A kind of Lactobacillus crispatus and its application in gynecological diseases

A technology for Lactobacillus crispatus and vaginal diseases, applied in the fields of application, diseases, and diseases transmitted by vectors, etc., can solve the problems of inability to maintain the content of viable bacteria, poor colonization ability, and inability to meet gynecological clinical conditions, and achieve superior vaginal epithelial cells Adhesion, inhibition of Candida albicans, active and stable biological properties

Active Publication Date: 2016-08-24
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 ability is poor, and the content of viable bacteria cannot be maintained stably, which cannot meet the clinical needs of gynecology.

Method used

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  • A kind of Lactobacillus crispatus and its application in gynecological diseases
  • A kind of Lactobacillus crispatus and its application in gynecological diseases
  • A kind of Lactobacillus crispatus and its application in gynecological diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1. Isolation, inoculation, purification, enrichment culture of Lactobacillus crispatus 262-1 flora

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

[0057] 2. Purification and enrichment culture of Lactobacillus crispatus strain 262-1: count according to the different shapes (surface, edge, etc.) and size of the colonies. The same shape and size are counted as one type, and a few of the single colonies are picked from the inoculation loop Bacteria are inoculated into MRS solid medium...

Embodiment 2

[0058] Example 2. Identification and preservation of Lactobacillus crispatus 262-1 strain

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

[0060] 2. 16SrDNA gene sequence identification: DNA extraction with bacterial genomic DNA extraction kit, and primer set 8F (5'-AGA GTT TGATCC TGG CTC AG-3'), 926R (5'-CCG TCAATT CCTTTR AGTTT-3') ), where R re...

Embodiment 3

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

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

[0066] Table 1 Results of lactic acid measurement

[0067] sample

D-lactic acid (g / L)

L-lactic acid (g / L)

T0

6.213

3.789

T30

6.334

3.330

T50

6.291

3.225

[0068] two. Determination of hydrogen peroxide content in the metabolites of Lactobacillus crispatus 262-1: According to the peroxidase method of Mcgroarty et al. for semi-quantitative determination of hydrogen peroxide, the isolated and identified Lactobacillus crispatus 262-1 was inoculated into H 2 O 2 The MRS-TMB plate was identified, and after anaerobic culture at 37°C for 24 hours, the plate was taken...

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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 invention relates to a new strain of the genus Lactobacillus and its application, in particular to a Lactobacillus crispatus (Lactobacillus crispatus), which regulates the vaginal microenvironment and inhibits Gardnerella vaginalis (BV pathogenic bacteria) while having Obviously inhibits Candida albicans and pathogenic bacteria, and has the potential of unique therapeutic advantages for the most common clinical bacterial vaginitis with fungal infection. Background technique [0002] There are a variety of microorganisms in the vagina of healthy women, and they form a vaginal microecological system that restricts each other, coordinates with each other, and dynamically balances with the host and the environment. The vaginal flora of healthy women is mainly composed of Lactobacillus, including Lactobacillus crispatus, Lactobacillus jannaschii and Lactobacillus gasseri. Under normal circumstances, lactobacilli can play a protective role in the vagina, and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N15/31A61K35/747A61P31/00A61P31/04A61P31/10A61P15/00C12R1/225
CPCY02A50/30
Inventor 刘阳
Owner SUZHOU OSEL BIO PHARM
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