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A strain of Staphylococcus epidermidis with broad-spectrum antibacterial activity against Gram-positive drug-resistant bacteria and its screening method and application

A Staphylococcus epidermidis, Gram-positive technology, applied in the field of Staphylococcus epidermidis and its screening, can solve the problems of unreported association between Staphylococcus epidermidis and anti-Gram-positive drug-resistant bacteria, and achieves difficult drug resistance, difficult to produce , targeted effect

Active Publication Date: 2021-10-22
RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no relevant literature reporting the association between Staphylococcus epidermidis and resistance to Gram-positive drug-resistant bacteria

Method used

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  • A strain of Staphylococcus epidermidis with broad-spectrum antibacterial activity against Gram-positive drug-resistant bacteria and its screening method and application
  • A strain of Staphylococcus epidermidis with broad-spectrum antibacterial activity against Gram-positive drug-resistant bacteria and its screening method and application

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Experimental program
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Embodiment 1

[0029] This embodiment is a screening procedure for Staphylococcus epidermidis with broad-spectrum antibacterial activity against Gram-positive drug-resistant bacteria, which includes:

[0030] Step 1) Perform routine flora culture on the nasal swab samples of 200 males and 200 females aged 18-25 years old with normal physical examination, no previous disease history, and no hospital contact history in the past six months in Shanghai, and 20 monoclonal samples for each sample Identification by MALDI-TOF MS and 16S rRNA gene sequencing. The results of the analysis showed that Staphylococcus accounted for 82.0% of the genus-level cultivable flora in the nasal cavity of healthy people; further identification of Staphylococcus bacteria found that Staphylococcus epidermidis accounted for 79.8%.

[0031] Step 2) Analyze the pathogenic methicillin-resistant Staphylococcus aureus (MRSA) and macrolide-resistant staphylococcus aureus (MRSA) isolated from different countries, communities...

Embodiment 2

[0037] In this example, the gene sequencing of Staphylococcus epidermidis N173-2, a molecule with broad-spectrum antibacterial activity of clinical Gram-positive drug-resistant pathogenic bacteria screened in Example 1, was carried out. The result is as follows:

[0038] In this embodiment, the whole genome sequencing analysis of Staphylococcus epidermidis strain N173-2, which produces molecules with broad-spectrum antibacterial activity against clinical Gram-positive drug-resistant pathogens ( figure 2 Part A of the study), it was found that its size was 2459364bp, its G+C content was 32%, its MLST type was ST73, and it contained 4 Prophage (prophage), 3 GI-like regions (genome islands), 2 plasmids, each The sizes of the plasmids were 65046bp and 55156bp ( figure 2 Part B of the above); and the phylogenetic analysis of this bacterium based on the phylogenetic tree of the core SNP of the whole genome was carried out by the kSNP 3.0 software, and it was found that the evolut...

Embodiment 3

[0041] This embodiment is the antibacterial experiment verification of Staphylococcus epidermidis strain N173-2, and it takes Staphylococcus aureus USA300 as an example, and the verification steps are as follows:

[0042] (1) Pick a single clone of Staphylococcus aureus USA300 and inoculate 10 ml of fresh TSB, shake the bacteria overnight at 37°C and 220 rpm.

[0043] (2) Detect the OD600 of the bacteria with a microplate reader, adjust the OD600 of the bacteria to 0.0001 and inoculate them into 50 ml of solid TSB medium, mix well and spread the plates.

[0044] (3) Pick a single clone of the N173-2 strain isolated from the nasal cavity of a healthy person and inoculate it into 10 ml of fresh TSB, shake the bacteria overnight at 37°C and 220 rpm.

[0045] (4) Take 5 μl of the bacterial suspension, the centrifuged supernatant and the precipitate of the bacterial culture and inoculate them on a solid plate containing USA300 bacteria. After drying, place them in a 37°C incubator ...

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Abstract

The invention discloses a strain of Staphylococcus epidermidis with broad-spectrum antibacterial activity against Gram-positive drug-resistant bacteria and a screening method thereof. The Staphylococcus epidermidis is isolated from a healthy human nasal cavity colonized Staphylococcus epidermidis strain, named N173-2, and is clinically effective. Common gram-positive drug-resistant pathogens have obvious antibacterial effect, but no bacteriostatic effect on gram-negative drug-resistant bacteria. The genome size of the Staphylococcus epidermidis strain N173-2 is 2459364bp, including 2 plasmids, the sizes are 65046bp and 55156bp respectively, and it has been deposited in the General Microorganism Center (CGMCC) of the China Committee for Culture Collection of Microorganisms on April 03, 2018. Its deposit number is CGMCC No.15550. The present invention also relates to the application of the Staphylococcus epidermidis N173‑2 in the preparation of anti-Gram-positive drug-resistant bacteria drugs. The present invention screens the strains with broad-spectrum antibacterial activity against common clinical drug-resistant pathogenic bacteria from the symbiotic / colonization flora of human body. Molecules lay the groundwork.

Description

technical field [0001] The invention relates to the field of microbial animal cell lines, in particular to a Staphylococcus epidermidis strain with broad-spectrum antibacterial activity against Gram-positive drug-resistant bacteria and a screening method thereof. Background technique [0002] Multi-drug resistant bacterial infection has become a thorny public health issue worldwide. In 2017, WHO issued a global warning on 12 clinically resistant bacteria, carbapenem-resistant Enterobacteriaceae (CRE), vancomycin-resistant Enterococcus (VRE) and methicillin-resistant Staphylococcus aureus (MRSA) etc. are the most difficult pathogenic types for clinical anti-infection treatment. Most of the existing antibiotics are derived from the metabolites of environmental (soil, etc.) microorganisms, but this limited resource of environmental microorganisms has been exploited to the greatest extent in the 1960s. With the progress of human microbiome research in recent years, researchers...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/741A61P31/04C12R1/45
CPCA61K2035/115A61P31/04C12N1/205C12R2001/45
Inventor 李敏刘倩孟红委刘瑶刘俊兰汪骅
Owner RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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