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Staphylococcus hominis with broad-spectrum antibacterial activity on gram-positive drug-resistant bacteria and screening method and application thereof

A Staphylococcus and Gram-positive technology, which is 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 and strong pertinence. , new structural effects

Active Publication Date: 2018-11-06
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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  • Staphylococcus hominis with broad-spectrum antibacterial activity on gram-positive drug-resistant bacteria and screening method and application thereof
  • Staphylococcus hominis with broad-spectrum antibacterial activity on gram-positive drug-resistant bacteria and screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

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 example, 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 03, 2018, and its classification is named Staphylococcus hominis (Staphylococcus hominis), and its preservation number is CGMCC No.15552, and the depository unit is China Microbiology Gener...

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 staphylococcus hominis with broad-spectrum antibacterial activity on gram-positive drug-resistant bacteria and a screening method thereof. The staphylococcus hominis is separated from the skin of a healthy person and is colonized; the name of the staphylococcus hominis is S34-1; the staphylococcus hominis has obvious bacteriostatic effect on clinically common gram-positivedrug-resistant pathogens, but has no bacteriostatic effect on gram-negative drug-resistant bacteria. A genome of the staphylococcus hominis S34-1 is 2195430bp, and the staphylococcus hominis S34-1 contains two plasmids, namely 29577bp and 29535bp; the staphylococcus hominis S34-1 has been collected into CGMCC (China General Microbiological Culture Collection Center) on April 3rd, 2018; the collection number is CGMCC No.15552. The invention also relates to the application of the staphylococcus hominis S34-1 to preparation of gram-positive drug-resistant bacteria drugs. The staphylococcus hominis has the advantages that the bacteria strain with broad antibacterial activity on the clinically common drug-resistant pathogens is separated from the symbiosis / colonization flora of the human body,and a foundation is laid for researching of novel antimicrobial molecules with low toxicity and no harm to human body, fully-new structure, less resistance to drug and stronger pertinence.

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 Applications(China)
IPC IPC(8): C12N1/20C12Q1/6869C12Q1/04A61K35/74A61P31/04C12R1/44
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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