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Lactobacillus crispatus for preventing and treating female urogenital tract infection and application thereof

A technology of Lactobacillus crispatus and microbial strains, applied in the field of microorganisms, can solve the problems of increasing dosage, inevitable inconvenience in the use process, and not very strong adhesion, and achieves the effects of strong antibacterial ability and strong adhesion.

Active Publication Date: 2020-11-06
哈尔滨美华生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Both of the above two bacteria have a certain adhesion ability of vaginal epithelial cells to ensure that they can colonize the vagina, but their adhesion ability is not very strong, and the application needs to increase the dosage to achieve the therapeutic effect.
[0008] And at present, most Lactobacillus crispatus is directly administered through the vagina to treat vaginal inflammation, which is inevitably inconvenient during use

Method used

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  • Lactobacillus crispatus for preventing and treating female urogenital tract infection and application thereof
  • Lactobacillus crispatus for preventing and treating female urogenital tract infection and application thereof
  • Lactobacillus crispatus for preventing and treating female urogenital tract infection and application thereof

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0072] Basic experiment example 1 Isolation of Lactobacillus crispatus strain

[0073] The composition of MRS medium is as follows: peptone 10.0g; beef extract 10.0g; yeast extract 5.0g; diammonium hydrogen citrate 2.0g; glucose 20.0g; Tween 80, 1.0mL; sodium acetate (CH 3 COONa·3H 2 O) 5.0g; Dipotassium hydrogen phosphate (K 2 HPO 4 ·3H 2 O) 2.0g; Magnesium sulfate (MgSO 4 ·7H 2 O) 0.58g; manganese sulfate (MnSO 4 ·H 2 O) 0.25g; distilled water 1000mL; adjusted to pH6.2-6.6.

[0074] The separation steps are as follows:

[0075] (1) Collect 100 samples of vaginal secretions from healthy women without clinical symptoms and signs of vaginitis, number the samples sequentially, inoculate them in MRS liquid medium, and culture them anaerobically at 37°C for 24 hours.

[0076] (2) For each sample, 5 single colonies were randomly selected and cultured in 2 mL MRS liquid medium for 24 hours, and the colonies were numbered sequentially.

[0077] (3) Amplify bacteria for bact...

experiment example 2

[0078] Basic experiment example 2 Identification of Lactobacillus crispatus strains

[0079] The bacterial strain Lcr-MH175 screened in Basic Experimental Example 1 was identified as follows:

[0080] (1) Morphological identification: On the MRS Lactobacillus medium plate, the isolated strain Lcr-MH175 appeared as opaque, protruding milky white colonies with a size of about 1-2 mm, and the edges of the colonies were neat and moist, and the size and shape were stable. ,See figure 1 A in A; Gram-positive, short rod-shaped, can be connected into a chain, see figure 1 B in

[0081] (2) 16s rDNA sequence homology analysis

[0082] The above-mentioned isolated bacterial strain Lcr-MH175 was cultivated by conventional methods, the total DNA of the bacterial strain was extracted as a template for gene amplification, and the bacterial 16S rDNA gene conservation Area.

[0083] The amplification system (25 μL) is: 1×PCR reaction buffer, 200 μmol / L dNTPs, 0.2 μmol / L upstream and down...

Embodiment 1

[0086] Example 1 Lactobacillus crispatus Lcr-MH175 biochemical reaction experiment

[0087] Physiological and biochemical tests were performed on Lactobacillus crispatus Lcr-MH175, including catalase test, gelatin liquefaction test, H 2 S test, nitrate reduction test and exercise test for identification:

[0088] 1. Catalase test

[0089] Pick out a single purified colony, place it on a new glass slide, and add a few drops of 0.3% hydrogen oxide solution. As a result, on the contrary, no bubbles are generated as a negative (-) result.

[0090] 2. Gelatin liquefaction experiment

[0091] Pick the purified single colony with an inoculation needle and inoculate it on a gelatin solid medium by puncture, the puncture depth is about 2 / 3 of the medium, culture in an incubator at 20°C for 7 days, take it out every 24h, put it in a refrigerator at 4°C for 30min, and then take it out again. It is a positive (+) result if it does not coagulate within a certain period of time, and a neg...

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Abstract

The invention discloses lactobacillus crispatus Lcr-MH175 for preventing and treating female urogenital tract infection. The preservation number of the Lcr-MH175 is CGMCC No. 15938. The lactobacilluscrispatus Lcr-MH175 provided by the invention has relatively strong antibacterial ability, can effectively inhibit growth of external pathogenic bacteria and pathogenic bacteria under other conditions, and also has a relatively strong adhesion effect on Hela cells so as to ensure the colonization of the strain on vaginal epithelium. The lactobacillus crispatus Lcr-MH175 provided by the invention has a certain application prospect in the aspect of urogenital tract infection prevention and treatment.

Description

technical field [0001] The invention belongs to the field of microbes, and in particular relates to a strain of Lactobacillus crispatus for preventing and treating women's urogenital tract infection and its application. Background technique [0002] Urogenital tract infection is a common disease that seriously affects the physical and mental health of women. Most of them are Escherichia coli or Staphylococcus aureus and Candida albicans infection. The prevalence rate is as high as 40%. Its treatment mainly relies on antibiotics, but the cure rate is low. The rate can reach more than 40%. [0003] 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,...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/04C12N11/10C12N11/04A61K35/747A23L33/135A61P31/04A61P31/10C12R1/225
CPCA23V2002/00A61K35/747A23L33/135A61P31/04A61P31/10C12N1/04C12N1/20C12N11/04C12N11/10C12N1/205C12R2001/225A23V2200/30
Inventor 肖雯娟
Owner 哈尔滨美华生物技术股份有限公司
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