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Phage and application thereof

A bacteriophage and microbial strain technology, applied in the field of biomedicine, can solve problems such as drug resistance of Serratia marcescens, and achieve the effect of overcoming drug resistance problems and treating infections

Active Publication Date: 2021-01-08
SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of this application is to provide a kind of phage and its application, aiming to solve the technical problem of the drug resistance of Serratia marcescens

Method used

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  • Phage and application thereof
  • Phage and application thereof
  • Phage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Screening and purification of embodiment 1 phage

[0030] 1. Sewage treatment

[0031] Take the sewage from a certain river in Shenzhen, centrifuge it at 4000rpm for 15min, filter it with a 0.22μm filter membrane, and put the filtrate into a sterile culture bottle.

[0032] 2. Phage culture

[0033] Take 4 mL of the filtered supernatant, add 200 μL of logarithmic phase host bacterial culture solution, place in an incubator at 37°C and 220 rpm for 3 hours, and filter with a microporous membrane to obtain the phage stock solution.

[0034] 3. Single phage screening and identification

[0035] Pick phage plaques with the same shape and size on the double-layer agar plate, puncture the selected phage plaques with a sterile inoculation needle, insert 3mL LB (Luria-Bertani) broth medium, Stir gently for several times, add 0.2mL of Serratia marcescens bacteria solution, shake well, let stand at room temperature for 1h, incubate at 37°C for 12h, centrifuge at 12000×g for 30m...

Embodiment 2

[0042] Example 2 Verification of phage lysing Serratia marcescens

[0043] 1. Inoculate 1% (i.e., 100ul Serratia marcescens bacteria liquid into 10ml non-antibiotic LB liquid medium) Serratia marcescens in 10ml non-antibiotic liquid LB medium, 37°C incubator, 220r / min shaking culture for 4h.

[0044] 2. Divide the bacterial solution into two parts in the BSL-2 biological safety cabinet, one part continues to be cultivated, and the other part is inserted into the phage at a concentration of 1‰. Incubate statically in a 37°C incubator for 12h, and shake at 220r / min for 4h.

[0045] 3. Prepare an ordinary LB solid plate, divide the plate into 2 areas with a marker pen, absorb 100ul of the above two bacterial solutions and drop them on the center of the plate, spread evenly with a sterile coating stick, and incubate in a 37°C incubator for 12h .

[0046] The result is as figure 2 Shown: Serratia marcescens grows evenly in the area that is coated with the bacterium liquid tha...

Embodiment 3

[0047] The lysing ability identification of embodiment 3 bacteriophage

[0048] 1. Select the female Aedes aegypti that has been eclosion for 5-7 days, and at first feed the mosquitoes with a 10% sucrose solution (20 units of penicillin and 20 mg of streptomycin per milliliter) containing antibiotics for three consecutive days and change every 24 hours.

[0049]2. After three days of feeding, remove the sucrose solution containing antibiotics, and let them starve for 24 hours at a temperature of 28°C and a humidity of 80%.

[0050] 3. Then feed the sucrose solution with Serratia marcescens for three consecutive days.

[0051] 4. The above-mentioned mosquitoes were divided into two groups, wherein one group was only fed with sucrose solution, and the other group was fed with the phage containing the above-mentioned embodiment 1 (1×10 8 PFU / ml) of sucrose solution.

[0052] 5. After 48 hours, 5 mosquitoes were selected from each group for the experiment. First, wash the surfa...

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Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to a phage and application thereof. Serratia marcescens is split by the phage, the phage is preserved in Guangdong Microbial Culture Collection Center, and the preservation number is GDMCC NO.61141. The phage provides an effective scheme for clinical treatment of serratia marcescens infection.

Description

technical field [0001] The application belongs to the technical field of biomedicine, and in particular relates to a bacteriophage and its application. Background technique [0002] Bacteriophages (Phage) are widely distributed in nature and can be found in soil or the intestines of animals. Phages are intracellular parasites with strict host specificity. In recent years, due to the continuous discovery of drug resistance and even multi-drug resistance of pathogenic bacteria in clinical practice, the use of specific phages for alternative therapy is the current research direction. [0003] Serratia marcescens (Serratia marcescens) is widely distributed in nature and exists in large quantities in water, soil, and the environment. It is a common clinical opportunistic pathogen that can cause lung, urinary tract, meningeal, and wound infections. septicemia. Due to the abuse and non-standard use of antibiotics, Serratia marcescens has also developed drug resistance clinically...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K35/76A61P31/04
CPCC12N7/00A61K35/76A61P31/04C12N2795/00021Y02A50/30
Inventor 肖小平张仁利程功
Owner SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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