Bacteriostasis method for selectively inhibiting gram-positive bacteria

Abstract

The invention provides a bacteriostasis method for selectively inhibiting gram-positive bacteria. The method uses small molecule PI-n as a bacteriostatic agent, the structure of the bacteriostatic agent is that biimidazole is connected with pyrene fluorophore through 1-4 methylene groups, and the bacteriostatic agent can selectively inhibit the growth of gram-positive bacteria, including staphylococcus aureus, staphylococcus saprophyticus, enterococcus faecalis, bacillus cereus, streptococcus dysgalactiae and the like, and particularly has a certain inhibition effect on clinical methicillin-resistant staphylococcus aureus (MRSA). The action mechanism is that positively charged PI-n molecules can be highly gathered on the surface of gram-positive bacteria, and under the action of a certain dosage of PI-n, the permeability of bacterial cell walls is changed, and bacteria die due to swelling and rupture, so that the bactericidal effect is achieved.

Description

Technical field

[0001] The present invention belongs to the field of biological analysis detection, and in particular to a bacterial method for selectively inhibiting Gram-positive bacteria. Background technique

[0002] Antibiotics is a great discovery in the history of human medicine. It is an important tool for human resistance to infectious diseases caused by pathogenic microbial invasion. Since the first antibiotic-penicillin in the world, various kinds of antibiotics have been discovered and used, such as tetracycline, chloramphenicol, monocyclic β-lactyl ring, and the like. Its action mechanism includes inhibiting the synthesis of bacterial cell wall, changing the permeability of cytospermea, inhibiting the synthesis of proteins, interfering with bacterial DNA replication, etc.. Antibiotics have become an indispensable drug during medical treatment, which is widely used worldwide. However, bacterial adaptation ability is extremely strong, and it can be survived by changing...

Examples

Embodiment 1

[0033] Determination of the minimum inhibitory concentration of PI-1 on different bacterial growth

[0034] From the rapid velococci, Bacillus, Bacillus, Rucfacial Sapoccus, and Sapallococcus S. vectors, respectively, 37 ° C shaker, 37 ° C shake the bed, the second day, the bacterial liquid is transferred. OD in 5 ml lb medium, 37 ° C shaker culture to the bacterial liquid 600 == 0.5, dilution 100 times to get OD 600 = 0.005 bacterial liquid. The 500 μl of bacteria was added to the N [mu] m, and the final concentration was 40 μm, 20 μm, 10 μm, 5 μm, 2.5 μm, 1.25 μm, 0.625 μm, 0.3125 μm, 37 ° C shaker, and observed bacterial growth after 24 hours. For Gram-negative bacteria, the final concentration of Pi-1 is 160 μm, 80 μm, 40 μm, 20 μm, 10 μm, 5 μm, 2.5 μm, 1.25 μm, respectively, 2 μm, 1.25 μm, respectively.

[0035] During the experiment, two sets of parallel experiments were set, and the three sets of controls were set. The control group was: 500 μl of bacterial liquid; 500 μl o...

Embodiment 2

[0040] Study on the inhibition of Wax Nakhococcus Pi-Strans

[0041] 50 μl of the bacterial liquid of 50 μl of wax spurs was added to 5 ml lb medium, and overnight culture was cultured. Take 10 mLLB medium to add bacterial liquid, the initial concentration of the gemmia is OD 600 = 0.01, then subjected to two test tubes. One of the test tubes was added to the PI-2 (final concentration 5 μm), and the other test tube was added to the same volume of DMSO as a control, 37 ° C shaker culture, interval of half an hour genus OD 600 value. Growth by monitoring the growth of wax spurobacillus figure 1 .

[0042] figure 1 In the case of the addition of the same volume DMSO, the wax sprrier is rapidly proliferated, and the bacteria added to PI-2 is substantially non-proliferation, indicating that the addition of PI-2 inhibits the growth of wax sample.

Embodiment 3

[0044]Pi-2 on the inhibition of Bacillus subtilis

[0045] 5 ml lb medium was added 50 μl of Bacillus subtilis, overnight culture. Take 10 mLLB medium to add bacterial liquid, the initial concentration of the gemmia is OD 600 = 0.01, then subjected to two test tubes. One of the test tubes was added to the PI-2 (final concentration 5 μm), and the other test tube was added to the same volume of DMSO as a control, 37 ° C shaker culture, interval of half an hour genus OD 600 value. Growth by monitoring the growth of Bacillus subtilis figure 2 .

[0046] figure 2 Bacillus subtilis rapidly proliferate in the case of adding the same volume DMSO, and the bacteria added to PI-2 substantially does not proliferate, indicating that the addition of Pi-2 inhibits the growth of Bacillus Bacillus.

Technical field

[0001] The present invention belongs to the field of biological analysis detection, and in particular to a bacterial method for selectively inhibiting Gram-positive bacteria. Background technique

[0002] Antibiotics is a great discovery in the history of human medicine. It is an important tool for human resistance to infectious diseases caused by pathogenic microbial invasion. Since the first antibiotic-penicillin in the world, various kinds of antibiotics have been discovered and used, such as tetracycline, chloramphenicol, monocyclic β-lactyl ring, and the like. Its action mechanism includes inhibiting the synthesis of bacterial cell wall, changing the permeability of cytospermea, inhibiting the synthesis of proteins, interfering with bacterial DNA replication, etc.. Antibiotics have become an indispensable drug during medical treatment, which is widely used worldwide. However, bacterial adaptation ability is extremely strong, and it can be survived by changing...

Examples

Embodiment 1

[0033] Determination of the minimum inhibitory concentration of PI-1 on different bacterial growth

[0034] From the rapid velococci, Bacillus, Bacillus, Rucfacial Sapoccus, and Sapallococcus S. vectors, respectively, 37 ° C shaker, 37 ° C shake the bed, the second day, the bacterial liquid is transferred. OD in 5 ml lb medium, 37 ° C shaker culture to the bacterial liquid 600 == 0.5, dilution 100 times to get OD 600 = 0.005 bacterial liquid. The 500 μl of bacteria was added to the N [mu] m, and the final concentration was 40 μm, 20 μm, 10 μm, 5 μm, 2.5 μm, 1.25 μm, 0.625 μm, 0.3125 μm, 37 ° C shaker, and observed bacterial growth after 24 hours. For Gram-negative bacteria, the final concentration of Pi-1 is 160 μm, 80 μm, 40 μm, 20 μm, 10 μm, 5 μm, 2.5 μm, 1.25 μm, respectively, 2 μm, 1.25 μm, respectively.

[0035] During the experiment, two sets of parallel experiments were set, and the three sets of controls were set. The control group was: 500 μl of bacterial liquid; 500 μl o...

Embodiment 2

[0040] Study on the inhibition of Wax Nakhococcus Pi-Strans

[0041] 50 μl of the bacterial liquid of 50 μl of wax spurs was added to 5 ml lb medium, and overnight culture was cultured. Take 10 mLLB medium to add bacterial liquid, the initial concentration of the gemmia is OD 600 = 0.01, then subjected to two test tubes. One of the test tubes was added to the PI-2 (final concentration 5 μm), and the other test tube was added to the same volume of DMSO as a control, 37 ° C shaker culture, interval of half an hour genus OD 600 value. Growth by monitoring the growth of wax spurobacillus figure 1 .

[0042] figure 1 In the case of the addition of the same volume DMSO, the wax sprrier is rapidly proliferated, and the bacteria added to PI-2 is substantially non-proliferation, indicating that the addition of PI-2 inhibits the growth of wax sample.

Embodiment 3

[0044]Pi-2 on the inhibition of Bacillus subtilis

[0045] 5 ml lb medium was added 50 μl of Bacillus subtilis, overnight culture. Take 10 mLLB medium to add bacterial liquid, the initial concentration of the gemmia is OD 600 = 0.01, then subjected to two test tubes. One of the test tubes was added to the PI-2 (final concentration 5 μm), and the other test tube was added to the same volume of DMSO as a control, 37 ° C shaker culture, interval of half an hour genus OD 600 value. Growth by monitoring the growth of Bacillus subtilis figure 2 .

[0046] figure 2 Bacillus subtilis rapidly proliferate in the case of adding the same volume DMSO, and the bacteria added to PI-2 substantially does not proliferate, indicating that the addition of Pi-2 inhibits the growth of Bacillus Bacillus.

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