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Lactobacillus johnsonii (Ljohn-1) and applications thereof

A Lactobacillus johnsonii technology, applied in the field of microorganisms, can solve the problems of vaginal microecology not returning to a healthy state, microbial resistance, and high recurrence rate, achieving good lactic acid production ability and adhesion to Hela cells, excellent The effect of bacteriostatic ability

Active Publication Date: 2020-02-11
SICHUAN ANAEROBIC BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the antibiotic treatment is effective, it also has great defects. It has the following two aspects: (1) It has an inhibitory effect on all antibiotic-sensitive microorganisms in the vaginal microenvironment. Therefore, the vaginal microecology has not recovered to a healthy, In a state of balance that can resist the invasion of pathogenic bacteria, the suppressed or killed pathogenic microorganisms or foreign pathogenic microorganisms will reproduce again or even cause disease, resulting in recurrence or new vaginal inflammation; (2) Microbes develop drug resistance, and antibiotics Unable to balance the vaginal microecological environment, resulting in refractory BV
Therefore, although antibiotic treatment takes effect quickly, the recurrence rate is high, and the recurrence rate within 3 months is as high as 30%.

Method used

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  • Lactobacillus johnsonii (Ljohn-1) and applications thereof
  • Lactobacillus johnsonii (Ljohn-1) and applications thereof
  • Lactobacillus johnsonii (Ljohn-1) and applications thereof

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Experimental program
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Effect test

Embodiment 1

[0038] The isolation method of Lactobacillus johnsonii described in the present invention comprises the following steps:

[0039] Use vaginal swabs to collect vaginal secretion samples from Chinese women aged 20-40 who have passed the health examination; take 2 mL of sterile anaerobic PBS buffer solution in the anaerobic tube with the above cotton swabs, shake and mix well , and use it as the original solution for ten-fold serial dilution; take 100 μL of the 10000-fold diluted liquid and spread it on the MRS solid medium, and place it in an anaerobic incubator at 37 °C for culture. After 48 hours of culture, pick out a single suspected lactobacillus The colony was cultured in MRS broth medium for 24 hours, and a part of the bacterial liquid obtained after culture was transferred to continue the culture, and the other part of the bacterial liquid was extracted from the bacterial DNA; the bacterial 16S rRNA was amplified and sequenced, and the sequencing results were BLASTed. Fo...

Embodiment 2

[0064] The biochemical identification results of Lactobacillus johnsonii Ljohn-1 are as follows:

[0065] Pass the methyl red test (MR test), acetylmethylmethanol test (VP test), indigo matrix test, aescin hydrolysis test, trisaccharide iron test, Krebs iron test, urease test, phenylalanine Deaminase test, amino acid decarboxylase test, gelatin liquefaction test, sodium malonate test, citrate test (citrate test), nitrate reduction test, litmus milk test, bacterial power test, using French Merieux The API 50 CHL lactobacillus identification system produced by the company carried out biochemical identification of the strain, and the specific results are as follows:

[0066] L. johnsoni Ljohn-1 can hydrolyze aescin to produce glucose and aescin. A negative MR test indicates that the organic acid produced by metabolizing glucose is not enough to change the color of the chromogen. A negative VP test indicates that no pyruvic acid is produced by metabolizing glucose. Indigo matrix ...

Embodiment 3

[0068] Antibiotic susceptibility test of Lactobacillus johnsonii Ljohn-1.

[0069] According to the requirements of the antibiotic susceptibility test in the general introduction of the third part of the Pharmacopoeia of the 2015 edition, the susceptibility of the strains to antibiotics was determined by the agar diffusion disk method, and the susceptibility level of the strains to antibiotics was judged according to the size of the inhibition zone.

[0070] Activation and streaking: Lactobacillus johnsoni Ljohn-1 was activated in MRS broth medium, Escherichia coli was activated in nutrient broth, and cultured at 37 °C; after lactic acid bacteria were activated, a ring of bacterial liquid was picked and placed on MRS solid medium Streak the line, put the plate into an anaerobic sealed tank with an anaerobic gas producing bag, and cultivate it at 37 °C; after the E. Incubate in airtight jars at 37°C.

[0071] Pick several single colonies from the well-cultured agar plate and dir...

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Abstract

The invention discloses Lactobacillus johnsonii (Ljohn-1). Through the utilization of a dominant strain belonging to the vaginas of the women in our country and the utilization of applied technologiesthat the dominant strain can be applied to vaginal micro-ecological preparations, the cure rates of bacterial vaginitis can be increased, and the recurrence rates of the bacterial vaginitis can be reduced. The strain has been deposited with the China Center for Type Culture Collection in June, 2019 and assigned the accession number CCTCC No. M 2019426.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to Lactobacillus johnsonii and its application. Background technique [0002] From the research reports published by the journal "Research Progress and Clinical Significance of Vaginal Microecology" published by the journal Practical Gynecology, and "Research Progress on the Survival Status of Lactobacillus in the Vaginal Microecology" published by the journal China Microecology, we can see that There are more than 300 kinds of microorganisms symbiosis in the human vagina. They restrict each other and check and balance each other to form a dynamic balance. The occurrence of various vaginitis is related to the imbalance of vaginal micro-ecological environment. [0003] The vagina of healthy women of childbearing age is a microenvironment with Lactobacillus as the dominant flora, which can produce H 2 o 2 , lactic acid, lactobacillus, and competitively adhere to the vaginal epithelium,...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/747A61P15/02A61P31/04C12R1/225
CPCA61K35/747A61P15/02A61P31/04C12N1/205C12R2001/225Y02A50/30
Inventor 承磊刘瑶王琼曾婉秋
Owner SICHUAN ANAEROBIC BIOTECHNOLOGY CO LTD
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