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Lysin-antibiotic Synergy Determination Assay

Creative Biolabs is a world-leading innovative biotechnology company that has been at the forefront of lysin research for decades. We specialize in providing lysin-antibiotic synergistic assay services to your specific needs. Our vision is to provide effective solutions and one-stop services to help global customers achieve major breakthroughs in the field of lysin research.

Background

In recent years, phage lysins have emerged as promising solutions to address the problem of antibiotic resistance. Compared with antibiotics, lysins possess several characteristics, such as rapid antibacterial activity in vitro and in vivo, strong receptor binding affinity, safety, narrow cleavage spectrum, low likelihood of bacterial resistance, and relatively easy to be modified by genetic engineering. These properties of lysins make them promising antimicrobial candidates against gram-positive pathogens. Notably, lysins can act synergistically with antibiotics by resensitizing bacteria to insensitive antibiotics. Various lysins, both naturally occurring and engineered, have the potential to be developed into antimicrobials that act synergistically with antibiotics.

CF-301 synergizes with antibiotics in vitro.Fig.1 CF-301 synergizes with antibiotics in vitro. (Schuch, 2014)

Advantages of Synergy

Antimicrobials used in combination should be complementary and synergistic. Lysins are ideal for combination use as their desirable properties complement the action of antibiotics. Lysins do not require bacterial metabolism to be active and are bacteriolytic upon contact. Potent anti-biofilm activity is an essential feature of the lysin family, which contributes to the access of antibiotics to bacteria, effectively complementing the action of antibiotics. Furthermore, narrow spectrum action is a beneficial property of lysins. Unlike broad-spectrum antibiotics, lysins do not kill beneficial microbes, which is beneficial for reducing unnecessary clinical complications.

Assays for Lysin-antibiotic Synergy

At Creative Biolabs, various assays are available and can be used to assess synergistic in vitro antibacterial activity. Assays we offer include but are not limited to:

  • Disk agar diffusion assay

Synergy testing based on this approach provides qualitative screening for interactions between different antimicrobial agents. The steps of this method are relatively simple. First, the agar plates are inoculated with the test microorganisms. Then, filter paper discs containing the desired concentration of the test compound are placed on the agar surface. Petri dishes are grown under appropriate conditions and measurements and statistics were performed. Compared with other methods, the disc diffusion method has many advantages, such as low cost, simple operation, ability to test a large number of microorganisms and antibacterial agents, and easy analysis of the results.

Agar diffusion methods.Fig.2 Agar diffusion methods. (Balouiri, 2016)

  • Broth microdilution checkerboard assay

This assay is frequently used to determine interactions among antimicrobial agents. It is a two-dimensional method for evaluating the bactericidal activity of specific concentration combinations of antimicrobial agents. In vitro interactions are calculated and interpreted as synergistic, additive, unrelated, or antagonistic by comparing the antibacterial activity of the agents in combination with those applied individually.

  • Time-kill assay

The time-kill test is the most appropriate method to determine the bactericidal effect, which reveals a time-dependent or concentration-dependent antimicrobial effect. This method is important for determining synergy or antagonism between combined drugs. Furthermore, it is closely linked to obtaining information on the dynamic interactions between the combined drugs and strains.

With extensive experience and advanced platforms, Creative Biolabs provides comprehensive lysin-antibiotic synergy assay services, including but not limited to disk agar diffusion assay, time-kill assay, and broth microdilution checkerboard assay. Please feel free to contact us to discuss your project plans.

References:

  1. Schuch, R.; et al. Combination therapy with lysin CF-301 and antibiotic is superior to antibiotic alone for treating methicillin-resistant Staphylococcus aureus-induced murine bacteremia. The Journal of infectious diseases. 2014, 209(9): 1469-1478.
  2. Balouiri, M.; et al. Methods for in vitro evaluating antimicrobial activity: A review. Journal of pharmaceutical analysis. 2016, 6(2): 71-79.
For Research Use Only. Do NOT use in humans.

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