Separation and screening method based on Listeria monocytogenes bacteriophage
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A Listeria monocytogenes screening method technology, applied in the direction of microorganism-based methods, viruses/bacteriophages, biochemical equipment and methods, etc., can solve the problems of endangering human health, high food residues, enhancing Listeria monocytogenes, etc. problems, to achieve the effect of solving drug resistance, transforming specificity, and improving difficulty
Pending Publication Date: 2021-07-20
GANSU AGRI UNIV
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What deserves our attention most is that there are currently many studies showing that environmental stress can enhance the ability of Listeria monocytogenes to tolerate gastric juice, which seriously endangers human health
[0005] In recent years, with the widespread use of antibiotics, the abuse and high residues in food have caused a large number of food public health incidents, and in the long-term evolution of bacteria, viruses and other pathogens, many antibiotics have shown poor efficacy. Super bacteria and viruses with obvious resistance to antibiotics
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[0056] Below in conjunction with embodiment the present invention is described in further detail:
[0057] The present invention provides a technical solution: based on the separation and screening method of Listeria monocytogenes phage, which is based on the separation of phages from untreated slaughter sewage in pig slaughterhouses, the separation method based on Listeria monocytogenes phages comprises the following steps :
[0058] Step 1. Prepare materials and reagents, including: LB medium, DNase, RNase, 0.22 μm filter membrane, SM buffer, cesium chloride, TSB-YE medium, DNA extraction phenol reagent and PEG8000;
[0059] Step 2. Sample processing:
[0060] a. Use a centrifuge tube to hold the sewage and temporarily store it in a constant temperature sampling box with ice packs;
[0061] b. Take it to the laboratory for experiments within 12 hours, and store the rest of the samples in a 4°C refrigerator for later use;
[0062] c. Centrifuge the collected samples at low...
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Abstract
The invention discloses a separation and screening method based on Listeria monocytogenes bacteriophage, and relates to the technical field of bioengineering. Bacteriophage separation is carried out according to untreated slaughter sewage of a live pig slaughter house, and the separation method based on the Listeria monocytogenes bacteriophage comprises the following steps: preparing materials and reagents; performing sample treatment; and separating and screening the bacteriophage. The lysing LM bacteriophage is separated from nature and is subjected to morphological observation; the titer, the one-step growth curve and the optimal infection complex number of the bacteriophage are measured, a host spectrum is determined; the structural basis, the phagocytosis ability and the phagocytosis specificity of the lysing LM bacteriophage function are known, and standardized preparation of samples acting on subsequent clinical research is achieved. An optimal preservation method of the bacteriophage is explored, and a data basis is provided for subsequent gene modification, induced mutation and protein expression by extracting genomes for sequencing and researching action genes and structural genes of the bacteriophage.
Description
technical field [0001] The invention relates to a separation and screening method based on Listeria monocytogenes phage, and relates to the technical field of bioengineering. Background technique [0002] Listeria monocytogenes (Listeria monocytogenes, LM) was first discovered by scholars at the University of Cambridge in 1924 in experimental rabbits suffering from sepsis and reported publicly in 1926. The bacterium is a Gram-positive short bacillus, 0.4-0.5×0.5-2 μm, growing singly, in short chains or connected to each other in a V shape, aerobic or facultatively anaerobic, without spores, and does not produce capsules. According to the bacterial (O) antigen and flagellar (H) antigen, Listeria monocytogenes is divided into 13 serotypes: 1 / 2a, 1 / 2b, 1 / 2c, 3a, 3b, 3c, 4a, 4b, 4ab , 4c, 4d, 4e and 7, the most common serotypes of pathogenic strains were 1 / 2a and 4b. [0003] Listeria monocytogenes can contaminate dairy products, meat products, aquatic products, and ready-to-e...
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