic atrophic gastritis, gastric mucosal tissue-associated lymphoma and gastric cancer.

[0025] The present invention provides a test reagent for detecting Helicobacter pylori spherical bodies, which uses anthocyanidin or anthocyanidin derivatives as a pH indicator and also includes a substrate, hydrogen peroxide. Helicobacter pylori spherical bodies have catalase activity, and catalase can decompose hydrogen peroxide into water (H2O) and oxygen (O2). Hydrogen peroxide is weakly acidic. After hydrogen peroxide decomposes into water and oxygen, the local pH value changes from weakly acidic to neutral, and the distribution of Helicobacter pylori spherical bodies can be displayed based on the change in the color of the pH indicator. It can be seen that the detection reagent provided by the present invention can accurately judge the spherical bodies of Helicobacter pylori. The detection provided by the present invention has the characteristics of good specificity and high sensitivity, and is low in cost and easy to operate. It provides great convenience for the detection of different forms of Helicobacter pylori and is suitable for clinical promotion and application.

[0026] The present invention also provides a kind of test reagent that detects Helicobacter pylori spherical body and spirochete at the same time, based on Helicobacter pylori spherical body has catalase activity, and Helicobacter pylori spirochete has urease activity, can decompose urea into ammonia (NH3) and carbon dioxide (CO2) so that the pH value of Helicobacter pylori living area is raised, after adding urea substrate, due to the existence of pH indicator, color change occurs and can show the existence and distribution of Helicobacter pylori spherical body and spirochete at the same time. Use anthocyanidin as pH indicator to add substrate hydrogen peroxide and anthocyanidin as pH indicator to add substrate urea respectively, then can show the existence and distribution of Helicobacter pylori spherical body and spirochete respectively. As can be seen, the detection reagent provided by the invention can not only accurately detect the Helicobacter pylori being infected, but also accurately judge the different forms (spirochete and spherical body) of Helicobacter pylori, the detection provided by the invention has the characteristics of good specificity and high sensitivity, and with low cost, easy to operate, for the detection of Helicobacter pylori different forms provides great convenience, is suitable for clinical application. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 shows the gastroscopy results of patients with negative Helicobacter pylori;

[0028] FIG2 shows the gastroscopy results of a patient with chronic gastritis infected with Helicobacter pylori before spraying the detection reagent;

[0029] FIG3 shows the gastroscopy results of a patient with chronic gastritis infected with Helicobacter pylori after spraying the detection reagent;

[0030] FIG4 shows the gastroscopy results of patients infected with Helicobacter pylori cocci;

[0031] Figure 5 shows the gastroscopy results of a patient with gastric mucosa-associated tissue lymphoma;

[0032] Figure 6 shows the gastroscopy results of a gastric cancer patient before spraying the test reagent;

[0033] Figure 7 shows the results of gastroscopy examination of a gastric cancer patient after spraying the detection reagent on the gastric mucosa. DETAILED DESCRIPTION

[0034] The invention provides a test reagent for detecting Helicobacter pylori spherical bodies under gastroscopy, comprising anthocyanidin or anthocyanidin derivative as a pH indicator and hydrogen peroxide as a substrate.

[0035] In the present invention, the hydrogen peroxide is a catalytic substrate of catalase produced by Helicobacter pylori spherical bodies. Hydrogen peroxide decomposes into water (H2O) and oxygen (O2) under the catalytic action of catalase. Hydrogen peroxide itself is weakly acidic. After decomposition, the local pH value changes from weakly acidic to neutral. The pH indicator shows the distribution of Helicobacter pylori spherical bodies by indicating color changes. The mass ratio of hydrogen peroxide and anthocyanin is preferably (0.5-1.5): (0.5-1.5), more preferably 1:1. When used, the detection reagent preferably also includes water. The working mass concentration of hydrogen peroxide is preferably 3.33% to 10%, more preferably 6.67%. The working mass concentration of anthocyanin is preferably 3.33% to 10%, more preferably 6.67%.

[0036] In the present invention, the anthocyanin is used as a pH indicator. The structure of the anthocyanin is shown in Formula I:

[0037] R1 and R2 are -H, -OH, or -OCH3; R3 is -H or a sugar group; and R4 is -OH or a sugar group. Anthocyanin molecules contain a highly conjugated system, acidic and basic groups, and are readily soluble in polar solvents such as water, methanol, ethanol, dilute alkalis, and dilute acids. They exhibit strong absorption in both the ultraviolet and visible regions, with maximum absorption wavelengths near 280 nm in the ultraviolet region and 500-550 nm in the visible region. The color of anthocyanins changes with pH, ​​appearing red at pH <7, purple at pH 7-8, and blue at pH >11. Repeated experiments have confirmed that anthocyanins are the most suitable pH indicators for endoscopic detection of Helicobacter pylori. Because the normal color of the gastric mucosa itself is red and it is acidic in the presence of gastric acid, anthocyanins also turn red in an acidic environment. In the presence of Helicobacter pylori, they act as substrates to increase the pH value. The color change of anthocyanins used as pH indicators can form a sharp contrast with the red color of the gastric mucosa, and can clearly show the distribution of Helicobacter pylori in the stomach. This is also one of the characteristics of this invention. The anthocyanins preferably include anthocyanins isolated from the following plants: purple cabbage, purple sweet potato, grape, blood orange, red cabbage, blueberry, eggplant, cherry, cranberry, strawberry, mulberry, hawthorn and morning glory. In an embodiment of the present invention, the anthocyanins preferably include purple cabbage pigment. Purple cabbage pigment was purchased from Qingdao Pengyuan Kanghua Natural Products Co., Ltd., China.

[0038] The present invention also provides a test reagent for simultaneously detecting Helicobacter pylori spherical bodies and Helicobacter pylori spiral bodies, comprising anthocyanin or anthocyanin derivatives as pH indicator and hydrogen peroxide and urea as substrates.

[0039] In the present invention, the urea is a catalytic substrate for urease produced by Helicobacter pylori. Under the action of urease, urea decomposes into ammonia (NH3) and carbon dioxide (CO2), which increases the pH value of the Helicobacter pylori living area. In the presence of anthocyanin, a pH indicator, a color change occurs, thereby showing the distribution of Helicobacter pylori. The mass ratio of urea, hydrogen peroxide and anthocyanin is preferably (1-3): (0.5-1.5): (0.5-1.5), more preferably 2:1:1. When used, the detection reagent preferably also includes water. The working concentration of the urea is preferably 6.67% to 20%, more preferably 13.33%.

[0040] In the present invention, when the detection reagent is used for gastroscopy, the reagent is sprayed onto the surface of the gastric mucosa using a spray tube passed through the gastroscopic biopsy channel. The spraying dose of the detection reagent is 30 to 60 ml per time.

[0041] The present invention provides the use of the test reagent for detecting Helicobacter pylori spirochetes and spherical bodies in preparing products for detecting Helicobacter pylori spirochetes and / or Helicobacter pylori spherical bodies.

[0042] In the present invention, the product preferably comprises a kit.

[0043] In the present invention, the method for detecting Helicobacter pylori spherical bodies preferably comprises the following steps:

[0044] The hydrogen peroxide is mixed with anthocyanin and water to prepare a working concentration to obtain a Helicobacter pylori spherical body detection reagent;

[0045] The Helicobacter pylori spheroid detection reagent is sprayed on the surface of the gastric mucosa to be tested from the same source as the above. When the color of the reagent on the gastric mucosa surface does not change, it indicates that the gastric mucosa is not infected with Helicobacter pylori spheroids; when the color of the reagent on the gastric mucosa surface changes from light red to blue, it indicates that the gastric mucosa is infected with Helicobacter pylori spheroids.

[0046] In the present invention, the method for detecting Helicobacter pylori spherical bodies and spiral bodies preferably comprises the following steps:

[0047] Mixing urea, hydrogen peroxide, anthocyanin and water to a working concentration to obtain a Helicobacter pylori spirochete and / or spherical body detection reagent working solution;

[0048] Spraying the detection reagent working solution on the surface of the gastric mucosa to be tested, and observing the color change of the gastric mucosa surface. When the color of the gastric mucosa surface reagent remains red and does not change, it indicates that the gastric mucosa is not infected with Helicobacter pylori spirochetes and / or Helicobacter pylori spheroids; when the color of the gastric mucosa surface reagent changes from red to blue, it indicates that the gastric mucosa is infected with Helicobacter pylori;

[0049] In order to further determine whether the infected Helicobacter pylori is in the form of a spiral or a spherical body, the detection reagent is further prepared:

[0050] Mixing urea, anthocyanin and water to prepare a working concentration to obtain a Helicobacter pylori spirochete detection reagent;

[0051] Spraying the Helicobacter pylori detection reagent on the surface of the gastric mucosa to be tested from the same source as the above, when the color of the reagent on the gastric mucosa surface does not change, it indicates that the gastric mucosa is not infected with Helicobacter pylori; when the color of the reagent on the gastric mucosa surface changes from red to blue, it indicates that the gastric mucosa is infected with Helicobacter pylori;

[0052] The hydrogen peroxide is mixed with anthocyanin and water to prepare a working concentration to obtain a Helicobacter pylori spherical body detection reagent;

[0053] The Helicobacter pylori spheroid detection reagent is sprayed on the surface of the gastric mucosa to be tested from the same source as the above. When the color of the reagent on the gastric mucosa surface does not change, it indicates that the gastric mucosa is not infected with Helicobacter pylori spheroids; when the color of the reagent on the gastric mucosa surface changes from light red to blue, it indicates that the gastric mucosa is infected with Helicobacter pylori spheroids.

[0054] In the present invention, the detection reagent can perform real-time detection of Helicobacter pylori infection during gastroscopy observation, and can also perform other related tests on biopsy tissue of gastric mucosa at the discolored area. It has great operational convenience and is safe and has no side effects.

[0055] Since Helicobacter pylori spherical bodies are an important factor affecting the spread of Helicobacter pylori in the population and are also an important factor in the recurrence of Helicobacter pylori after treatment. Therefore, the present invention provides the use of the test reagent for simultaneously detecting Helicobacter pylori spirals and spherical bodies in the preparation of products for evaluating the treatment effect of Helicobacter pylori and / or the transmissibility of Helicobacter pylori.

[0056] In the present invention, the above-mentioned detection method is used to evaluate the therapeutic effect of Helicobacter pylori. Specifically, the Helicobacter pylori spirochete detection reagent working solution is sprayed on the surface of the gastric mucosa to be tested. A negative test result appears (the color of the reagent on the surface of the gastric mucosa is red and does not change). The Helicobacter pylori spherical body detection reagent is sprayed on the gastric mucosa of the same source for detection again. When the positive test result appears (the color of the reagent on the surface of the gastric mucosa changes from red to blue-purple), it indicates that the Helicobacter pylori in the gastric mucosa of the patient to be tested has undergone spherical transformation, and the Helicobacter pylori has not been eradicated and there is a possibility of recurrence.

[0057] Since Helicobacter pylori is an important pathogenic bacterium of the stomach and can cause the occurrence and development of various gastric diseases, the present invention provides the use of the test reagent for simultaneously detecting Helicobacter pylori spirochetes and spherical bodies in the preparation of products for screening gastric diseases related to Helicobacter pylori infection.

[0058] In the present invention, the gastric disease associated with Helicobacter pylori infection preferably includes at least one of the following: duodenal ulcer, chronic active gastritis, chronic atrophic gastritis, gastric mucosal tissue-associated lymphoma, and gastric cancer. In the embodiments of the present invention, by screening the causes of gastric diseases associated with Helicobacter pylori infection, the infection status of Helicobacter pylori in various gastric diseases is clarified, thereby providing guidance for subsequent treatment of gastric diseases.

[0059] The following describes in detail a test reagent for detecting Helicobacter pylori spirochetes and spherical bodies under gastroscopy and its application in conjunction with the embodiments provided by the present invention, but they should not be construed as limiting the scope of protection of the present invention.

[0060] Example 1

[0061] A test reagent for simultaneously detecting Helicobacter pylori spirochetes and spherical bodies and its use method

[0062] 1. Reagent composition: solid urea, solid hydrogen peroxide and solid purple cabbage pigment;

[0063] 2. Preparation of reagents: Weigh 4g, 2g, and 2g of solid urea, solid hydrogen peroxide, and solid purple cabbage pigment respectively, mix them, and dissolve them in 30ml of water to obtain the detection reagent working solution.

[0064] 3. How to use the detection reagent

[0065] During gastroscopy, the patient's gastric fluid is aspirated through the gastroscopic biopsy forceps channel, 30 ml of the test reagent working solution is drawn into a syringe, and the spray tube is inserted through the gastroscopic biopsy forceps channel. The test reagent working solution is sprayed onto the surface of the gastric mucosa, and the color change of the reagent on the surface of the gastric mucosa is observed through the gastroscope:

[0066] Helicobacter pylori spirochetes possess urease activity, which breaks down urea into ammonia (NH3) and carbon dioxide (CO2), raising the pH in areas where H. pylori live. Helicobacter pylori spherical bodies do not produce urease, but instead produce catalase, which breaks down hydrogen peroxide into water (H2O) and oxygen (O2): 2H2O2 = 2H2O + O2. Hydrogen peroxide is weakly acidic, and after decomposing into water and oxygen, the local pH changes from weakly acidic to neutral. Purple cabbage pigment, used as a pH indicator, displays red in acidic conditions, blue in neutral conditions, and green in alkaline conditions. Its color change indicates the distribution of Helicobacter pylori.

[0067] For those without H. pylori infection, the color of the reagent on the gastric mucosa remains red, consistent with the color of the gastric mucosa. For those with H. pylori spirochetes or coccidia, the color of the reagent on the gastric mucosa changes from red to blue.

[0068] In order to determine the morphology of Helicobacter pylori, Helicobacter pylori spirochete detection reagent (4g solid urea, 2g solid purple cabbage pigment, dissolved in 30ml water) and Helicobacter pylori spherical body detection reagent (2g solid hydrogen peroxide, 2g solid purple cabbage pigment, dissolved in 30ml water) were prepared respectively, and the gastric mucosa was sprayed again for detection. According to the results of each detection reagent, it can be determined whether the Helicobacter pylori infected by the patient is a spirochete or a spherical body of Helicobacter pylori.

[0069] Example 2

[0070] Experiments using detection reagents to test patients with different gastric diseases

[0071] The relationship between Helicobacter pylori infection and chronic gastritis, gastric cancer, and gastric mucosal tissue-associated lymphoma was detected. While performing gastroscopy on patients with chronic gastritis, gastric cancer, and gastric lymphoma, Helicobacter pylori spirochetes and spherical bodies were detected using the detection reagent of Example 1. The detection steps were the same as those in Example 1.

[0072] Figure 1 shows the results of gastroscopy for patients with negative Helicobacter pylori. As can be seen from Figure 1, the surface of the gastric mucosa is smooth and flat, and there is no color change after spraying the detection reagent, indicating that the patient has neither Helicobacter pylori spirochete infection nor Helicobacter pylori cocci infection.

[0073] Figure 2 shows the results of a gastroscopy of a patient with chronic gastritis infected with Helicobacter pylori before spraying with the test reagent. As can be seen in Figure 2, the gastric mucosal surface is rough with scattered erosions. Figure 3 shows the results of a gastroscopy after spraying with the test reagent. As can be seen in Figure 3, some areas of the gastric mucosal surface exhibit a noticeable bluish-purple discoloration, indicating that the patient is currently infected with Helicobacter pylori.

[0074] Figure 4 shows the gastroscopy results of a patient infected with Helicobacter pylori cocci. The patient underwent a follow-up gastroscopy after a negative breath test for Helicobacter pylori infection following medication. As can be seen in Figure 4, the patient's gastric mucosa was swollen, and the color remained unchanged after spraying with the Helicobacter pylori spirochete detection reagent. However, after spraying with the Helicobacter pylori cocci detection reagent, the gastric mucosa took on a light blue hue, indicating that the patient currently has Helicobacter pylori cocci infection and is at risk of recurrence.

[0075] Figure 5 shows the gastroscopy results of a patient with gastric mucosa-associated lymphoma. As can be seen in Figure 5, the patient's gastric mucosa is swollen and the mucosal folds have disappeared, but the mucosal surface is intact. After spraying the test reagent, a clear blue-purple color change was observed on the gastric mucosa, indicating that the patient was positive for Helicobacter pylori.

[0076] Figure 6 shows the gastroscopy results of a gastric cancer patient before spraying with the test reagent. As can be seen in Figure 6, there is a distinct crater-shaped ulcerated mass in the lesser curvature of the gastric antrum, with surrounding mucosa and a central ulcer. After spraying with the test reagent, the periphery of the mass has a light blue hue. Figure 7 shows a clear blue-purple linear color change in the gastric mucosa of the lesser curvature of the gastric body, indicating Helicobacter pylori infection in this patient.

[0077] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A test reagent for detecting Helicobacter pylori spheroids, characterized in that, It includes the pH indicator anthocyanin or anthocyanin derivative and hydrogen peroxide.

2. The test reagent for detecting Helicobacter pylori spheroids according to claim 1, characterized in that, The mass ratio of the hydrogen peroxide to the anthocyanin is (0.5 - 1.5):(0.5 - 1.5).

3. The test reagent according to claim 1 or 2, characterized in that, It also includes water; The working mass concentration of the hydrogen peroxide is 3.33% - 10%; The working mass concentration of the anthocyanin is 3.33% - 10%.

4. A test reagent for simultaneously detecting Helicobacter pylori spheroids and spirochetes, characterized in that, It includes the test reagent according to any one of claims 1 - 3 and the substrate urea for detecting Helicobacter pylori.

5. The test reagent according to claim 4, characterized in that, The mass ratio of the urea, the hydrogen peroxide and the anthocyanin is (1 - 3):(0.5 - 1.5):(0.5 - 1.5).

6. The test reagent according to claim 4 or 5, characterized in that, The working mass concentration of the urea is 6.67% - 20%.

7. The test reagent for detecting Helicobacter pylori spheroids according to claim 1 or 2 or the test reagent for simultaneously detecting Helicobacter pylori spheroids and spirochetes according to claim 4 or 5, characterized in that, The structure of the anthocyanin is shown in Formula I: Wherein R1 and R2 are -H, -OH or -OCH3; R3 is -H or glycosyl; R4 is -OH or glycosyl.

8. Use of the test reagent for detecting Helicobacter pylori spheroids according to claim 1 or 2 or the test reagent for simultaneously detecting Helicobacter pylori spheroids and spirochetes according to any one of claims 4 to 7 in the preparation of a product for detecting Helicobacter pylori spheroids.

9. Use of the test reagent for detecting Helicobacter pylori spheroids according to claim 1 or 2 or the test reagent for simultaneously detecting Helicobacter pylori spheroids and spirochetes according to any one of claims 4 to 7 in the detection of Helicobacter pylori spheroids.

10. Use of the test reagent according to any one of claims 4 to 7 in the preparation of a product for detecting Helicobacter pylori spheroids and spirochetes.

11. Use of the test reagent according to any one of claims 4 to 7 in the simultaneous detection of Helicobacter pylori spheroids and Helicobacter pylori spirochetes.

10. Use of the test reagent for detecting Helicobacter pylori spheroids according to claim 1 or 2 or the test reagent for simultaneously detecting Helicobacter pylori spheroids and spirochetes according to any one of claims 4 to 7 in the preparation of a product for evaluating the therapeutic effect of Helicobacter pylori.

11. Use of the test reagent for detecting Helicobacter pylori spheroids according to claim 1 or 2 or the test reagent for simultaneously detecting Helicobacter pylori spheroids and spirochetes according to any one of claims 4 to 7 in the evaluation of the therapeutic effect of Helicobacter pylori.

12. Use of the test reagent for detecting Helicobacter pylori spheroids according to claim 1 or 2, or the test reagent for simultaneously detecting Helicobacter pylori spheroids and spirochetes according to any one of claims 4 to 7, in the preparation of a product for screening gastric diseases related to Helicobacter pylori infection; The gastric-related diseases include at least one of the following: duodenal ulcer, chronic active gastritis, chronic atrophic gastritis, gastric mucosa-associated lymphoma, and gastric cancer.

13. Use of the test reagent for detecting Helicobacter pylori spheroids according to claim 1 or 2, or the test reagent for simultaneously detecting Helicobacter pylori spheroids and spirochetes according to any one of claims 4 to 7, in screening gastric diseases related to Helicobacter pylori infection; The gastric-related diseases include at least one of the following: duodenal ulcer, chronic active gastritis, chronic atrophic gastritis, gastric mucosa-associated lymphoma, and gastric cancer.

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