Lactobacillus crispatus and its application in gynecological diseases

A Lactobacillus crispatus, vaginal disease technology, applied in application, sexual diseases, resistance to vector-borne diseases, etc., can solve the problems of unable to maintain viable bacterial content, poor colonization ability, unable to meet gynecological clinical conditions, and achieve superior vaginal epithelial cells. Adhesion, Inhibition of Candida albicans, Active Stabilization of Biological Properties

Active Publication Date: 2014-12-03
SUZHOU OSEL BIO PHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the existing products on the market are not the dominant vaginal flora of women in my country, the colonization

Method used

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  • Lactobacillus crispatus and its application in gynecological diseases
  • Lactobacillus crispatus and its application in gynecological diseases
  • Lactobacillus crispatus and its application in gynecological diseases

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Experimental program
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Effect test

Embodiment 1

[0055] Embodiment 1, Isolation, inoculation, purification and enrichment culture of Lactobacillus crispatus 262-1

[0056] 1. Isolation and inoculation of Lactobacillus crispatus 262-1 flora: samples were collected using Port. A-Cd system. Use two sterile cotton swabs to collect 1 / 3 of the secretions on the side wall of the subject's vagina, inoculate them in different concentrations on a petri dish containing the prepared Rogosa SL medium within 24 hours, and mark the information. The dish was placed in an anaerobic tank and placed in CO 2 Put the gas-producing bag in a 37°C incubator and incubate for more than 48 hours.

[0057] 2. Purification and enrichment culture of Lactobacillus crispatus strain 262-1: 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 few of a single colony with the inoculation loop Bacteria are inoculated into MRS solid medium according to the "...

Embodiment 2

[0058] Embodiment 2, Identification and preservation of Lactobacillus crispatus 262-1 strain

[0059] 1. Culture characteristics, staining microscopic examination and morphological characteristics: The colonies obtained after cultivation are shown in Figure 1. The colonies are gray-white round, full in the middle, diffuse around, and irregular; take a smear of the pure culture of the bacteria for Gram staining , the result is as figure 2 , showing Gram-positive, short rods, which can be connected to long chains; the results of electron microscope analysis are shown in Figure 3. Under the electron microscope, the strain has no spores, no flagella, no capsule, and the strain size is 26.824×6.667um. The results showed that the isolated strains were preliminarily determined to belong to the genus Lactobacillus.

[0060] 2. Identification of 16SrDNA gene sequence: DNA extraction was carried out with bacterial genomic DNA extraction kit, and primer pair 8F (5'-AGA GTT TGATCC TGG...

Embodiment 3

[0064] Embodiment 3, Determination of Metabolites of Lactobacillus crispatus 262-1

[0065] one. Determination of lactic acid content in metabolites of Lactobacillus crispatus 262-1: The D-lactic acid detection kit measured the output of D-lactic acid of this strain, and the result was 6.213g / L; the content of L-lactic acid measured by the sensor analyzer was 3.789 g / L, the results are shown in the T0 group data in Table 1 below.

[0066] Table 1 Determination results of lactic acid

[0067] sample D-lactic acid (g / L) L-lactic acid (g / L) T0 6.213 3789 T30 6.334 3.330 T50 6.291 3.225

[0068] two. Determination of hydrogen peroxide content in metabolites of Lactobacillus crispatus 262-1: Semi-quantitative determination of hydrogen peroxide was carried out according to the peroxidase method of Mcgroarty et al. The isolated and identified Lactobacillus crispatus 262-1 was inoculated in H 2 o 2 To identify the MRS-TMB plate, after anaer...

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Abstract

The invention discloses a Lactobacillus crispatus 262-1 and its application in gynecological diseases. The Lactobacillus crispatus 262-1 is a new Lactobacillus crispatus strain, and has a preservation number of CGMCCNo.6469 in China General Microbiological Culture Collection Center. The Lactobacillus crispatus 262-1 has the advantages of strong acid and H2O2 production ability, strong adherence to vaginal epithelial cells, substantial resistance on bacterial vaginal diseases and various vaginal gynecological diseases, safety, non-toxicity, good stability and long term preservation. The invention also relates to a use of the Lactobacillus crispatus 262-1 in medicines for preventing and/or treating the gynecological diseases.

Description

technical field [0001] The present invention relates to a new bacterial strain of the genus Lactobacillus and its application, in particular to a kind of Lactobacillus crispatus ( Lactobacillus crispatus ), which can significantly inhibit Candida albicans and pathogenic bacteria while regulating the vaginal microenvironment and inhibiting Gardnerella vaginalis (BV pathogenic bacteria). Potential for unique therapeutic advantages. 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 crispatus, Lactobacillus jansneri and Lactobacillus gasseri. Under normal circumstances, Lactobacillus can protect the vagina, but the disturbance of vaginal microecology in which Lactobacillus dominates can lead to vagin...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N15/31A61K35/74A61P31/00A61P31/04A61P31/10A61P15/00C12R1/225
CPCY02A50/30
Inventor 刘阳
Owner SUZHOU OSEL BIO PHARM
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