Prion Disease Blood Test Using Immunoprecipitation and Improved Quaking-Induced Conversion

ABSTRACT A key challenge in managing transmissible spongiform encephalopathies (TSEs) or prion diseases in medicine, agriculture, and wildlife biology is the development of practical tests for prions that are at or below infectious levels. Of particular interest are tests capable of detecting prions...

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Autores principales:	Christina D. Orrú, Jason M. Wilham, Lynne D. Raymond, Franziska Kuhn, Björn Schroeder, Alex J. Raeber, Byron Caughey
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2011
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/7094531be417413aa325dd4da1144e77
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spelling	oai:doaj.org-article:7094531be417413aa325dd4da1144e772021-11-15T15:38:49ZPrion Disease Blood Test Using Immunoprecipitation and Improved Quaking-Induced Conversion10.1128/mBio.00078-112150-7511https://doaj.org/article/7094531be417413aa325dd4da1144e772011-07-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00078-11https://doaj.org/toc/2150-7511ABSTRACT A key challenge in managing transmissible spongiform encephalopathies (TSEs) or prion diseases in medicine, agriculture, and wildlife biology is the development of practical tests for prions that are at or below infectious levels. Of particular interest are tests capable of detecting prions in blood components such as plasma, but blood typically has extremely low prion concentrations and contains inhibitors of the most sensitive prion tests. One of the latter tests is quaking-induced conversion (QuIC), which can be as sensitive as in vivo bioassays, but much more rapid, higher throughput, and less expensive. Now we have integrated antibody 15B3-based immunoprecipitation with QuIC reactions to increase sensitivity and isolate prions from inhibitors such as those in plasma samples. Coupling of immunoprecipitation and an improved real-time QuIC reaction dramatically enhanced detection of variant Creutzfeldt-Jakob disease (vCJD) brain tissue diluted into human plasma. Dilutions of 1014-fold, containing ~2 attogram (ag) per ml of proteinase K-resistant prion protein, were readily detected, indicating ~10,000-fold greater sensitivity for vCJD brain than has previously been reported. We also discriminated between plasma and serum samples from scrapie-infected and uninfected hamsters, even in early preclinical stages. This combined assay, which we call “enhanced QuIC” (eQuIC), markedly improves prospects for routine detection of low levels of prions in tissues, fluids, or environmental samples. IMPORTANCE Transmissible spongiform encephalopathies (TSEs) are largely untreatable and are difficult to diagnose definitively prior to irreversible clinical decline or death. The transmissibility of TSEs within and between species highlights the need for practical tests for even the smallest amounts of infectivity. A few sufficiently sensitive in vitro methods have been reported, but most have major limitations that would preclude their use in routine diagnostic or screening applications. Our new assay improves the outlook for such critical applications. We focused initially on blood plasma because a practical blood test for prions would be especially valuable for TSE diagnostics and risk reduction. Variant Creutzfeldt-Jakob disease (vCJD) in particular has been transmitted between humans via blood transfusions. Enhanced real-time quaking-induced conversion (eRTQ) provides by far the most sensitive detection of vCJD to date. The 15B3 antibody binds prions of multiple species, suggesting that our assay may be useful for clinical and fundamental studies of a variety of TSEs of humans and animals.Christina D. OrrúJason M. WilhamLynne D. RaymondFranziska KuhnBjörn SchroederAlex J. RaeberByron CaugheyAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 2, Iss 3 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Christina D. Orrú Jason M. Wilham Lynne D. Raymond Franziska Kuhn Björn Schroeder Alex J. Raeber Byron Caughey Prion Disease Blood Test Using Immunoprecipitation and Improved Quaking-Induced Conversion
description	ABSTRACT A key challenge in managing transmissible spongiform encephalopathies (TSEs) or prion diseases in medicine, agriculture, and wildlife biology is the development of practical tests for prions that are at or below infectious levels. Of particular interest are tests capable of detecting prions in blood components such as plasma, but blood typically has extremely low prion concentrations and contains inhibitors of the most sensitive prion tests. One of the latter tests is quaking-induced conversion (QuIC), which can be as sensitive as in vivo bioassays, but much more rapid, higher throughput, and less expensive. Now we have integrated antibody 15B3-based immunoprecipitation with QuIC reactions to increase sensitivity and isolate prions from inhibitors such as those in plasma samples. Coupling of immunoprecipitation and an improved real-time QuIC reaction dramatically enhanced detection of variant Creutzfeldt-Jakob disease (vCJD) brain tissue diluted into human plasma. Dilutions of 1014-fold, containing ~2 attogram (ag) per ml of proteinase K-resistant prion protein, were readily detected, indicating ~10,000-fold greater sensitivity for vCJD brain than has previously been reported. We also discriminated between plasma and serum samples from scrapie-infected and uninfected hamsters, even in early preclinical stages. This combined assay, which we call “enhanced QuIC” (eQuIC), markedly improves prospects for routine detection of low levels of prions in tissues, fluids, or environmental samples. IMPORTANCE Transmissible spongiform encephalopathies (TSEs) are largely untreatable and are difficult to diagnose definitively prior to irreversible clinical decline or death. The transmissibility of TSEs within and between species highlights the need for practical tests for even the smallest amounts of infectivity. A few sufficiently sensitive in vitro methods have been reported, but most have major limitations that would preclude their use in routine diagnostic or screening applications. Our new assay improves the outlook for such critical applications. We focused initially on blood plasma because a practical blood test for prions would be especially valuable for TSE diagnostics and risk reduction. Variant Creutzfeldt-Jakob disease (vCJD) in particular has been transmitted between humans via blood transfusions. Enhanced real-time quaking-induced conversion (eRTQ) provides by far the most sensitive detection of vCJD to date. The 15B3 antibody binds prions of multiple species, suggesting that our assay may be useful for clinical and fundamental studies of a variety of TSEs of humans and animals.
format	article
author	Christina D. Orrú Jason M. Wilham Lynne D. Raymond Franziska Kuhn Björn Schroeder Alex J. Raeber Byron Caughey
author_facet	Christina D. Orrú Jason M. Wilham Lynne D. Raymond Franziska Kuhn Björn Schroeder Alex J. Raeber Byron Caughey
author_sort	Christina D. Orrú
title	Prion Disease Blood Test Using Immunoprecipitation and Improved Quaking-Induced Conversion
title_short	Prion Disease Blood Test Using Immunoprecipitation and Improved Quaking-Induced Conversion
title_full	Prion Disease Blood Test Using Immunoprecipitation and Improved Quaking-Induced Conversion
title_fullStr	Prion Disease Blood Test Using Immunoprecipitation and Improved Quaking-Induced Conversion
title_full_unstemmed	Prion Disease Blood Test Using Immunoprecipitation and Improved Quaking-Induced Conversion
title_sort	prion disease blood test using immunoprecipitation and improved quaking-induced conversion
publisher	American Society for Microbiology
publishDate	2011
url	https://doaj.org/article/7094531be417413aa325dd4da1144e77
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Prion Disease Blood Test Using Immunoprecipitation and Improved Quaking-Induced Conversion

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