Journal
NANOMATERIALS
Volume 12, Issue 13, Pages -Publisher
MDPI
DOI: 10.3390/nano12132164
Keywords
nanoparticles; antimicrobial activity assessment; minimum inhibitory concentration; luminescence; E. coli
Categories
Funding
- Associate Laboratory for Green Chemistry-LAQV - FCT/MCTES [UIDB/50006/2020, UIDP/50006/2020]
- Scientific Society PROTEOMASS (Portugal)
- FCT/MEC (Portugal) [CEECIND/00648/2017, PD/BD/142865/2018]
- FCT-Foundation for Science and Technology, I.P. [UIDB/04077/2020, UIDP/04077/2020]
- Fundação para a Ciência e a Tecnologia [PD/BD/142865/2018] Funding Source: FCT
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The use of nanoparticles in various industries has raised concerns about their toxicity and antimicrobial activity assessment, as well as the development of standardized handling protocols. This paper presents a fast luminescence-based assay for evaluating the antimicrobial activity of nanoparticles, providing a reliable method for accurately determining inhibitory concentrations and reducing background interference.
The use of nanoparticles in multiple industries has raised concerned voices about the assessment of their toxicity/antimicrobial activity and the development of standardized handling protocols. Issues emerge during the antimicrobial assaying of multiple cargo, colorimetric, colloidal nanoformulations, as standard protocols often rely on visual evaluations, or optical density (OD) measurements, leading to high variance inhibitory concentrations (MIC). Thus, a fast, luminescence-based assay for the effective assessment of the antimicrobial activity of nanoparticles is herein reported, using the bioluminescence of an in-house E. coli ATCC (R) 8739 (TM) construct with the pMV306G13 + Lux plasmid (E. coli Lux). The new strain's sensitivity to ofloxacin as a standard antibiotic was confirmed, and the methodology robustness verified against multiple nanoparticles and colorimetric drugs. The reduction of incubation from 24 to only 8 h, and the sole use of luminescence (LUX490) to accurately determine and distinguish MIC50 and MIC90, are two main advantages of the method. By discarding OD measurements, one can avoid turbidity and color interferences when calculating bacterial growth. This approach is an important tool that contributes to the standardization of methods, reducing samples' background interference and focusing on luminescence as a direct probe for bacterial metabolic activity, growth and, most importantly, the correct assessment of nanomaterials' antimicrobial activity.
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