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Lactobacillus crispatus for preventing and/or treating genital tract flora disorder related diseases

A technology of Lactobacillus crispatus and bacterial suspension, which is applied in the biological field, can solve the problems of drug resistance of pathogenic bacteria, unfavorable reproductive tract maintenance, and reduced abundance of probiotics, etc., to achieve growth ability acid and alkali resistance, reduce recurrence rate, The effect of improving the cure rate

Pending Publication Date: 2021-03-09
SHENZHEN HUADA GENE INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many problems in antibiotic drug treatment: first, antibiotics kill pathogenic bacteria and eliminate inflammation while inhibiting the growth of probiotics, reducing the abundance of probiotics, which is very unfavorable for the reproductive tract to maintain a stable state, and greatly increases the risk of repeated infection. The risk of infection and recurrence; second, taking drugs cannot achieve the purpose of precise treatment, and can only act on a broad-spectrum systemic range, which is especially easy to damage liver and kidney functions; third, frequent use of antibiotics can easily lead to drug resistance of pathogenic bacteria, This is also an important reason for the secondary infection

Method used

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  • Lactobacillus crispatus for preventing and/or treating genital tract flora disorder related diseases
  • Lactobacillus crispatus for preventing and/or treating genital tract flora disorder related diseases
  • Lactobacillus crispatus for preventing and/or treating genital tract flora disorder related diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1: Isolation and identification of Lactobacillus crispatus LG55-27

[0051] 1. Sample collection

[0052] The isolated sample comes from the feces of a healthy old man. The sample is collected into a sterile sample tube and brought back to the laboratory for sorting within 1 hour.

[0053] 2. Separation and purification

[0054] The collected fresh samples were immediately transferred to an anaerobic operation box, and 0.2g samples were taken in 1mL sterile PBS (phosphate buffer solution), shaken and mixed thoroughly, and then serially diluted and coated. The medium used was a modified version of PYG medium , the specific formula is (1L): tryptone 8g, soybean peptone 2g, polyprotein 1g, casein 1g, yeast powder 10g, beef extract 5g, glucose 5g, K 2 HPO 4 2g, maltose 0.5g, cellobiose 0.5g, soluble starch 0.5g, sodium sulfide 0.25g, Tween 800.5mL, Cysteine-HCl·H 2 O 0.5g, glycerin 0.5mL, sodium acetate 5g, heme 5mg, vitamin K 11μL, inorganic salt solution (eac...

Embodiment 2

[0068] Example 2: Genome sequencing and species classification and functional gene analysis of Lactobacillus crispatus LG55-27

[0069] 1. Genome sequencing

[0070] Centrifuge the overnight cultured LG55-27 bacterial solution at 7,227 g at 4°C for 10 minutes, resuspend the obtained pellet in 1 mL of Tris-EDTA, add 50 μL of 10% SDS and 10 μL of proteinase K (20 mg / mL) at 55°C The cells were lysed in a water bath for 2 hours, and the DNA was extracted by the phenol-chloroform method. And use the Illumina Hiseq 2000 platform to sequence it, the sequencing length is 500bp in both directions, and use SOAPdenovo to assemble the reads. After evaluation, use GCskew to analyze the GC content of the whole genome and visualize the whole genome sequence and functional distribution ( figure 1 ).

[0071] 2. Classification of strain genome species

[0072] Using Checkm software to compare and analyze the whole genome sequence of LG55-27, the most similar species information to this gen...

Embodiment 3

[0075] Embodiment 3: the bioactive substance that Lactobacillus crispatus (Lactobacillus crispatus) LG55-27 produces

[0076] The bioactive substances of LG55-27 mainly examine the content of L-lactic acid and D-lactic acid in the metabolites, and the production of hydrogen peroxide.

[0077] 1. Sample pretreatment

[0078] Strain LG55-27 was inoculated into MRS medium, and cultured at 37°C for 24h under aerobic and anaerobic conditions, respectively.

[0079] Take 1mL of the bacterial solution and centrifuge at 8000r / min for 5min, and take the supernatant to detect the content of L-lactic acid and D-lactic acid.

[0080] Then take 1 mL of the bacterial solution and add lysozyme (final concentration: 1 mg / mL), let it stand at 37°C for 15 min, centrifuge at 8000 r / min for 5 min, take the supernatant, and test the concentration of hydrogen peroxide.

[0081] 2. Measurement method

[0082] The content of L-lactic acid and D-lactic acid is measured by the method in patent CN104...

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Abstract

The invention relates to the technical field of biology, in particular to a novel lactobacillus crispatus strain for preventing and / or treating genital tract flora disorder related diseases and application of the novel lactobacillus crispatus strain. In order to solve the technical problems of different degrees of side effects, high recurrence rate or large trauma and the like of the existing medicine for treating the genital tract flora disorder related diseases, the invention screens out a novel lactobacillus crispatus LG55-27 for preventing and / or treating the genital tract flora disorder related diseases, which can generate secretions such as lactic acid, hydrogen peroxide and the like, is used for inhibiting the growth of pathogenic bacteria of genital tract infection, is used for preventing and / or treating diseases related to genital tract flora disorder, particularly reduces the recurrence rate, has small toxic and side effects and lasting efficacy, and can be used as a dominantbacterial strain of the genital tract to increase the abundance, repair flora disorder and recover vaginal homeostasis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a new strain of Lactobacillus crispatus for preventing and / or treating diseases related to genital flora disturbance and its application. Background technique [0002] The environment of the female reproductive tract is often affected by fluctuations in hormone levels, poor sanitation control, seasonal changes, and even emotional instability, resulting in disordered flora, which in turn leads to the invasion of pathogenic bacteria, causing microbial infections such as bacteria, molds, and viruses. Common There are Escherichia coli, Gardnerella, Candida albicans and HPV viruses. The clinical symptoms of reproductive tract infection are: vaginal itching and burning, abnormal discharge, frequent urination and painful urination, which can lead to premature delivery and cervical cancer in severe cases. According to statistics, the infection rate of reproductive tract microorganisms in wo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/747A61P15/02A61P31/04A61P31/10A61P33/02A61P31/20A61P31/22A23L33/135A23C9/123C12R1/225
CPCA61K35/747A61P15/02A61P31/04A61P31/10A61P33/02A61P31/20A61P31/22A23L33/135A23C9/1234A23V2002/00A23V2200/30
Inventor 张笑薇吕金丽赵少伟高蒙玙邹远强肖亮
Owner SHENZHEN HUADA GENE INST
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