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

A Staphylococcus, Gram-positive technology, applied in the field of human Staphylococcus and its screening, can solve the unreported problems of the correlation between Staphylococcus and anti-Gram-positive drug-resistant bacteria, and achieve drug resistance, new structure, highly targeted effects

Active Publication Date: 2021-10-22
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 human Staphylococcus and anti-Gram-positive drug-resistant bacteria

Method used

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

Examples

Experimental program
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Embodiment 1

[0029] This embodiment is a screening step for human staphylococcus with broad-spectrum antibacterial activity to Gram-positive drug-resistant bacteria, which includes:

[0030] Step 1) Perform routine flora culture on the skin swab samples of 200 males and 200 females aged 18-25 years old with normal physical examinations, 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 found that Staphylococcus accounted for 80.2% of the skin cultureable flora of healthy people at the genus level. Further identification of Staphylococcus species was carried out, and it was found that Staphylococcus hominis accounted for 46.4%.

[0031] Step 2) Analyze the pathogenic methicillin-resistant Staphylococcus aureus (MRSA) and macrocyclic endogenous staphylococcus isolated from the skin surface of healthy people by plate...

Embodiment 2

[0037] In this example, the gene sequencing of Staphylococcus hominis S34-1, a molecule with broad-spectrum antibacterial activity among clinical Gram-positive drug-resistant pathogens screened in Example 1, was carried out. The result is as follows:

[0038] In this embodiment, the human staphylococcus strain S34-1, which produces molecules with broad-spectrum antibacterial activity against clinical Gram-positive drug-resistant pathogens, is subjected to whole-genome sequencing analysis ( figure 2 A), found that its size is 2195430bp, G+C content is 50%, contains 7 Prophage (prophage), 3 GI-like region (genome island), 2 plasmids, the size is 29577bp and 29535bp respectively ( figure 2 B).

[0039] The human staphylococcus strain S34-1 described in this example has been preserved on April 3, 2018, and its classification is named Staphylococcus hominis (Staphylococcus hominis), and its preservation number is CGMCC No.15552, and the preservation unit is China Microbiology G...

Embodiment 3

[0041] This embodiment is the antibacterial experiment verification of human staphylococcus bacterial strain S34-1, 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) Detection of bacterial OD with a microplate reader 600 , to adjust the bacterial OD 600 Inoculate to 0.0001 into 50ml solid TSB medium, mix well and spread the plate.

[0044] (3) Pick a single clone of the S34-1 strain isolated from healthy human skin 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 for overnight cultivation, and...

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Abstract

The invention discloses a strain of human staphylococcus with broad-spectrum antibacterial activity against Gram-positive drug-resistant bacteria and a screening method thereof. 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 human staphylococcus strain S34-1 is 2195430bp, including 2 plasmids, the size is 29577bp and 29535bp respectively, and it has been deposited in the General Microbiology Center (CGMCC) of the China Committee for Microbial Culture Collection on April 03, 2018. Its deposit number is CGMCC No.15552. The present invention also relates to the application of the human staphylococcus S34‑1 in the preparation of anti-Gram-positive drug-resistant bacteria. 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 human staphylococcus 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 have ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12Q1/6869C12Q1/14A61K35/741A61P31/04C12R1/44
CPCA61K35/74A61P31/04C12Q1/6869C12N1/205C12R2001/44
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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