Development of immunohistochemistry and in situ hybridisation for the detection of SARS-CoV and SARS-CoV-2 in formalin-fixed paraffin-embedded specimens

Abstract The rapid emergence of SARS-CoV-2, the causative agent of COVID-19, and its dissemination globally has caused an unprecedented strain on public health. Animal models are urgently being developed for SARS-CoV-2 to aid rational design of vaccines and therapeutics. Immunohistochemistry and in...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Fabian Z. X. Lean, Mart M. Lamers, Samuel P. Smith, Rebecca Shipley, Debby Schipper, Nigel Temperton, Bart L. Haagmans, Ashley C. Banyard, Kevin R. Bewley, Miles W. Carroll, Sharon M. Brookes, Ian Brown, Alejandro Nuñez
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ef9e8d2251f54d968b2d6cfdc2c81861
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:ef9e8d2251f54d968b2d6cfdc2c81861
record_format	dspace
spelling	oai:doaj.org-article:ef9e8d2251f54d968b2d6cfdc2c818612021-12-02T12:42:18ZDevelopment of immunohistochemistry and in situ hybridisation for the detection of SARS-CoV and SARS-CoV-2 in formalin-fixed paraffin-embedded specimens10.1038/s41598-020-78949-02045-2322https://doaj.org/article/ef9e8d2251f54d968b2d6cfdc2c818612020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-78949-0https://doaj.org/toc/2045-2322Abstract The rapid emergence of SARS-CoV-2, the causative agent of COVID-19, and its dissemination globally has caused an unprecedented strain on public health. Animal models are urgently being developed for SARS-CoV-2 to aid rational design of vaccines and therapeutics. Immunohistochemistry and in situ hybridisation techniques that facilitate reliable and reproducible detection of SARS-CoV and SARS-CoV-2 viral products in formalin-fixed paraffin-embedded (FFPE) specimens would be of great utility. A selection of commercial antibodies generated against SARS-CoV spike protein and nucleoprotein, double stranded RNA, and RNA probe for spike genes were evaluated for the ability to detect FFPE infected cells. We also tested both heat- and enzymatic-mediated virus antigen retrieval methods to determine the optimal virus antigen recovery as well as identifying alternative retrieval methods to enable flexibility of IHC methods. In addition to using native virus infected cells as positive control material, the evaluation of non-infected cells expressing coronavirus (SARS, MERS) spike as a biosecure alternative to assays involving live virus was undertaken. Optimized protocols were successfully applied to experimental animal-derived tissues. The diverse techniques for virus detection and control material generation demonstrated in this study can be applied to investigations of coronavirus pathogenesis and therapeutic research in animal models.Fabian Z. X. LeanMart M. LamersSamuel P. SmithRebecca ShipleyDebby SchipperNigel TempertonBart L. HaagmansAshley C. BanyardKevin R. BewleyMiles W. CarrollSharon M. BrookesIan BrownAlejandro NuñezNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-9 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Fabian Z. X. Lean Mart M. Lamers Samuel P. Smith Rebecca Shipley Debby Schipper Nigel Temperton Bart L. Haagmans Ashley C. Banyard Kevin R. Bewley Miles W. Carroll Sharon M. Brookes Ian Brown Alejandro Nuñez Development of immunohistochemistry and in situ hybridisation for the detection of SARS-CoV and SARS-CoV-2 in formalin-fixed paraffin-embedded specimens
description	Abstract The rapid emergence of SARS-CoV-2, the causative agent of COVID-19, and its dissemination globally has caused an unprecedented strain on public health. Animal models are urgently being developed for SARS-CoV-2 to aid rational design of vaccines and therapeutics. Immunohistochemistry and in situ hybridisation techniques that facilitate reliable and reproducible detection of SARS-CoV and SARS-CoV-2 viral products in formalin-fixed paraffin-embedded (FFPE) specimens would be of great utility. A selection of commercial antibodies generated against SARS-CoV spike protein and nucleoprotein, double stranded RNA, and RNA probe for spike genes were evaluated for the ability to detect FFPE infected cells. We also tested both heat- and enzymatic-mediated virus antigen retrieval methods to determine the optimal virus antigen recovery as well as identifying alternative retrieval methods to enable flexibility of IHC methods. In addition to using native virus infected cells as positive control material, the evaluation of non-infected cells expressing coronavirus (SARS, MERS) spike as a biosecure alternative to assays involving live virus was undertaken. Optimized protocols were successfully applied to experimental animal-derived tissues. The diverse techniques for virus detection and control material generation demonstrated in this study can be applied to investigations of coronavirus pathogenesis and therapeutic research in animal models.
format	article
author	Fabian Z. X. Lean Mart M. Lamers Samuel P. Smith Rebecca Shipley Debby Schipper Nigel Temperton Bart L. Haagmans Ashley C. Banyard Kevin R. Bewley Miles W. Carroll Sharon M. Brookes Ian Brown Alejandro Nuñez
author_facet	Fabian Z. X. Lean Mart M. Lamers Samuel P. Smith Rebecca Shipley Debby Schipper Nigel Temperton Bart L. Haagmans Ashley C. Banyard Kevin R. Bewley Miles W. Carroll Sharon M. Brookes Ian Brown Alejandro Nuñez
author_sort	Fabian Z. X. Lean
title	Development of immunohistochemistry and in situ hybridisation for the detection of SARS-CoV and SARS-CoV-2 in formalin-fixed paraffin-embedded specimens
title_short	Development of immunohistochemistry and in situ hybridisation for the detection of SARS-CoV and SARS-CoV-2 in formalin-fixed paraffin-embedded specimens
title_full	Development of immunohistochemistry and in situ hybridisation for the detection of SARS-CoV and SARS-CoV-2 in formalin-fixed paraffin-embedded specimens
title_fullStr	Development of immunohistochemistry and in situ hybridisation for the detection of SARS-CoV and SARS-CoV-2 in formalin-fixed paraffin-embedded specimens
title_full_unstemmed	Development of immunohistochemistry and in situ hybridisation for the detection of SARS-CoV and SARS-CoV-2 in formalin-fixed paraffin-embedded specimens
title_sort	development of immunohistochemistry and in situ hybridisation for the detection of sars-cov and sars-cov-2 in formalin-fixed paraffin-embedded specimens
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/ef9e8d2251f54d968b2d6cfdc2c81861
work_keys_str_mv	AT fabianzxlean developmentofimmunohistochemistryandinsituhybridisationforthedetectionofsarscovandsarscov2informalinfixedparaffinembeddedspecimens AT martmlamers developmentofimmunohistochemistryandinsituhybridisationforthedetectionofsarscovandsarscov2informalinfixedparaffinembeddedspecimens AT samuelpsmith developmentofimmunohistochemistryandinsituhybridisationforthedetectionofsarscovandsarscov2informalinfixedparaffinembeddedspecimens AT rebeccashipley developmentofimmunohistochemistryandinsituhybridisationforthedetectionofsarscovandsarscov2informalinfixedparaffinembeddedspecimens AT debbyschipper developmentofimmunohistochemistryandinsituhybridisationforthedetectionofsarscovandsarscov2informalinfixedparaffinembeddedspecimens AT nigeltemperton developmentofimmunohistochemistryandinsituhybridisationforthedetectionofsarscovandsarscov2informalinfixedparaffinembeddedspecimens AT bartlhaagmans developmentofimmunohistochemistryandinsituhybridisationforthedetectionofsarscovandsarscov2informalinfixedparaffinembeddedspecimens AT ashleycbanyard developmentofimmunohistochemistryandinsituhybridisationforthedetectionofsarscovandsarscov2informalinfixedparaffinembeddedspecimens AT kevinrbewley developmentofimmunohistochemistryandinsituhybridisationforthedetectionofsarscovandsarscov2informalinfixedparaffinembeddedspecimens AT mileswcarroll developmentofimmunohistochemistryandinsituhybridisationforthedetectionofsarscovandsarscov2informalinfixedparaffinembeddedspecimens AT sharonmbrookes developmentofimmunohistochemistryandinsituhybridisationforthedetectionofsarscovandsarscov2informalinfixedparaffinembeddedspecimens AT ianbrown developmentofimmunohistochemistryandinsituhybridisationforthedetectionofsarscovandsarscov2informalinfixedparaffinembeddedspecimens AT alejandronunez developmentofimmunohistochemistryandinsituhybridisationforthedetectionofsarscovandsarscov2informalinfixedparaffinembeddedspecimens
_version_	1718393717341028352

Development of immunohistochemistry and in situ hybridisation for the detection of SARS-CoV and SARS-CoV-2 in formalin-fixed paraffin-embedded specimens

Ejemplares similares