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

A technology of Lactobacillus reuteri and vaginal pathogenic bacteria, applied in the field of microorganisms, can solve the problems of not having the ability to inhibit vaginal pathogenic bacteria, and achieve the effect of superior vaginal epithelial cell adhesion, active and stable biological characteristics

Active Publication Date: 2020-02-14
广东强基药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The present invention aims to provide a specific Lactobacillus reuteri to solve the technical problem that conventional Lactobacillus reuteri does not have the ability to inhibit vaginal pathogenic bacteria in the prior art

Method used

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

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Effect test

Embodiment 1

[0054] Embodiment 1 (isolation, purification, enrichment culture of Lactobacillus reuteri RD-0047 flora)

[0055] 1. Isolation of Lactobacillus reuteri RD-0047 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 in the culture medium containing the prepared MRS medium. dish, 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 reuteri strain RD-0047: 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 a single colony from the inoculation loop A few bacteria in the medium were inoculated into the MRS solid medium according to the "slash method" to obtain isolated and purified single colonies; a sterile to...

Embodiment 2

[0057] Embodiment 2 (identification and preservation of Lactobacillus reuteri RD-0047 bacterial strain)

[0058] 1. Culture characteristics, staining microscopic examination and morphological characteristics: the colonies obtained after culture are as attached figure 1 , the colonies are irregular, round, convex, smooth, and with neat edges; on the blood plate, the colonies are white or milky white; take the smear of the pure culture of the bacteria for Gram staining, and the results are shown in the attached figure 2 , presenting Gram-positive, slightly irregular shape, round-ended Campylobacter, 0.7-1.0 × 2.0-5.0 μm in size, usually present in single, paired, small clusters. The results showed that the isolated strains were preliminarily determined to belong to the genus 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'-TACGGYTACCTTGTT...

Embodiment 3

[0072] Embodiment 3 (determination of metabolites of Lactobacillus reuteri RD-0047)

[0073] 1. Determination of lactic acid content in metabolites of Lactobacillus reuteri RD-0047: 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 The lactic acid of fermented liquid is detected, and the result shows that lactic acid (L / D lactic acid summation) is about 28mg / mL, far higher than the lactic acid content (about 15mg / mL) of Lactobacillus delbrueckii (commercially available), the lactic acid detection collection of the present invention See attached Figure 4

[0074] 2. Determination of hydrogen peroxide content in the metabolites of Lactobacillus reuteri RD-0047: semi-quantitative determination of hydrogen peroxide according to the peroxidase method of Mcgroarty et al., and the isolated and identified Lactobacillus reuteri RD-0047 was inoculate...

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Abstract

The invention provides a Lactobacillus reuteri and its application for preparing vaginal bacteriostatic drugs. The Lactobacillus reuteri is preserved in the General Microbiology Center of China Microbiological Culture Collection Management Committee, and its preservation number is CGMCC No.14110 . The study of its metabolic properties found that, compared with common Lactobacillus reuteri, this strain has stronger lactic acid production ability, hydrogen peroxide production ability, strong antibacterial ability to pathogenic bacteria, and prominent 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 reuteri 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 reuteri, Lactobacillus jansneri, Lactobacillus gasseri, Lactobacillus crispatus, Lactobacillus vaginalis, Lactobacillus rhamnosus, 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 ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N15/31A61K35/747A61P31/04A61P31/02A61P31/10A61P33/02A61P15/02C12R1/225
CPCA61K35/747C12N1/205C12R2001/225Y02A50/30
Inventor 许少燕潘立许锐鹏黄旭藩姚耀宏赵湘江李煜龙
Owner 广东强基药业有限公司
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