Cargando…
Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli
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 nanoformulation...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9268724/ https://www.ncbi.nlm.nih.gov/pubmed/35807997 http://dx.doi.org/10.3390/nano12132164 |
_version_ | 1784744055249305600 |
---|---|
author | Marcelo, Gonçalo A. Galhano, Joana Duarte, Maria Paula Capelo-Martínez, José Luis Lodeiro, Carlos Oliveira, Elisabete |
author_facet | Marcelo, Gonçalo A. Galhano, Joana Duarte, Maria Paula Capelo-Martínez, José Luis Lodeiro, Carlos Oliveira, Elisabete |
author_sort | Marcelo, Gonçalo A. |
collection | PubMed |
description | 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(®) 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 (LUX(490)) to accurately determine and distinguish MIC(50) and MIC(90), 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. |
format | Online Article Text |
id | pubmed-9268724 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-92687242022-07-09 Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli Marcelo, Gonçalo A. Galhano, Joana Duarte, Maria Paula Capelo-Martínez, José Luis Lodeiro, Carlos Oliveira, Elisabete Nanomaterials (Basel) Article 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(®) 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 (LUX(490)) to accurately determine and distinguish MIC(50) and MIC(90), 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. MDPI 2022-06-23 /pmc/articles/PMC9268724/ /pubmed/35807997 http://dx.doi.org/10.3390/nano12132164 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Marcelo, Gonçalo A. Galhano, Joana Duarte, Maria Paula Capelo-Martínez, José Luis Lodeiro, Carlos Oliveira, Elisabete Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli |
title | Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli |
title_full | Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli |
title_fullStr | Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli |
title_full_unstemmed | Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli |
title_short | Validation of a Standard Luminescence Method for the Fast Determination of the Antimicrobial Activity of Nanoparticles in Escherichia coli |
title_sort | validation of a standard luminescence method for the fast determination of the antimicrobial activity of nanoparticles in escherichia coli |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9268724/ https://www.ncbi.nlm.nih.gov/pubmed/35807997 http://dx.doi.org/10.3390/nano12132164 |
work_keys_str_mv | AT marcelogoncaloa validationofastandardluminescencemethodforthefastdeterminationoftheantimicrobialactivityofnanoparticlesinescherichiacoli AT galhanojoana validationofastandardluminescencemethodforthefastdeterminationoftheantimicrobialactivityofnanoparticlesinescherichiacoli AT duartemariapaula validationofastandardluminescencemethodforthefastdeterminationoftheantimicrobialactivityofnanoparticlesinescherichiacoli AT capelomartinezjoseluis validationofastandardluminescencemethodforthefastdeterminationoftheantimicrobialactivityofnanoparticlesinescherichiacoli AT lodeirocarlos validationofastandardluminescencemethodforthefastdeterminationoftheantimicrobialactivityofnanoparticlesinescherichiacoli AT oliveiraelisabete validationofastandardluminescencemethodforthefastdeterminationoftheantimicrobialactivityofnanoparticlesinescherichiacoli |