AU2021105934A4 - Application of Lactobacillus Acidophilus LA-10A in Prevention and Treatment of Helicobacter Pylori Infection - Google Patents

Application of Lactobacillus Acidophilus LA-10A in Prevention and Treatment of Helicobacter Pylori Infection Download PDF

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AU2021105934A4
AU2021105934A4 AU2021105934A AU2021105934A AU2021105934A4 AU 2021105934 A4 AU2021105934 A4 AU 2021105934A4 AU 2021105934 A AU2021105934 A AU 2021105934A AU 2021105934 A AU2021105934 A AU 2021105934A AU 2021105934 A4 AU2021105934 A4 AU 2021105934A4
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helicobacterpylori
lactobacillus acidophilus
cells
strains
screening
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Xin Ma
Xueping Yu
Yang Yu
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Thankcome Biological Science and Technology Suzhou Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/18—Testing for antimicrobial activity of a material
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/113—Acidophilus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K2035/11—Medicinal preparations comprising living procariotic cells
    • A61K2035/115—Probiotics
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus
    • C12R2001/23—Lactobacillus acidophilus
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
    • G01N2333/205—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Campylobacter (G)

Abstract

According to the invention, Lactobacillus acidophilus LA-iOA capable of inhibiting Helicobacterpylori is screened out to reduce or prevent Helicobacterpylori infection, and the strain can be used for preparing functional foods or pharmaceutical compositions.

Description

Application of Lactobacillus Acidophilus LA-10A in Prevention and Treatment of
Helicobacter Pylori Infection
TECHNICAL FIELD
The invention belongs to the field of biotechnology, and particularly relates to the application
of Lactobacillus acidophilus LA-10A in prevention and treatment of Helicobacter pylori
infection.
BACKGROUND
Helicobacterpylori(HP) is the main cause of chronic gastritis, duodenitis and gastric ulcer, and
it is also a major risk factor for gastric cancer and gastric lymphoma. More than 50% of people
in the world are infected with HP, but only about 15% of people will cause diseases. The causes
of this result are not only related to the genetic susceptibility and environmental factors of the
host itself, but also related to the virulence of the infected strains. With the development of
modem biomolecular technology, people have found HP strains with different virulence, but the
eradication rates of HP strains with different virulence are different by traditional triple or
quadruple therapy, and 10%-25% of patients may fail to eradicate for the first time. Failure in
eradication not only has a great impact on patients' lives and psychology, but also easily
produces drug-resistant bacteria. At the same time, for many HP-positive patients infected with
low virulence strains, the side effects caused by antibiotic treatment may exceed the effects
caused by infection with low virulence strains. For patients infected with HP strains, it is of
great practical significance to provide a new treatment which has almost no side effects and can
control or eradicate HP for a long time. The microbiota therapy of "treating bacteria with
bacteria" provides a new idea. Many animal experiments have shown that HP can colonize the
stomach of sterile mice and cause inflammation of gastric mucosa, while in mice fed probiotics, the colonization of HP is obviously inhibited. In another animal experiment, the culture supernatant of probiotics was used, and a similar effect was achieved. In addition, in clinical trials, it was found that lactic acid bacteria culture supernatant, which has been proved to inhibit
HP in vitro, can also partially inhibit human HP infection for a long time in vivo. Therefore,
screening specific strains that can inhibit HP in vitro may have similar effects on human HP
infection.
SUMMARY
Because even different strains of the same species may have great differences in physiological
function and metabolic phenotype, the present invention should screen strains that can inhibit
HP in vivo. According to the invention, a method capable of directly evaluating and inhibiting
HP colonization is established in vitro, which has great significance for the practical application
of the strain of the invention.
According to the invention, Lactobacillus acidophilus LA-10A, which has excellent probiotic
characteristics and can inhibit Helicobacterpylori is screened out. The invention provides a
method for screening strains for inhibiting Helicobacterpylori, which can directly qualitatively
and quantitatively evaluate whether the screened strains can inhibit or eliminate the
colonization of Helicobacterpylori on gastric mucosa cells by using a DNA fluorescent dye
SYTO9 staining method to count the absolute value of adhered cells.
According to one aspect of the present invention, there is provided a method for screening
Helicobacter pylori-inhibiting strains, which further comprises: a test to check the inhibition
effect of bacterial precipitation, culture solution and culture solution supernatant on the growth
of Helicobacterpylori on solid medium, a test to check the ability of preventing healthy mice from being infected with HP bacteria, and a test to check the therapeutic ability of mice infected with HP bacteria.
According to one aspect of the present invention, there is provided a method for screening
Helicobacter pylori-inhibiting strains, and the Lactobacillus acidophilus LA-IOA is screened
by the above method for screening Helicobacterpylori-inhibitingstrains.
According to one aspect of the present invention, there is provided a method for testing the
ability of a strain to inhibit Helicobacter pylori strains, and any of the above methods for
screening and inhibiting Helicobacterpylori strains is used for testing the ability of a strain to
inhibit Helicobacterpylori strains. In vitro, it can inhibit the adhesion (colonization) ability of
Helicobacterpylori in gastric epithelial cells. Animal experiments show that it can reduce or
prevent Helicobacterpylori infection.
DESCRIPTION OF THE FIGURES
Fig. 1 is the morphology of Lactobacillus acidophilus strain in one embodiment of the present
invention.
Fig. 2 shows the inhibitory ability of Lactobacillus acidophilus LA-10A on the growth of HP
on a solid medium in one embodiment of the present invention.
Fig. 3 is a flow cytometric staining standard diagram of Lactobacillus acidophilus LA-10A in
one embodiment of the present invention.
Fig. 4 is a comparison of adhesion ability of Lactobacillus acidophilus LA-10A and
Helicobacterpylorito gastric mucosa cells in one embodiment of the present invention.
Fig. 5 shows the ability to competitively inhibit HP from adhering to gastric mucosal cells when
free LA-OA coexists with HP in one embodiment of the present invention.
Fig. 6 shows the ability of Lactobacillus acidophilus LA-10A to replace HP strains that have
adhered to gastric mucosal cells in one embodiment of the present invention.
Fig. 7 shows the ability of Lactobacillus acidophilus LA-10A to prevent mice from being
infected with HP in one embodiment of the present invention.
DESCRIPTION OF THE INVENTION
The following embodiments are intended to further illustrate some preferred embodiments of
the present invention, but not all of them. Other embodiments based on the present invention
made by professionals in the field without creative labor belong to the scope of the right
protection of the present invention. The present invention will be further explained with
reference to the attached figures.
Embodiment 1: inhibition of Lactobacillus acidophilus LA-10A on the growth of Helicobacter
pylori on solid medium
Inoculate Lactobacillus acidophilus LA-IOA strain into MRS liquid culture medium, culture for
-30 hours, shake well, and take 2mL fermentation broth for later use. The rest fermentation
broth is centrifuged at 10000rpm for 4 minutes, and the bacterial precipitation and supernatant
are collected. The bacterial precipitation is washed twice with PBS buffer, and then the
bacterial precipitation is resuspended with the same volume of sterile PBS buffer, and the
supernatant is collected directly for later use. The HP strain is inoculated in skirrows selective
plate for subculture twice, and inoculated in sterile skirrows liquid medium, cultured for 72
hours, centrifuged at 10000rpm at 4°C for 4 minutes, then collect the bacterial precipitation,
wash the bacterial precipitation twice with PBS buffer, and then resuspend the bacterial
precipitation with PBS buffer, and adjust the absorbance value of the resuspending OD600 to
±0.05 (the corresponding cell concentration is about 101 cells/mL). Take 2mL of HP bacterial precipitation resuspending and put it on the plate, then pour into skirrows solid medium, shake and mix evenly. After the solid culture medium is solidified, 6 pores with a diameter of 0.5cm are made in the solid culture medium, and each pore is sealed with agar. The fermentation broth, bacterial precipitation resuspending and supernatant are injected into the pores of the plate, and the liquid level is flush with the pouring surface of skirrows solid culture medium. Then the plate is cultured in micro-oxygen environment for 72 hours, and the bacteriostatic circle around the pores of the plate is measured. According to the size of bacteriostatic circle, the inhibitory effects of LA-IOA bacterial precipitation, fermentation broth and supernatant on Helicobacterpyloriare tested.
The results show that Lactobacillus acidophilus LA-iOA's fermentation broth, sterile
supernatant and bacterial precipitation resuspending can effectively inhibit HP growth, but the
inhibition ability of sterile supernatant and bacterial precipitation resuspending is weaker than
that of LA-10A fermentation broth. The inhibition ability of LA-1OA's bacterial precipitation
resuspending is about 61% lower than that of LA-IOA fermentation broth and about 46% lower
than that of LA-OA's sterile supernatant, indicating that the bacteriostatic ability of
Lactobacillus acidophilus LA-IOA is the combination of the bacteria itself and its metabolites,
and the maximum inhibitory effect of LA-IOA on HP can be obtained when they work together
(Fig. 2).
Embodiment 2: inhibition test of Lactobacillus acidophilus LA-1OA on adhesion ability of
Helicobacterpylori(HP) to gastric epithelial cells:
In the invention, gastric adenocarcinoma cell SGC7901 is used to simulate gastric mucosal
epithelial cells, and the test of inhibiting the adhesion ability of Helicobacterpylori in gastric
epithelial cells by Lactobacillus acidophilus LA-bOA consists of three tests: (1). Test of the adhesion ability of Lactobacillus acidophilus LA-10A itself to gastric mucosal epithelial cells.
(2). The ability of Lactobacillus acidophilus LA-OA to competitively inhibit HP from adhering
to gastric mucosal cells, when Lactobacillus acidophilus LA-10A coexists with HP. (3). The
substitution effect of Lactobacillusacidophilus LA-10A on HP, when HP cells adhere to gastric
mucosa first. In order to evaluate the adhesion effect, the invention creatively utilizes the
characteristics of the flow cytometer and counts the absolute number of adhered cells by using
the DNA fluorescence dye SYTO9 staining method. SYTO9 dye can bind to DNA or RNA of
bacteria through the cell membrane of dead/living cells. Once it binds to DNA/RNA, its
fluorescence intensity will be greatly enhanced, and it can still retain more than 80% after 3
hours. According to the invention, fluorescence staining is creatively combined with flow
cytometer, so that the absolute number of bacteria adhered to gastric mucosa cells can be
evaluated, and the evaluation result is more credible. It can be directly evaluated qualitatively
and quantitatively whether the selected strains can inhibit or eliminate the colonization of
Helicobacterpylorion gastric mucosal cells.
The test method of the adhesion ability of Lactobacillus acidophilus LA-10A to gastric mucosa:
take 2mL stained LA-10A bacterial suspension, centrifuge, remove supernatant, add 1.5mL
PBS resuspended bacterial precipitation, add the resuspended bacterial precipitation into 12
well plate SGC7901 monolayer cells, culture in an incubator with 5% CO2 at 37C for 1 hour,
and then pour out the nonadherent LA-OA bacterial suspension. Then add 1mL PBS to gently
rinse the SGC7901 cell layer once, then trypsin, EDTA reagent and PBS buffer are added to
digest the adhered SGC7901 cells, and 2mL bacterial suspension is formed again, and then
0.1mL is taken to count the number of cells stained with SYTO9 dye by flow cytometry. Here's
"cell number x20" is the number of Lactobacillus acidophilus LA-10A adhered to SGC7901 cells, and the adhesion rate of Lactobacillus acidophilus LA-10A to gastric cancer cell
SGC7901 can be obtained by taking the measured number of cells in the suspension of LA-IOA
original bacteria as a control.
The test method of the ability of Lactobacillus acidophilus LA-IOA to competitively inhibit HP
from adhering to gastric mucosal cells: take 1mL of stained HP bacterial suspension, centrifuge,
remove the supernatant, add 1mLPBS buffer to resuspend the bacterial precipitation, then mix
with 1mL LA-10A bacterial suspension, add the bacterial suspension to 12-well plate SGC7901
monolayer cells, and culture in an incubator with 5% CO2 at 37C for 1 hour. The unattached
bacterial suspension is poured out, and the SGC7901 cell layer is rinsed once with 1mL PBS,
then trypsin, EDTA reagent and PBS buffer are added to digest the adherent SGC7901 cells,
and 2mL bacterial suspension is reconstituted, and then 0.1mL is taken to count the number of
cells stained with SYTO9 dye by flow cytometry. Here's "cell number x20" is the number of
HP adhering to SGC7901 cells, and the adhesion rate of HP to SGC7901 cells in the presence
of Lactobacillus acidophilus LA-10A can be obtained by taking the measured number of HP
original bacteria suspension cells as a control.
The test method for the ability of Lactobacillus acidophilus LA-iOA to replace HP cells
adhered to gastric mucosa: take 2mL of stained HP bacterial suspension, add it to 12-well plate
SGC7901 monolayer cells, culture it in an incubator with 5%CO2 at 37C for 1 hour, pour out
the non-adhered bacterial suspension, add 1mL PBS to rinse SGC7901 cells gently, and then
add 2mL of LA-1OA bacterial suspension. After being cultured in an incubator with 5%CO2at
37C for 1 hour, the nonadherent bacterial suspension is poured out, and 1mLPBS is added to
rinse SGC7901 cells once, then trypsin, EDTA reagent and PBS buffer are added to digest the
adhered SGC7901 cells, and 2mL bacterial suspension is formed again, and then 0.1mL bacterial suspension is taken to count the number of cells stained with SYTO9 dye by flow cytometry. Here's "cell number x20" is the number of HP adhering to SGC7901 cells. Taking the measured number of HP original bacteria suspension cells as a control, we can evaluate how many adhered HP cells can be replaced by Lactobacillus acidophilus LA-IOA when HP adheres to gastric mucosa cells first.
Test results show that:
Compared with Lactobacillusacidophilus LA-10A, Helicobacterpylori is more likely to adhere
to gastric mucosal cells when only the adhesion ability of the strain itself to gastric mucosal
cells is considered, and the adhesion rate is 21% VS 17.3% (Fig. 4). When the free
Lactobacillus acidophilus LA-10A coexists with HP strain, LA-10A can effectively and
competitively inhibit the adhesion of HP to gastric mucosa cells, which makes the adhesion rate
of HP to gastric mucosa cells decrease by about 53.3% compared with the control (see Fig. 5).
When HP adhered to gastric mucosa cells first, free LA-iOA can also reduce the adhesion rate
of HP, which is decreased by about 33.3% compared with the control (Fig. 6). In summary,
Lactobacillus acidophilus LA-iOA can effectively inhibit or reduce the adhesion of HP to
gastric mucosal cells. Long-term intake of Lactobacillus acidophilus LA-10A can prevent or
reduce the adhesion and colonization ability of HP to gastric mucosa.
8. Therapeutic ability of Lactobacillus acidophilus LA-OA on mice infected with HP
4-6-week-old mice without specific pathogenic bacteria are fed with HP every day for 7
consecutive days. After checking the peripheral blood to confirm that the mice are infected with
HP, all the experimental mice are divided into two groups: control group and experimental
group. In the experimental group, mice are fed drinking water containing Lactobacillus
acidophilus LA-OA 10 9CFU/mL every day for 6 months. The control group is fed with normal diet for 6 months. Then, all mice are taken from peripheral blood for anti-HP-IgG level detection, and then all mice are killed to take gastric antrum for rapid urease detection. The HP cure rate and urease activity of mice are taken as evaluation indexes. The results are shown in
Table 1.
Table 1: Evaluation of therapeutic ability of Lactobacillus acidophilus LA-IOA on mice
infected with HP
HP infection rate Urease activity
Control group 96.2% +++
Experimental group 75%
+ Note: "+" means low urease activity, and "+++" means high urease activity
It can be seen from the experimental results that about 25% of the mice in the experimental
group can not detect HP infection after the intervention of Lactobacillus acidophilus LA-10A
for 6 months, and there is a significant difference in HP infection rate compared with the
control group. Moreover, compared with the control group, the urease activity of gastric antrum
in the experimental group is obviously decreased. It indicated that Lactobacillus acidophilus
LA-10A has a certain healing ability to mice infected with HP. The Lactobacillus acidophilus
strains screened by the screening/detection method are higher than Lactobacillus acidophilus
LA-IOA, and can also be used in the treatment of Helicobacterpylori.
The above embodiments are intended to further illustrate some preferred embodiments of the
present invention, but not all embodiments. Other embodiments based on the present invention
made by professionals in the field without creative labor belong to the scope of the right
protection of the present invention.

Claims (4)

THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS
1. A method for screening Helicobacterpylori-inhibitingstrains, characterized in that the
method for screening Helicobacterpylori-inhibitingstrains includes a method for directly
qualitatively and quantitatively evaluating whether the screened strains can inhibit or
eliminate the colonization of Helicobacter pylori on gastric mucosa cells by using
fluorescent markers and flow cytometry.
2. The method for screening Helicobacterpylori-inhibiting strains according to claim 1,
characterized in that the fluorescent markers and flow cytometry are used to give a
method for directly qualitatively and quantitatively evaluating whether the screened
strains can inhibit or eliminate the colonization of Helicobacterpylorion gastric mucosal
cells, and the DNA fluorescence dye SYTO9 staining method is used to count the
absolute number of adhered cells.
3. The method for screening Helicobacterpylori-inhibiting strains according to claim 2,
characterized in that the method for screening Helicobacter pylori-inhibiting strains
screens out Lactobacillus acidophilus LA-10A, which is preserved in China Center for
Type Culture Collection with the preservation number of CCTCC NO:M2019012.
4. According to claim 3, the Lactobacillus acidophilus LA-10A capable of inhibiting
Helicobacterpylori is used for preparing foods, health products, compound powders and
medicines for inhibiting Helicobacterpylori.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114525231A (en) * 2022-04-20 2022-05-24 微康益生菌(苏州)股份有限公司 Lactobacillus acidophilus for resisting helicobacter pylori infection and culture method and application thereof

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114525231A (en) * 2022-04-20 2022-05-24 微康益生菌(苏州)股份有限公司 Lactobacillus acidophilus for resisting helicobacter pylori infection and culture method and application thereof
CN114525231B (en) * 2022-04-20 2022-07-22 微康益生菌(苏州)股份有限公司 Lactobacillus acidophilus for resisting helicobacter pylori infection and culture method and application thereof

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