NOTIFy (non-toxic lyophilized field)-FISH for the identification of biological agents by Fluorescence in situ Hybridization.
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2020
2020
Historique:
received:
20
11
2019
accepted:
20
02
2020
entrez:
7
3
2020
pubmed:
7
3
2020
medline:
23
6
2020
Statut:
epublish
Résumé
The rapid and reliable diagnostics of highly pathogenic bacteria under restricted field conditions poses one of the major challenges to medical biodefense, especially since false positive or false negative reports might have far-reaching consequences. Fluorescence in situ hybridization (FISH) has the potential to represent a powerful microscopy-based addition to the existing molecular-based diagnostic toolbox. In this study, we developed a set of FISH-probes for the fast, matrix independent and simultaneous detection of thirteen highly pathogenic bacteria in different environmental and clinical sample matrices. Furthermore, we substituted formamide, a routinely used chemical that is toxic and volatile, by non-toxic urea. This will facilitate the application of FISH under resource limited field laboratory conditions. We demonstrate that hybridizations performed with urea show the same specificity and comparable signal intensities for the FISH-probes used in this study. To further simplify the use of FISH in the field, we lyophilized the reagents needed for FISH. The signal intensities obtained with these lyophilized reagents are comparable to freshly prepared reagents even after storage for a month at room temperature. Finally, we show that by the use of non-toxic lyophilized field (NOTIFy)-FISH, specific detection of microorganisms with simple and easily transportable equipment is possible in the field.
Identifiants
pubmed: 32142548
doi: 10.1371/journal.pone.0230057
pii: PONE-D-19-32120
pmc: PMC7059943
doi:
Substances chimiques
Biological Warfare Agents
0
Formamides
0
formamide
4781T907ZS
Urea
8W8T17847W
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0230057Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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