Staphylococcal bacteriophage composition and application thereof in medicaments for staphylococcal infection diseases

A staphylococcus and phage technology, applied in the field of microorganisms, can solve the problems of limiting the application of phages, narrow lysis spectrum, and few staphylococcal phages, and achieve the effects of broadening the scope of sterilization, not easy to inactivate, improving physical fitness and disease resistance

Active Publication Date: 2020-06-09
QINGDAO PHAGEPHARM BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, few staphylococcal phages have been found so far, and their lytic spectrum is narrow, w

Method used

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  • Staphylococcal bacteriophage composition and application thereof in medicaments for staphylococcal infection diseases
  • Staphylococcal bacteriophage composition and application thereof in medicaments for staphylococcal infection diseases
  • Staphylococcal bacteriophage composition and application thereof in medicaments for staphylococcal infection diseases

Examples

Experimental program
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Effect test

Embodiment 1

[0041] The isolation, purification and amplification of embodiment 1 phage

[0042] Take fecal sewage from a dairy farm in Qingdao City, Henan Province and Shandong Province, add Staphylococcus logarithmic phase bacteria solution, and add appropriate amount of nutrient broth, and place it in a shaking table at 37°C and 170rpm for 16h. 5ml of the liquid was taken out, centrifuged at 11000rpm / min for 10min, and the supernatant was filtered through a 0.22μm sterile microporous membrane to obtain the phage stock solution.

[0043] Isolate phages by using the double-layer plate method, dilute the phage stock solution 10 times, take 100μl of the diluted solution and mix it with 100μl of the corresponding staphylococcus bacteria solution, incubate at 37°C for 5min, mix into 0.8% upper layer agar medium, spread evenly on 1.5 % lower layer agar medium, cooled and incubated overnight in a 37°C incubator. The next day, observe the plaque formation, pick a single phage plaque, repeat the...

Embodiment 2

[0046] Morphological observation of embodiment 2 phage

[0047] Take 20μl and the titer is 1×10 9 The PFU / ml phage suspension was dropped on the copper grid, stained with 2% phosphotungstic acid for 5-10 minutes, put the copper grid on dry filter paper, dried naturally, and observed with Hitachi HT7700 transmission electron microscope.

[0048] Such as image 3 As shown, the head of phage vB-SauM-PB67 has an icosahedral structure, with a head length of 100-110 nm, a head width of 80-90 nm, and a tail length of 200 nm. According to the definition of the International Committee on Taxonomy of Viruses (ICTV), phage vB-SauM- The morphology of PB67 conforms to the characteristics of Myocaudidae and belongs to Myocaudidae.

[0049] Such as Figure 4 As shown, the head of phage vB-SauM-PB93 has an icosahedral structure, the length of the head is 60-70nm, the width of the head is 40-50nm, and the length of the tail is 250nm. The morphology of vB-SauM-PB93 conforms to the character...

Embodiment 3

[0050] The mensuration of embodiment 3 phage lysis spectrum

[0051] The cleavage profile was measured by the double-layer plate method, and single colonies of 102 strains of bovine Staphylococcus aureus isolated from cow mastitis cases and 1 strain of Staphylococcus aureus standard strain (25923) were picked and inoculated in 5ml nutrient broth respectively, and shaken The logarithmic phase bacterial liquid was obtained by culturing in medium. Take 100 μl of bacterial liquid and 100 μl of bacteriophage to lay a double-layer plate according to the conventional method in Example 1. After solidification, incubate overnight in a 37° C. incubator, observe and record the results. Among them, 102 strains of bovine Staphylococcus aureus come from various regions in Shandong, namely Laixi, Rizhao, etc., and 1 strain is the standard strain of Staphylococcus aureus (25923); among the 103 strains of Staphylococcus aureus, 90 strains are methoxygen-resistant Staphylococcus aureus (MRSA),...

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Abstract

The invention discloses a staphylococcal bacteriophage composition and an application thereof in medicaments for staphylococcal infection diseases. The bacteriophage composition is formed by compounding a staphylococcal bacteriophage vB-SauM-PB67 strain and a staphylococcal bacteriophage vB-SauM-PB93, and also discloses a pharmaceutical composition, feed additive and disinfectant taking the bacteriophage composition as a main active component and an application of the pharmaceutical composition. The bacteriophage composition has a strong lysis effect on bovine staphylococcus and can be used for preventing and treating diseases caused by bovine staphylococcus infection. Compared with single lysis single-strain bacteriophage, the bacteriophage composition greatly widens the sterilization range, increases sterilization activity, and can improve physique and disease resistance of dairy cows. Moreover, the bacteriophage composition is low in cost and is environmentally friendly.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a staphylococcus phage composition and its application in medicines for preventing and treating staphylococcus infection diseases. Background technique [0002] Staphylococcus is a Gram-positive bacterium with a diameter of about 0.8 μm and a small spherical shape that can decompose mannitol. Most of the strains are positive for coagulase and catalase. Staphylococcus is the main pathogen of dairy cow mastitis. It is widely distributed in nature, cow teats and the skin surface of cows, and can be spread in cows through milking and other methods. With the wide application of antibiotics, it shows more and more common multi-drug resistance, which is very easy to bury safety hazards to the health of dairy cows and the hygiene and safety of milk and other foods. [0003] As a kind of virus, bacteriophage is a new type of biological agent with strong specificity, high safety an...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61P31/04A61P15/14A23K10/18A01N63/40A01P1/00C12R1/92
CPCC12N7/00A61K35/76A61P31/04A61P15/14A23K10/18A01N63/00C12N2795/10121C12N2795/10131C12N2795/10132
Inventor 潘强任慧英孙虎芝闫艳新徐帆
Owner QINGDAO PHAGEPHARM BIO TECH CO LTD
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