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Rapid detection of φX-174 virus based on synchronous fluorescence of tryptophan

The development of rapid methods for the detection of virus particles based on their intrinsic fluorescence (the native auto-fluorescence that originates from the non-labeled analyte) is challenging. Pure viruses may be detected in filtered solutions, based on the strong fluorescence of the amino ac...

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Autores principales: Farber, Yair, Shlosberg, Yaniv, Schechter, Israel, Armon, Robert
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9702944/
https://www.ncbi.nlm.nih.gov/pubmed/36441232
http://dx.doi.org/10.1007/s00216-022-04436-2
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author Farber, Yair
Shlosberg, Yaniv
Schechter, Israel
Armon, Robert
author_facet Farber, Yair
Shlosberg, Yaniv
Schechter, Israel
Armon, Robert
author_sort Farber, Yair
collection PubMed
description The development of rapid methods for the detection of virus particles based on their intrinsic fluorescence (the native auto-fluorescence that originates from the non-labeled analyte) is challenging. Pure viruses may be detected in filtered solutions, based on the strong fluorescence of the amino acid tryptophan (Trp) in their proteins. Nevertheless, Trp also exists in high quantities in the hosts and host cultivation media. In this work, we developed a new method for the detection of the naked φX-174 virus. We show that a separation of φX-174 from its Escherichia coli host (grown on the standard cultivation medium nutrient agar) by simple extraction and filtration is not sufficient for its detection based on the intrinsic fluorescence since ~ 70% of the Trp fluorescence is derived from impurities. We formulate a new cultivation medium with a very low Trp concentration. We apply synchronous fluorescence measurements to show that no Trp fluorescence is detected in the extract solution upon incubation of this medium substrate with ammonium acetate extraction buffer. Finally, we apply synchronous fluorescence to detect φX-174 based on the spectral fingerprint of its native Trp content. Such a method is more rapid than usual traditional separation and detection methods which can take several hours and does not require any addition of labeling agents such as fluorescent dyes or antibodies for the detection. As other virus species contain Trp as one of the amino acids presents in their proteins, this method has the potential to apply to the detection of other viral species. GRAPHICAL ABSTRACT: [Image: see text]
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spelling pubmed-97029442022-11-28 Rapid detection of φX-174 virus based on synchronous fluorescence of tryptophan Farber, Yair Shlosberg, Yaniv Schechter, Israel Armon, Robert Anal Bioanal Chem Research Paper The development of rapid methods for the detection of virus particles based on their intrinsic fluorescence (the native auto-fluorescence that originates from the non-labeled analyte) is challenging. Pure viruses may be detected in filtered solutions, based on the strong fluorescence of the amino acid tryptophan (Trp) in their proteins. Nevertheless, Trp also exists in high quantities in the hosts and host cultivation media. In this work, we developed a new method for the detection of the naked φX-174 virus. We show that a separation of φX-174 from its Escherichia coli host (grown on the standard cultivation medium nutrient agar) by simple extraction and filtration is not sufficient for its detection based on the intrinsic fluorescence since ~ 70% of the Trp fluorescence is derived from impurities. We formulate a new cultivation medium with a very low Trp concentration. We apply synchronous fluorescence measurements to show that no Trp fluorescence is detected in the extract solution upon incubation of this medium substrate with ammonium acetate extraction buffer. Finally, we apply synchronous fluorescence to detect φX-174 based on the spectral fingerprint of its native Trp content. Such a method is more rapid than usual traditional separation and detection methods which can take several hours and does not require any addition of labeling agents such as fluorescent dyes or antibodies for the detection. As other virus species contain Trp as one of the amino acids presents in their proteins, this method has the potential to apply to the detection of other viral species. GRAPHICAL ABSTRACT: [Image: see text] Springer Berlin Heidelberg 2022-11-28 2023 /pmc/articles/PMC9702944/ /pubmed/36441232 http://dx.doi.org/10.1007/s00216-022-04436-2 Text en © Springer-Verlag GmbH Germany, part of Springer Nature 2022, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Research Paper
Farber, Yair
Shlosberg, Yaniv
Schechter, Israel
Armon, Robert
Rapid detection of φX-174 virus based on synchronous fluorescence of tryptophan
title Rapid detection of φX-174 virus based on synchronous fluorescence of tryptophan
title_full Rapid detection of φX-174 virus based on synchronous fluorescence of tryptophan
title_fullStr Rapid detection of φX-174 virus based on synchronous fluorescence of tryptophan
title_full_unstemmed Rapid detection of φX-174 virus based on synchronous fluorescence of tryptophan
title_short Rapid detection of φX-174 virus based on synchronous fluorescence of tryptophan
title_sort rapid detection of φx-174 virus based on synchronous fluorescence of tryptophan
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9702944/
https://www.ncbi.nlm.nih.gov/pubmed/36441232
http://dx.doi.org/10.1007/s00216-022-04436-2
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