Kit and probiotic product for predicting, diagnosing, preventing and treating gastric disease occurrence risk of helicobacter pylori infected person
By screening microorganisms specific to Helicobacter pylori infected people to make test kits and microorganisms specific to healthy people to make probiotic products, the problem of poor treatment of gastric diseases caused by Helicobacter pylori infection has been solved, efficient risk prediction and treatment effects have been achieved, and the increased drug resistance caused by the abuse of antibiotics has been avoided.
Patent Information
- Application Number
- CN202410362767.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-03-20
- Publication Date
- 2025-09-23
AI Technical Summary
In the existing technology, the treatment effect of gastric diseases caused by Helicobacter pylori infection is poor, especially due to antibiotic resistance and changes in the structure of microbial flora, which make traditional treatment options inefficient and have the risk of enhanced drug resistance.
By screening the unique and enriched microorganisms in Helicobacter pylori-infected people, a test kit is made for disease risk prediction and diagnosis. Probiotic products are made using microorganisms unique to healthy people to prevent and treat gastric diseases and avoid the increase in drug resistance caused by the abuse of antibiotics.
It achieves high accuracy in predicting and diagnosing the risk of gastric diseases in patients infected with Helicobacter pylori, while providing effective prevention and treatment options without enhancing drug resistance, thus avoiding the side effects of antibiotics.
Smart Images
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Abstract
Description
Technical Field
[0001] The present invention relates to the field of microbial technology, in particular to a kit and probiotic product for predicting, diagnosing, preventing and treating gastric diseases in patients infected with Helicobacter pylori. (1) A class of microorganisms that are unique to and enriched in patients with gastric diseases in Helicobacter pylori-infected patients are screened for use in preparing a kit for predicting and diagnosing the risk of gastric diseases in patients infected with Helicobacter pylori; (2) A class of microorganisms that are unique to or enriched in healthy people are screened for use in preparing a probiotic product for preventing and treating gastric diseases. The kit product and probiotic product of this technology can be used for predicting and diagnosing the risk of gastric diseases in patients infected with Helicobacter pylori, and the probiotic product can prevent and treat related gastric diseases under the premise of avoiding the development or enhancement of drug resistance in Helicobacter pylori. Background Art
[0002] Helicobacter pylori (H. pylori) is a microaerophilic, Gram-negative, spiral-shaped bacterium. It has attracted widespread attention due to its widespread infection (over 50%) and potential to cause serious gastric diseases (such as gastric cancer and gastric adenocarcinoma). This bacterium is highly colonized in the gastric mucosa, making spontaneous clearance difficult. Its metabolic factors can induce and activate inflammatory responses, destroy gastric mucosal tissue, and activate cancer-related pathways (such as the Wnt signaling pathway), leading to diseases such as gastritis, atrophic gastritis, and gastric cancer, posing a serious threat to human health. The gastric microbiome primarily comprises five phyla: Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria, and Fusobacteria. H. pylori is the only member of the genus Helicobacter. Studies have shown that after infection with H. pylori, its relative abundance significantly dominates, while the relative abundance of other microorganisms decreases. Secondly, there were significant differences in the diversity and composition of the gastric microbiome across disease stages (healthy individuals, gastritis, atrophic gastritis, intestinal metaplasia, and gastric cancer). There were also significant differences in the relative abundance of gastric microorganisms between H. pylori-negative and H. pylori-positive individuals at different disease stages. This suggests that H. pylori infection has affected and altered the composition and structure of the gastric microbiome. It is possible that H. pylori infection is responsible for the disturbance in the gastric microbiome, which in turn leads to gastric disease.
[0003] First-line antibiotic treatment is currently a common strategy for eliminating Helicobacter pylori infection. It has evolved from traditional triple therapy to the latest bismuth-containing quadruple therapy. However, due to the development of bacterial resistance to commonly prescribed antibiotics and the adverse reactions associated with antibiotic treatment, the percentage of treatment failure continues to rise. Clinically used antibiotics for Helicobacter pylori eradication include nitroimidazoles, macrolides, quinolones, tetracyclines, and furazolidone. Helicobacter pylori has a high resistance rate to clarithromycin, metronidazole, and levofloxacin, and its resistance is increasing with the overuse and irrational use of antibiotics. Furthermore, due to regional differences in antibiotic types and usage habits, resistance rates vary across regions.
[0004] In summary, Helicobacter pylori is highly correlated with the development and progression of gastric diseases, and its eradication treatment primarily relies on antibiotics. However, H. pylori currently possesses a high degree of drug resistance. Even if treatment strategies could be developed that circumvent H. pylori's drug resistance and utilize non-drug-resistant antibiotics, long-term use of such treatments still carries a risk of enhancing drug resistance in H. pylori. Furthermore, H. pylori infection significantly alters the structure of the gastric microbiome. Therefore, if H. pylori's strong association with other bacterial species could be exploited to enhance H. pylori's "enemy" species, thereby inhibiting or neutralizing H. pylori's toxins, this would truly achieve the effect of "treating bacteria with bacteria." This probiotic therapy would effectively mitigate and prevent the development of drug resistance caused by overuse of antibiotics. Summary of the Invention
[0005] The objects of the present invention are: The objectives of the present invention are to (1) prepare a kit product using microorganisms that are unique or enriched in patients with gastric diseases (chronic gastritis, superficial gastritis, atrophic gastritis, intestinal metaplasia, gastric cancer) infected with Helicobacter pylori, for use in predicting and diagnosing the risk of gastric diseases, and (2) prepare a probiotic product using microorganisms that are unique or enriched in healthy people infected with Helicobacter pylori, for use in preventing and treating gastric diseases.
[0006] To achieve the above object, the technical solution adopted by the present invention is: (1) The present invention studies the gastric microbial flora of healthy people and patients with five types of gastric diseases among Helicobacter pylori infected people. The richness-distribution index of each microorganism in the sample is calculated by combining the method proposed by the inventor with the random test, and a strict statistical test (random test) is performed to eliminate the influence of individual differences in samples on the results. The unique microorganisms and enriched microorganisms of healthy people and patients with gastric diseases among Helicobacter pylori infected people are screened (p < 0.05, after FDR correction). Claim 1 lists the unique microorganisms and enriched microorganisms of each type of gastric disease, which can be used as targets for predicting the risk of gastric disease and diagnosing diseases. Claim 1 also lists the unique microorganisms and enriched microorganisms of healthy tissues, which can be used as potential probiotics for preventing and treating gastric diseases. The inventors used rigorous statistical tests and performed false-positive corrections, and the microorganisms that were unique and enriched in patients with gastric diseases among Helicobacter pylori-infected people screened out had high accuracy in detecting the risk of gastric diseases and diagnosing the diseases; the microorganisms that were unique and enriched in healthy people screened out are potential probiotics for the prevention and treatment of related gastric diseases, and can avoid the development and enhancement of Helicobacter pylori resistance. (2) The gastric microbial flora samples of the subjects are collected by gastroscopy and other means, and the test tissues are tested using products such as kits containing specific microorganisms and enriched microorganisms of patients with gastric diseases infected with Helicobacter pylori. If the test result is positive, it means that the subject may have or already has gastric disease. (3) The reagents required for the implementation of the application of the present invention include at least one of a metagenomic sequencing reagent, a 16S rRNA amplicon sequencing reagent, a droplet digital PCR detection reagent, and a qPCR quantitative detection reagent. The product of the present invention will detect the entire sequence of unique species and enriched species or any portion of the entire sequence or its barcode marker sequence or the expression products and metabolites of the sequence. The unique microorganisms and enriched microorganisms screened out for patients with gastric diseases are used as detection targets, and at least one of the above methods is used to determine whether one or more types of microbial markers are present in gastric diseases. (4) The focus of the present invention is to screen out the microorganisms that are unique to various types of gastric disease patients and the enriched microorganisms in patients infected with Helicobacter pylori, and the microorganisms that are unique to and enriched in healthy people relative to gastric disease patients, and use the microorganisms that are unique to and enriched in gastric disease patients as targets to predict the risk of gastric disease and diagnose. Corresponding kit products are prepared to detect whether one or more types of microbial markers that are unique to gastric disease patients are present in the tissue to be tested. Probiotic products are prepared using the microorganisms that are unique to healthy people relative to gastric disease patients as the core means of gastric disease prevention and treatment. (5) The present invention uses specific microbial markers to detect the risk of gastric disease. It is not limited to the measurement methods and means of microbial markers, as long as it can detect whether one or several types of microbial markers specific to gastric disease patients are present in the tissue to be tested. (6) The present invention uses microorganisms unique to healthy people and enriched microorganisms in Helicobacter pylori-infected people as the core method for preventing and treating gastric diseases. It is not limited to the method of preventing and treating gastric diseases, as long as one or several species of microorganisms unique to healthy tissues and enriched microorganisms are used as the core means of treatment. (7) The microorganisms unique to healthy tissues and the enriched microorganisms screened by the present invention can be used as a means of preventing and treating gastric diseases by inhibiting Helicobacter pylori toxins or by any other method. This includes any drugs, health products, probiotics, devices, etc. that use this as a core method for treating gastric diseases.
[0007] The effects of the present invention are: The effects of the present invention are: (1) a kit for predicting and diagnosing the risk of developing a disease is invented using microorganisms that are unique and enriched in patients with gastric diseases (chronic gastritis, superficial gastritis, atrophic gastritis, intestinal metaplasia, and gastric cancer) infected with Helicobacter pylori; and (2) a probiotic product for preventing and treating gastric diseases is invented using microorganisms that are unique and enriched in healthy people infected with Helicobacter pylori. The risk of developing, diagnosing, preventing, and treating related gastric diseases can be predicted, diagnosed, and treated without developing or enhancing drug resistance in Helicobacter pylori. DETAILED DESCRIPTION
[0008] In order to better illustrate the technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with a specific analysis process.
[0009] The inventors collected gastric microbial flora samples from a database of patients with five gastric diseases and healthy individuals (infected with H. pylori but without gastric diseases). The species screening process was demonstrated using gastric cancer data as an example. The specificity index for each species was calculated using the richness-distribution index: D i h Indicates the extent of distribution of species i, P i hRepresents the relative abundance of the i-th species in the microbial community across all samples. The richness-distribution index combines species distribution frequency and relative richness to assess species. This method overcomes the shortcomings of previous assessment methods that only consider relative abundance or distribution frequency. The new method screens more reliable species.
[0010] A random test was conducted on the abundance and distribution index of each microorganism in gastric cancer patients and healthy people among Helicobacter pylori-infected individuals to eliminate the influence of individual and random factors on the results. The microorganisms unique to gastric cancer patients and healthy people were screened for enrichment (p < 0.05, after FDR correction). Claim 1 lists the microorganisms unique to gastric cancer patients and enrichment, as well as the microorganisms unique to healthy tissue relative to patients. The random test was conducted by the following steps: S1: Calculate the richness-distribution index of each species in healthy people and gastric cancer patients respectively, and calculate the absolute value Δ of the difference in richness-distribution index of each species in healthy people and gastric cancer patients; S2: Mix samples from healthy people and gastric cancer patients, and randomly divide them into two groups A' and B'. Calculate the richness-distribution index of each species in the two groups A' and B'. Calculate the absolute value Δ' of the difference in the richness-distribution index of each species in A' and B'. S3: Repeat step S2 1000 times to obtain 1000 Δ's; S4: Calculate the p-value of each species according to the formula p = n / 1000, where n represents the number of 1000 Δ's greater than Δ in S3; S5: Species unique to healthy populations must meet the following conditions: p < 0.05, the richness-distribution index in healthy populations is not equal to 0, and the richness-distribution index in gastric cancer patients is equal to 0; S6: Species unique to gastric cancer patients must meet the following conditions: p < 0.05, the richness-distribution index in gastric cancer patients is not equal to 0, and the richness-distribution index in healthy people is equal to 0; S7: Dominant species in healthy individuals (species with significantly increased richness-distribution indices compared to gastric cancer patients) must meet the following conditions: p < 0.05, the richness-distribution indices in both healthy individuals and gastric cancer patients are not equal to 0, and the richness-distribution indices in healthy individuals are greater than those in gastric cancer patients; S8: The dominant species in gastric cancer patients (species with significantly increased abundance-distribution index compared to healthy tissues) must meet the following conditions: p < 0.05, the abundance-distribution index in both healthy people and gastric cancer patients is not equal to 0, and the abundance-distribution index in gastric cancer patients is greater than that in healthy tissues;
[0011] Following the above steps, we screened for species unique to and enriched in gastric cancer patients, and species unique to and enriched in healthy tissues. Species unique to and enriched in healthy individuals can be developed into probiotic products for the prevention and treatment of gastric cancer, while species unique to and enriched in gastric cancer patients can be used to develop kits for gastric cancer risk prediction and diagnosis. The species list is as follows:
Claims
1. A kit and probiotic product for predicting, diagnosing, preventing and treating the risk of gastric diseases in patients infected with Helicobacter pylori. One type of microorganisms, unique to or enriched in patients with each type of gastric disease infected with Helicobacter pylori, are used to develop kits for predicting and diagnosing the risk of gastric diseases. Another type of microorganisms, unique to or enriched in healthy people infected with Helicobacter pylori, are used to develop probiotic products for preventing and treating gastric diseases, including chronic gastritis, superficial gastritis, atrophic gastritis, intestinal metaplasia, and gastric cancer. The specific and enriched microorganisms in patients with each type of gastric disease in patients infected with Helicobacter pylori are: The specific and enriched microorganisms in healthy individuals relative to each gastric disease in patients with Helicobacter pylori infection are:
2. The product according to claim 1, characterized in that A kit for risk prediction and diagnosis of any one or more of five gastric diseases and a probiotic product for prevention and treatment.
3. The product according to claim 1, characterized in that The microorganisms are one or a combination of several or all of the microorganisms.
4. The product according to claim 1, characterized in that A product that uses the entire gene sequence or any part of the entire sequence or characteristic barcode sequence or sequence expression products and metabolites of a type of microorganism that is unique and enriched in patients with gastric diseases among Helicobacter pylori infected patients as detection targets.
5. The product according to claim 1, characterized in that The application of a type of microorganisms that are unique and enriched in patients with gastric diseases among Helicobacter pylori infected people in the preparation of gastric disease risk prediction and diagnosis kit products and sub-products.
6. The product according to claim 1, characterized in that The application of a type of microorganisms that are unique and enriched in patients with gastric diseases among Helicobacter pylori infected people in the preparation of software, hardware and firmware products for gastric disease risk prediction and diagnosis kit products.
7. The product according to claim 1, characterized in that Foods, medicines, health products and derivative products that use one or a combination of several or all of the microorganisms that are unique or enriched in healthy people infected with Helicobacter pylori as probiotics.
8. The product according to claim 1, characterized in that Foods, medicines, health products and derivatives that use as ingredients the entire gene sequence or any part of the entire sequence or the sequence expression products and metabolites of microorganisms that are unique or enriched in healthy people infected with Helicobacter pylori.
9. The product according to claim 1, characterized in that Software and hardware products that use one or more bacterial consortia of microbial species that are unique or enriched in healthy people infected with Helicobacter pylori as a means of prevention and treatment.
10. The product according to claim 1, characterized in that Software and hardware products that use the entire gene sequence or any part of the entire sequence or the sequence expression products and metabolites of microbial species that are unique or enriched in healthy people infected with Helicobacter pylori as a means of prevention and treatment.