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Staphylococcus epidermidis with broad-spectrum antibacterial activity for gram-positive drug resistant bacteria as well as screening method and application of staphylococcus epidermidis

A Staphylococcus epidermidis, Gram-positive technology, applied in the field of Staphylococcus epidermidis and its screening, can solve the problems of unreported correlation between Staphylococcus epidermidis and anti-Gram-positive drug-resistant bacteria, and is not easy to drug resistance and difficult to produce , targeted effect

Active Publication Date: 2018-11-16
RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • 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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  • Staphylococcus epidermidis with broad-spectrum antibacterial activity for gram-positive drug resistant bacteria as well as screening method and application of staphylococcus epidermidis
  • Staphylococcus epidermidis with broad-spectrum antibacterial activity for gram-positive drug resistant bacteria as well as screening method and application of staphylococcus epidermidis

Examples

Experimental program
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Effect test

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 analysis showed that Staphylococcus accounted for 82.0% of the genus-level culturable 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 and...

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 staphylococcus epidermidis with broad-spectrum antibacterial activity for gram-positive drug resistant bacteria and a screening method of the staphylococcus epidermidis. Thestaphylococcus epidermidis is separated from staphylococcus epidermidis strains colonized in nasal cavity of healthy people, is named N173-2, has obvious bacteriostatic action on clinically common gram-positive drug resistant pathogenic bacteria but has no bacteriostatic action on gram-negative drug resistant bacteria. The genome size of the staphylococcus epidermidis strain N173-2 is 2459364 bp,the staphylococcus epidermidis strain N173-2 contains two plasmids with sizes being 65046 bp and 55156 bp respectively and is preserved in CGMCC (China General Microbiological Culture Collection Center) on April 3, 2018, and the preservation number is CGMCC No. 15550. The invention further relates to an application of the staphylococcus epidermidis N173-2 in preparation of drugs resisting gram-positive drug resistant bacteria. The strain with the broad-spectrum antibacterial activity on the clinically common drug resistant pathogenic bacteria is screened from symbiotic / colonized florae of a human body, and a foundation is laid for exploration of novel antibacterial molecules which are non-toxic and harmless to the human body, brand-new in structure and more targeted and do not easily produce drug resistance.

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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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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