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

A technology of Lactobacillus rhamnosus 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 biology Effects of Features

Active Publication Date: 2020-02-14
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 rhamnosus to solve the technical problem that conventional Lactobacillus rhamnosus does not have the ability to inhibit vaginal pathogenic bacteria in the prior art

Method used

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  • A kind of Lactobacillus rhamnosus and its application for preparing vaginal bacteriostatic drugs
  • A kind of Lactobacillus rhamnosus and its application for preparing vaginal bacteriostatic drugs
  • A kind of Lactobacillus rhamnosus 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 rhamnosus RD-0060 flora)

[0055] 1. Isolation and inoculation of Lactobacillus rhamnosus RD-0060 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 A Petri dish of MRS medium, and label the information, place the Petri dish in an anaerobic tank, and put in 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 rhamnosus strain RD-0060: 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 Inoculate a few bacteria in the MRS solid medium according to the "slash method" to obtain isolated and purified single colonie...

Embodiment 2

[0057] Embodiment 2 (identification and preservation of Lactobacillus rhamnosus RD-0060 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 generally milky white, round, with neat edges, smooth surface, semicircular bulges, and opaque; take a smear of the pure culture of the bacteria for Gram staining, and the results are shown in the attached figure 2 , showing Gram-positive, short rod-shaped, blunt-rounded ends, arranged in short chains. The results showed that the isolated strains were 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 determi...

Embodiment 3

[0072] Embodiment 3 (determination of metabolites of Lactobacillus rhamnosus RD-0060)

[0073] 1. Determination of lactic acid content in metabolites of Lactobacillus rhamnosus RD-0060: 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 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 35mg / 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 rhamnosus RD-0060: semi-quantitative determination of hydrogen peroxide according to the peroxidase method of Mcgroarty et al., and inoculate the isolated and identified Lactobacillus rhamnosus RD-0060 at H...

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Abstract

The invention provides a Lactobacillus rhamnosus and its application for preparing vaginal bacteriostatic drugs. The Lactobacillus rhamnosus is preserved in the General Microbiology Center of the China Microbiological Culture Collection Management Committee, and its preservation number is CGMCC No.14184 . The study of its metabolic properties found that, compared with common Lactobacillus rhamnosus, 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 rhamnosus 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 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 bacteria and cause va...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N15/11A61K35/747A61P15/02A61P31/04A61P31/10A61P33/02C12R1/225
CPCA61K2035/115C12N1/205C12R2001/225Y02A50/30
Inventor 许锐鹏许少燕姚耀宏黄旭藩赵湘江李煜龙陈贵浩
Owner 广东龙创基药业有限公司
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