A molecularly defined skin test reagent for the diagnosis of bovine tuberculosis compatible with vaccination against Johne’s Disease

Abstract Tuberculin Purified Protein Derivatives (PPDs) exhibit multiple limitations: they are crude extracts from mycobacterial cultures with largely unknown active components; their production depends on culture of mycobacteria requiring expensive BCL3 production facilities; and their potency depe...

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Autores principales:	Sonya Middleton, Sabine Steinbach, Michael Coad, Kevina McGill, Colm Brady, Anthony Duignan, Jimmy Wiseman, Eamonn Gormley, Gareth J. Jones, H. Martin Vordermeier
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/658a113a41b84f21bdb60cea64264234
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spelling	oai:doaj.org-article:658a113a41b84f21bdb60cea642642342021-12-02T14:06:50ZA molecularly defined skin test reagent for the diagnosis of bovine tuberculosis compatible with vaccination against Johne’s Disease10.1038/s41598-021-82434-72045-2322https://doaj.org/article/658a113a41b84f21bdb60cea642642342021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-82434-7https://doaj.org/toc/2045-2322Abstract Tuberculin Purified Protein Derivatives (PPDs) exhibit multiple limitations: they are crude extracts from mycobacterial cultures with largely unknown active components; their production depends on culture of mycobacteria requiring expensive BCL3 production facilities; and their potency depends on the technically demanding guinea pig assay. To overcome these limitations, we developed a molecularly defined tuberculin (MDT) by adding further antigens to our prototype reagent composed of ESAT-6, CFP-10 and Rv3615c (DIVA skin test, DST). In vitro screening using PBMC from infected and uninfected cattle shortlisted four antigens from a literature-based list of 18 to formulate the MDT. These four antigens plus the previously identified Rv3020c protein, produced as recombinant proteins or overlapping synthetic peptides, were formulated together with the three DST antigens into the MDT to test cattle experimentally and naturally infected with M. bovis, uninfected cattle and MAP vaccinated calves. We demonstrated significant increases in MDT-induced skin responses compared to DST in infected animals, whilst maintaining high specificity in unvaccinated or MAP vaccinated calves. Further, MDT can also be applied in in vitro blood-based interferon-gamma release assays. Thus, MDT promises to be a robust diagnostic skin and blood test reagent overcoming some of the limitations of PPDs and warrants full validation.Sonya MiddletonSabine SteinbachMichael CoadKevina McGillColm BradyAnthony DuignanJimmy WisemanEamonn GormleyGareth J. JonesH. Martin VordermeierNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sonya Middleton Sabine Steinbach Michael Coad Kevina McGill Colm Brady Anthony Duignan Jimmy Wiseman Eamonn Gormley Gareth J. Jones H. Martin Vordermeier A molecularly defined skin test reagent for the diagnosis of bovine tuberculosis compatible with vaccination against Johne’s Disease
description	Abstract Tuberculin Purified Protein Derivatives (PPDs) exhibit multiple limitations: they are crude extracts from mycobacterial cultures with largely unknown active components; their production depends on culture of mycobacteria requiring expensive BCL3 production facilities; and their potency depends on the technically demanding guinea pig assay. To overcome these limitations, we developed a molecularly defined tuberculin (MDT) by adding further antigens to our prototype reagent composed of ESAT-6, CFP-10 and Rv3615c (DIVA skin test, DST). In vitro screening using PBMC from infected and uninfected cattle shortlisted four antigens from a literature-based list of 18 to formulate the MDT. These four antigens plus the previously identified Rv3020c protein, produced as recombinant proteins or overlapping synthetic peptides, were formulated together with the three DST antigens into the MDT to test cattle experimentally and naturally infected with M. bovis, uninfected cattle and MAP vaccinated calves. We demonstrated significant increases in MDT-induced skin responses compared to DST in infected animals, whilst maintaining high specificity in unvaccinated or MAP vaccinated calves. Further, MDT can also be applied in in vitro blood-based interferon-gamma release assays. Thus, MDT promises to be a robust diagnostic skin and blood test reagent overcoming some of the limitations of PPDs and warrants full validation.
format	article
author	Sonya Middleton Sabine Steinbach Michael Coad Kevina McGill Colm Brady Anthony Duignan Jimmy Wiseman Eamonn Gormley Gareth J. Jones H. Martin Vordermeier
author_facet	Sonya Middleton Sabine Steinbach Michael Coad Kevina McGill Colm Brady Anthony Duignan Jimmy Wiseman Eamonn Gormley Gareth J. Jones H. Martin Vordermeier
author_sort	Sonya Middleton
title	A molecularly defined skin test reagent for the diagnosis of bovine tuberculosis compatible with vaccination against Johne’s Disease
title_short	A molecularly defined skin test reagent for the diagnosis of bovine tuberculosis compatible with vaccination against Johne’s Disease
title_full	A molecularly defined skin test reagent for the diagnosis of bovine tuberculosis compatible with vaccination against Johne’s Disease
title_fullStr	A molecularly defined skin test reagent for the diagnosis of bovine tuberculosis compatible with vaccination against Johne’s Disease
title_full_unstemmed	A molecularly defined skin test reagent for the diagnosis of bovine tuberculosis compatible with vaccination against Johne’s Disease
title_sort	molecularly defined skin test reagent for the diagnosis of bovine tuberculosis compatible with vaccination against johne’s disease
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/658a113a41b84f21bdb60cea64264234
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A molecularly defined skin test reagent for the diagnosis of bovine tuberculosis compatible with vaccination against Johne’s Disease

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