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Lactobacillus crispatus and application thereof

A technology of Lactobacillus crispatus and bacteria agent, which is applied in the field of microorganisms, can solve the problems of increased dosage and weak adhesion ability, and achieve the effects of inhibiting reproduction, strong ability to produce lactic acid, and excellent antibacterial ability

Pending Publication Date: 2021-04-27
SICHUAN ANAEROBIC BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] Although the three types of Lactobacillus crispatus mentioned in the above-mentioned patent documents all have a certain adhesion ability of vaginal epithelial cells and can colonize the vagina, but because of their relatively weak adhesion ability, they need to increase the dosage in application. achieve therapeutic effect

Method used

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  • Lactobacillus crispatus and application thereof
  • Lactobacillus crispatus and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0042] The preparation of embodiment 1 bacterial culture medium

[0043] 1. Preparation of MRS Broth:

[0044] Weigh 52.0g of MRS finished medium powder, dissolve it in 1L distilled water; heat to boil, cool to room temperature, add 0.55g cysteine ​​hydrochloride, stir and dissolve, adjust the pH value to 6.5; install a quantitative dispenser and pass N 2 , heated to boiling, boiled for 20 minutes in a slightly boiling state, cooled and dispensed into 10mL anaerobic tubes, sterilized at 118°C for 20 minutes, stored in a cool place and protected from light, and used for later use.

[0045] 2. Preparation of MRS solid medium:

[0046] Weigh 52.0g of MRS finished medium powder and 15.0g of agar powder, dissolve them in 1L of distilled water, heat to boil, add 0.55g of cysteine ​​hydrochloride after boiling, adjust the pH value to 6.5, and extinguish with high temperature and damp heat at 118°C Bacteria for 20 minutes, store in the shade and away from light, and set aside.

[...

Embodiment 2

[0059] Isolation and screening of embodiment 2 Lactobacillus crispatus

[0060] 1. Isolation of Lactobacillus crispatus:

[0061] Vaginal swabs were used to collect vaginal secretion samples from Chinese women aged 20 to 40 who had passed the physical examination; 2 mL of sterile anaerobic PBS buffer solution was placed in the anaerobic tube containing the above cotton swabs, fully Shake and mix well, and use it as the original solution for continuous 10-fold gradient dilution; take 100 μL of the 10,000-fold diluted liquid and spread it on the MRS solid medium, place it in an anaerobic incubator, and cultivate it at 37°C. After culturing for 48 hours, pick A single colony of suspected Lactobacillus was cultured in MRS broth for 24 hours, and a part of the bacterial liquid obtained after culture was transferred to continue the culture, and the other part of the bacterial liquid was extracted from the bacterial DNA; the bacterial 16S rRNA was amplified and sequenced, and the seq...

Embodiment 3

[0093] Example 3 strain identification

[0094] Lactobacillus crispatus Lcris-18 was cultured and collected, genomic DNA was extracted, and the 16SrRNA sequence was amplified by primers 27F and 1492R to obtain a PCR product, which was then sequenced.

[0095] Sequencing results showed that the sequence of the PCR product was shown as SEQ ID No.1, which was identified as coiled-coil after comparison and analysis on the NCBI website (https: / / blast.ncbi.nlm.nih.gov / Blast.cgi) Lactobacillus.

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Abstract

The invention relates to the technical field of microorganisms, and discloses lactobacillus crispatus and application thereof. The lactobacillus crispatus has the advantage of cell adhesion, so that the lactobacillus crispatus can be adhered to vaginal epithelial cells, a micro-ecological barrier is formed, and pathogenic bacteria are prevented from colonizing or competing for epithelial cell receptors; meanwhile, the lactic acid producing capacity is high, the pH can be reduced, and moreover, generated antibacterial substances, such as hydrogen peroxide, lactic acid, bacteriocin and the like, can also inhibit the growth of the various pathogenic bacteria, such as gardnerella vaginalis, staphylococcus aureus, pseudomonas aeruginosa, escherichia coli, salmonella paratyphi B, shigella dysenteriae and the like.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a lactobacillus crispatus and its application. Background technique [0002] At present, female vaginal infection has become one of the most serious infectious diseases in the world, and millions of women around the world suffer from this disease every year. The female vagina is a complex micro-ecological system, which is composed of the anatomical structure of the vagina, micro-ecological flora, local immunity and the body's endocrine regulation function. Diseases are dominated by three factors: etiology, environment and host, but the core is normal Interrelationships between microbiota and their hosts. From a clinical point of view, the female reproductive tract microecology is closely related to reproductive tract infection. The incidence of various vaginitis is mostly related to the imbalance of vaginal microecology. Bacterial vaginosis (Bacterial vaginosis, BV) is d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/747A61P15/02A61P31/04C12R1/225
CPCC12N1/20A61K35/747A61P15/02A61P31/04Y02A50/30
Inventor 刘瑶王琼曾婉秋高翔
Owner SICHUAN ANAEROBIC BIOTECHNOLOGY CO LTD
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