CN-121991822-A - Combined preparation for promoting helicobacter pylori tricarboxylic acid to circularly synthesize ATP, and preparation method and application thereof

Abstract

A combined preparation for promoting helicobacter pylori tricarboxylic acid to circularly synthesize ATP, and its preparation method and application are provided. The preparation is based on brain heart infusion medium and 10wt.% calf serum, by adding 1mM L-cysteine, 2mM L-glutamine and 5 μg/mL heme as key nutritional supplements and adjusting the pH of the medium to 7.35. The application of the combined preparation obviously improves the in-vitro culture efficiency of helicobacter pylori, can rapidly increase bacteria in a short time, has low cost, and is suitable for isolated culture and rapid passage of helicobacter pylori in scientific research and clinical samples.

Inventors

• HUANG YANQIANG
• SHEN YALING
• ZHAO HAOLIANG
• Luo Jiazi
• HUANG LINGLI

Assignees

• 右江民族医学院

Dates

Publication Date

20260508

Application Date

20251218

Claims (9)

1. 1. A combined preparation for promoting the circulating synthesis of ATP by helicobacter pylori tricarboxylic acid is characterized in that the main components comprise brain heart infusion culture medium and calf serum, and the pH value of the combined preparation is 7.35.
2. 2. The combination preparation of claim 1, further comprising 1mM L-cysteine, 2mM L-glutamine, 5 μg/mL heme.
3. 3. The combination preparation according to claim 1, wherein the brain heart infusion medium is used in an amount of 13.33g per 400mL final medium and the calf serum is used in an amount of 40mL.
4. 4. A process for preparing a combined preparation for promoting cyclic synthesis of ATP by helicobacter pylori tricarboxylic acid as defined in any one of claims 1 to 3, which comprises the steps of (a) mixing a brain heart infusion medium with water, sterilizing and cooling to about 50 ℃, (b) adding calf serum, L-cysteine, L-glutamine and heme to the medium obtained in the step (a), mixing uniformly, and (C) adjusting the pH value of the mixture obtained in the step (b) to 7.35.
5. 5. The method of claim 4, wherein in step (a), the sterilization is autoclaving.
6. 6. The method according to claim 5, wherein in the step (b), the components are added in the order of adding calf serum followed by L-cysteine, L-glutamine and heme.
7. 7. A culture medium for isolated culture of helicobacter pylori, characterized in that it comprises a combined preparation as defined in any one of claims 1 to 4.
8. 8. The culture medium according to claim 7, which is a liquid culture medium or a solid culture medium.
9. 9. Use of a combined preparation according to any one of claims 1 to 4 or a culture medium according to any one of claims 7 to 8 for the preparation of a product for the isolation, cultivation or proliferation of helicobacter pylori.

Description

Combined preparation for promoting helicobacter pylori tricarboxylic acid to circularly synthesize ATP, and preparation method and application thereof Technical Field The invention belongs to the field of medicine and health, and in particular relates to a combined preparation for promoting cyclic synthesis of ATP (adenosine triphosphate) by helicobacter pylori tricarboxylic acid, and a preparation method and application thereof. Background Helicobacter pylori (Helicobacter pylori, h. Pylori) is a microaerophilic gram-negative bacillus colonized on human gastric mucosa, and its infection is closely related to the occurrence and development of chronic gastritis, peptic ulcer, gastric mucosa-associated lymphoid tissue lymphoma, gastric cancer and other various digestive tract diseases, and constitutes a great burden on global public health. Therefore, the helicobacter pylori is accurately and efficiently detected and isolated and cultured, and has important significance for clinical diagnosis of diseases, epidemiological investigation, antibiotic susceptibility test and pathogenic mechanism research. At present, isolated culture of helicobacter pylori in vitro is still one of the "gold standards" for diagnosis and provides the necessary viable bacterial material for subsequent studies. Traditional culture methods rely mainly on nutrient-rich solid media such as Columbia blood agar, brain heart infusion agar, etc., typically supplemented with animal blood (e.g., sheep blood, horse blood) or serum and provided a microaerophilic environment (typically 85% N 2、10% CO2、5% O2). However, helicobacter pylori is extremely demanding on growth conditions, has slow growth and low positive rate of isolated culture, and is always a main technical problem which afflicts clinical and scientific research work. In particular, helicobacter pylori grows slowly on conventional media, typically requiring 3 to 5 days of culture, and even longer to form macroscopic colonies, which greatly prolongs the diagnostic period, and is difficult to meet the clinical rapid diagnosis requirements. More importantly, the slow growth rate may cause the living bacteria to die or enter a non-culturable state during the cultivation process due to inadaptation to the in vitro environment, so that many truly existing infections cannot be detected by a cultivation method, and false negative results and the positive detection rate are often not ideal. In addition, the success rate of primary separation of helicobacter pylori from clinical gastric mucosa biopsy specimens is lower, because thalli after gastroscopic material drawing, transportation and pretreatment may be in a sub-injury state and is more sensitive to a culture environment. Although attempts have been made in the prior art to improve the growth of H.pylori by adding different nutrients to the basal medium, these improvements often suffer from complex ingredients, high costs, cumbersome preparation or unstable effects. For example, some modified media may focus on providing a rich carbon and nitrogen source, but ignore the special metabolic demands of H.pylori for amino acids, reducing agents, and iron sources in microaerophilic environments, which are critical for their rapid initiation of proliferation, breakthrough of growth lag phase. Therefore, a combined preparation which has definite components, simple and convenient preparation and reasonable cost and can obviously promote the rapid growth and high positive rate separation of helicobacter pylori is developed, and the combined preparation has urgent need and important application value for improving the relevant clinical detection level and basic research efficiency of helicobacter pylori. Disclosure of Invention The invention provides a combined preparation for promoting the cyclic synthesis of ATP by helicobacter pylori tricarboxylic acid, a preparation method and application thereof, which can promote the rapid proliferation of helicobacter pylori in any isolated culture product, has simple and convenient operation and low cost, and is suitable for wide popularization and application in scientific research and clinical fields. The technical scheme is that the combined preparation for promoting the circulation of helicobacter pylori tricarboxylic acid into ATP mainly comprises brain heart infusion culture medium and calf serum, and the pH value of the combined preparation is 7.35. The above combination preparation further comprises 1mM L-cysteine, 2mM L-glutamine, 5. Mu.g/mL heme. The brain heart infusion medium is used in an amount of 13.33g per 400mL of final medium, and the calf serum is used in an amount of 40mL. The preparation method of the combined preparation for promoting the cyclic synthesis of ATP by helicobacter pylori tricarboxylic acid comprises the following steps of (a) mixing a brain heart infusion culture medium with water, sterilizing and cooling to about 50 ℃, b) adding calf serum, L-cysteine, L-glutamine and he