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Acid-resistant salmonella phage and composition, kit and application thereof

A technology of Salmonella and phage, which is applied in the direction of phage, virus/phage, medical raw materials derived from virus/phage, etc., can solve the problem of weak acid resistance of Salmonella phage, and achieve the effect of no side effects, pollution prevention and strong acid resistance

Active Publication Date: 2020-09-11
PHAGELUX (NANJING) BIO-TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The above-mentioned prior art has the following defects: when the Salmonella enteritidis phage LPSE1 is at pH<3, the phage titer is reduced to 0, indicating that the acid resistance of the Salmonella phage of this bacterial strain is weak

Method used

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  • Acid-resistant salmonella phage and composition, kit and application thereof
  • Acid-resistant salmonella phage and composition, kit and application thereof
  • Acid-resistant salmonella phage and composition, kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1: Screening and purification of phage

[0057] (1) Sample collection and treatment The samples in this example were collected from the sewage of farms in Shandong, Henan and Jiangsu and the soil of nearby farmlands.

[0058] The collected samples were centrifuged at 5000 r / min for 10 min, and passed through a 0.22 μm filter membrane to obtain a filtrate.

[0059] (2) For the enrichment of the target Salmonella phage in the sample, mix the above-mentioned filtrate with 2 times LB liquid medium at a ratio of 1:1, inoculate 100 μL of the target Salmonella strain, and enrich and culture overnight to obtain the enrichment solution.

[0060] (3) Phage screening Centrifuge the above-mentioned enrichment solution to take the supernatant and pass it through a 0.22 μm membrane, then take 1 mL of the filtrate to perform gradient dilution with SM solution, and mix each gradient dilution with 5 mL of LB semi-solid medium containing the target Salmonella , poured on a petri...

Embodiment 2

[0065] Example 2 Determination of Salmonella pullorum phage SG4P1 (Salmonella pullorum phage SG4P1) to the optimal multiplicity of infection (MOI) of Salmonella

[0066]A single colony of Salmonella was picked, inoculated into a test tube containing 3 mL of LB culture solution, and cultured with shaking at 150 rpm at 37°C for 12 hours to obtain a suspension of the host bacteria. The bacterial suspension was transferred to 10 mL LB culture medium at a ratio of 1:100, and cultured at 37°C with shaking at 150 rpm to the pre-logarithmic phase. Add the pure culture solution of Salmonella pullorum phage SG4P1 (prepared by Example 1) and host bacteria (MOI=number of phages / number of bacteria) according to the multiplicity of infection ratio, and add LB liquid medium to make the total volume of each tube the same. Shake overnight at 150 rpm in a shaker at 37°C. After the culture was completed, centrifuge at 5000 g for 10 min and collect the supernatant to measure the phage titer. Ea...

Embodiment 3

[0070] Embodiment 3: the virulence gene of pullorum pullorum phage SG4P1 detects the deletion test of bad gene and selects 65 kinds of virulence genes (table 2) that are derived from lysogenous phage in pathogenic bacteria through identification, by measuring the virulence gene of pullorum pullorum phage SG4P1 The whole genome was analyzed by bioinformatics to determine whether it contained the above-mentioned virulence genes. The results showed that Salmonella pullorum phage SG4P1 did not contain the following virulence genes. The tested phages had no adverse genes.

[0071] Table 2 The main known virulence genes of lysogenic phages in pathogenic bacteria

[0072]

[0073]

[0074] The Salmonella pullorum phage SG4P1 of the present invention does not contain virulence genes or adverse genes, wherein the absence of virulence genes or adverse genes refers to the virulence genes or adverse genes recorded in Table 2.

[0075] Embodiment 3: Toxicology experiment

[0076] ...

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Abstract

The invention belongs to the technical field of salmonella phages, and particularly relates to an acid-resistant phage isolate capable of specifically cracking salmonella and application of the acid-resistant phage isolate in sterilization and bacterium prevention. The invention mainly discloses salmonella pullorum phage SG4P1 with the preservation number of CCTCC NO: M2018765. The salmonella phage is a strong phage isolated from the nature, and toxicological experiments prove that the salmonella phage is safe and free of any side effect. The experiments find that the phage can tolerate extremely high acidity, and therefore, the salmonella phage can have a strong cracking effect on salmonella in the intestinal tract through gastric acid. The phage is expected to be developed into a biological bactericide for preventing and treating infection and pollution of the salmonella.

Description

technical field [0001] The invention belongs to the technical field of Salmonella phage, and in particular relates to an acid-resistant phage capable of specifically lysing Salmonella, a composition, a kit and an application thereof. Background technique [0002] Salmonella generally colonizes the intestines of animals. If it is not handled properly in food processing, it can easily contaminate food, cause food safety hazards, and endanger human health. In recent years, with the abuse of antibiotics by farmers and feed manufacturers, more and more drug-resistant strains of the pathogen have emerged, making many antibiotics for the treatment of Salmonella infection ineffective. According to reports, the resistance of Salmonella in China to sulfonamide antibiotics is close to 100%, and people have to seek other effective prevention and control methods against this bacterial infection. [0003] Phages are a class of viruses that obligately parasitize or infect bacteria, and th...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61P31/04A01N63/40A01P1/00A23K10/18A23K50/75C12R1/92
CPCC12N7/00A61K35/76A61P31/04A01N63/40A23K10/18A23K50/75C12N2795/10121C12N2795/10132C12N2795/10131A61K2300/00
Inventor 费文斌黄杰胡怿林丁良肖逍何四龙陈海刘墨乔欢丛郁谢晓莉徐旭凌
Owner PHAGELUX (NANJING) BIO-TECH CO LTD
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