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A kind of Lactobacillus gasseri and its application for preparing vaginal bacteriostatic drugs

A technology of Lactobacillus gasseri and vaginal pathogenic bacteria, applied in the field of microorganisms, can solve the problem of not having the ability to inhibit vaginal pathogenic bacteria, achieve superior vaginal epithelial cell adhesion, inhibit Candida albicans, active and stable biological characteristics Effect

Active Publication Date: 2020-03-10
广东龙创基药业有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention aims to provide a specific Lactobacillus gasseri to solve the technical problem in the prior art that conventional Lactobacillus gasseri does not have the ability to inhibit vaginal pathogenic bacteria

Method used

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  • A kind of Lactobacillus gasseri and its application for preparing vaginal bacteriostatic drugs
  • A kind of Lactobacillus gasseri and its application for preparing vaginal bacteriostatic drugs
  • A kind of Lactobacillus gasseri and its application for preparing vaginal bacteriostatic drugs

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Embodiment 1

[0054] Embodiment 1 (isolation and inoculation, purification, enrichment culture of Lactobacillus gasseri RD-0046 flora)

[0055] 1. Isolation and inoculation of Lactobacillus gasseri RD-0046 flora: Use two sterile cotton swabs to collect the secretions on the side wall of the subject's vagina, and inoculate them with different concentrations within 24 hours in the prepared MRS Petri dish of culture medium, and label the information, place the Petri dish in an anaerobic tank, and put CO 2 Put the gas-producing bag in a 37°C incubator and incubate for more than 48 hours.

[0056] 2. Purification and enrichment culture of Lactobacillus gasseri strain RD-0046: Count the colonies according to their different shapes (surface, edge, etc.) and size, and count the ones with the same shape and size as one, and pick out a single colony from the inoculation loop. A few bacteria were inoculated into the MRS solid medium according to the "slash method" to obtain isolated and purified sing...

Embodiment 2

[0057] Embodiment 2 (identification and preservation of Lactobacillus gasseri RD-0046 bacterial strain)

[0058] 1. Culture characteristics, staining microscopic examination and morphological characteristics: the colonies obtained after culture are as attached figure 1 , the colony is round, with neat edges, smooth surface, relatively flat, and translucent; the smear of the pure culture of the bacteria was taken for Gram staining, and the results are shown in the attached figure 2 , Gram staining was positive, thin rod-shaped, blunt-rounded end, arranged in chains or filaments, the results showed that the isolated strain was preliminarily determined to be Lactobacillus.

[0059] 2. Identification of 16SrDNA gene sequence: DNA was extracted with a bacterial genomic DNA extraction kit, and primers 27F (5'-AGAGTTTGATCMTGGCTCAG-3') and 1492R (5'-TACGGYTACCTTGTTACGACTT-3') were used for PCR amplification. The PCR product was subjected to gel electrophoresis to determine the 16S r...

Embodiment 3

[0072] Embodiment 3 (determination of metabolites of Lactobacillus gasseri RD-0046)

[0073] 1. Determination of lactic acid content in the metabolites of Lactobacillus gasseri RD-0046: adopt high performance liquid phase method (0.005M sulfuric acid aqueous solution (0.28ml sulfuric acid (98%)-1000ml water, pH is about 2.1) to cultivate this bacterial strain for 24-48h to ferment The lactic acid of liquid is detected, and the result indicates that lactic acid (L / D lactic acid sum) is about 35mg / mL, which is much higher than the lactic acid content of commercially available Lactobacillus delbrueckii (about 15mg / mL) and conventional Lactobacillus gasseri. Lactic acid detection map see attached Figure 4

[0074] 2. Determination of hydrogen peroxide content in the metabolites of Lactobacillus gasseri RD-0046: semi-quantitative determination of hydrogen peroxide was carried out according to the peroxidase method of Mcgroarty et al., and the isolated and identified Lactobacillus...

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Abstract

The invention provides a Lactobacillus gasseri and its application for preparing vaginal bacteriostatic drugs. The Lactobacillus gasseri is preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee, and its preservation number is CGMCC No.14109. The study of its metabolic properties found that compared with Lactobacillus vulgaris, this strain has stronger lactic acid and hydrogen peroxide production capacity, strong antibacterial ability against pathogenic bacteria, and outstanding vaginal epithelial cell adhesion ability. Based on the new properties discovered above, the present invention determines the new application of using it to prepare vaginal antibacterial drugs, which can realize the treatment of various bacterial vaginal diseases. The bacterial agent of the invention has remarkable effects on bacterial vaginal diseases, is safe, non-toxic, has good stability and can be stored for a long time, and also relates to its use in medicines for preventing and / or treating gynecological diseases.

Description

technical field [0001] The invention relates to the technical field of microorganisms, further relates to the regulation of vaginal micro-ecological environment and the treatment of bacterial vaginal diseases, in particular to a kind of Lactobacillus gasseri and its application for preparing vaginal bacteriostatic drugs. Background technique [0002] There are a variety of microorganisms in the vagina of healthy women, and they form a vaginal micro-ecological system that restricts, coordinates and dynamically balances with the host and the environment. The vaginal flora of healthy women is mainly composed of Lactobacillus, including Lactobacillus plantarum, Lactobacillus jansneri, Lactobacillus gasseri, Lactobacillus crispatus, Lactobacillus vaginal, Lactobacillus gasseri, etc. Under normal circumstances, Lactobacillus can protect the vagina. When the Lactobacillus in the vaginal microecology is disturbed, it can lead to the invasion of pathogenic bacteria and cause vaginiti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/747A61P31/02A61P31/04A61P31/10A61P15/02A61P33/02C12R1/225
CPCA61K2035/115C12N1/205C12R2001/225Y02A50/30
Inventor 李煜龙陈贵浩潘立黄旭藩许少燕姚耀宏赵湘江
Owner 广东龙创基药业有限公司
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