Development of a sensitive real-time quaking-induced conversion (RT-QuIC) assay for application in prion-infected blood
Efforts to prevent human-to-human transmission of variant Creutzfeldt-Jakob disease (vCJD) by contaminated blood would be aided by the development of a sensitive diagnostic test that could be routinely used to screen blood donations. As blood samples from vCJD patients are extremely rare, here we de...
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| creator | Thomas, Charlotte M Salamat, M. Khalid F de Wolf, Christopher McCutcheon, Sandra Blanco, A. Richard Alejo Manson, Jean C Hunter, Nora Houston, E. Fiona |
| description | Efforts to prevent human-to-human transmission of variant Creutzfeldt-Jakob disease (vCJD) by contaminated blood would be aided by the development of a sensitive diagnostic test that could be routinely used to screen blood donations. As blood samples from vCJD patients are extremely rare, here we describe the optimisation of real-time quaking-induced conversion (RT-QuIC) for detection of PrPSc (misfolded prion protein, a marker of prion infection) in blood samples from an established large animal model of vCJD, sheep experimentally infected with bovine spongiform encephalopathy (BSE). Comparative endpoint titration experiments with RT-QuIC, miniaturized bead protein misfolding cyclic amplification (mb-PMCA) and intracerebral inoculation of a transgenic mouse line expressing sheep PrP (tgOvARQ), demonstrated highly sensitive detection of PrPSc by RT-QuIC in a reference sheep brain homogenate. Upon addition of a capture step with iron oxide beads, the RT-QuIC assay was able to detect PrPSc in whole blood samples from BSE-infected sheep up to two years before disease onset. Both RT-QuIC and mb-PMCA also demonstrated sensitive detection of PrPSc in a reference vCJD-infected human brain homogenate, suggesting that either assay may be suitable for application to human blood samples. Our results support the further development and evaluation of RT-QuIC as a diagnostic or screening test for vCJD. |
| doi_str_mv | 10.1371/journal.pone.0293845 |
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Khalid F ; de Wolf, Christopher ; McCutcheon, Sandra ; Blanco, A. Richard Alejo ; Manson, Jean C ; Hunter, Nora ; Houston, E. Fiona</creator><creatorcontrib>Thomas, Charlotte M ; Salamat, M. Khalid F ; de Wolf, Christopher ; McCutcheon, Sandra ; Blanco, A. Richard Alejo ; Manson, Jean C ; Hunter, Nora ; Houston, E. Fiona</creatorcontrib><description>Efforts to prevent human-to-human transmission of variant Creutzfeldt-Jakob disease (vCJD) by contaminated blood would be aided by the development of a sensitive diagnostic test that could be routinely used to screen blood donations. As blood samples from vCJD patients are extremely rare, here we describe the optimisation of real-time quaking-induced conversion (RT-QuIC) for detection of PrPSc (misfolded prion protein, a marker of prion infection) in blood samples from an established large animal model of vCJD, sheep experimentally infected with bovine spongiform encephalopathy (BSE). Comparative endpoint titration experiments with RT-QuIC, miniaturized bead protein misfolding cyclic amplification (mb-PMCA) and intracerebral inoculation of a transgenic mouse line expressing sheep PrP (tgOvARQ), demonstrated highly sensitive detection of PrPSc by RT-QuIC in a reference sheep brain homogenate. Upon addition of a capture step with iron oxide beads, the RT-QuIC assay was able to detect PrPSc in whole blood samples from BSE-infected sheep up to two years before disease onset. Both RT-QuIC and mb-PMCA also demonstrated sensitive detection of PrPSc in a reference vCJD-infected human brain homogenate, suggesting that either assay may be suitable for application to human blood samples. Our results support the further development and evaluation of RT-QuIC as a diagnostic or screening test for vCJD.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0293845</identifier><language>eng</language><publisher>San Francisco: Public Library of Science</publisher><subject>Animal genetic engineering ; Animal models ; Assaying ; Asymptomatic ; Biology and Life Sciences ; Blood ; Blood & organ donations ; Blood banks ; Blood donors ; Blood tests ; Blood transfusions ; Body fluids ; Bovine spongiform encephalopathy ; Brain ; Brain research ; Care and treatment ; Cattle ; Comparative analysis ; Complications and side effects ; Creutzfeldt-Jakob disease ; Diagnosis ; Disease transmission ; Engineering and Technology ; Ethylenediaminetetraacetic acid ; Evaluation ; Ferric oxide ; Infection ; Infections ; Inoculation ; Iron compounds ; Iron oxides ; Medical screening ; Medical tests ; Medicine and Health Sciences ; Prion diseases ; Prion protein ; Prions ; Protein folding ; Proteins ; Real time ; Research and Analysis Methods ; Research ethics ; Sensitivity analysis ; Sheep ; Titration ; Transgenic mice</subject><ispartof>PloS one, 2023-11, Vol.18 (11), p.e0293845-e0293845</ispartof><rights>COPYRIGHT 2023 Public Library of Science</rights><rights>2023 Thomas et al. 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As blood samples from vCJD patients are extremely rare, here we describe the optimisation of real-time quaking-induced conversion (RT-QuIC) for detection of PrPSc (misfolded prion protein, a marker of prion infection) in blood samples from an established large animal model of vCJD, sheep experimentally infected with bovine spongiform encephalopathy (BSE). Comparative endpoint titration experiments with RT-QuIC, miniaturized bead protein misfolding cyclic amplification (mb-PMCA) and intracerebral inoculation of a transgenic mouse line expressing sheep PrP (tgOvARQ), demonstrated highly sensitive detection of PrPSc by RT-QuIC in a reference sheep brain homogenate. Upon addition of a capture step with iron oxide beads, the RT-QuIC assay was able to detect PrPSc in whole blood samples from BSE-infected sheep up to two years before disease onset. 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Our results support the further development and evaluation of RT-QuIC as a diagnostic or screening test for vCJD.</description><subject>Animal genetic engineering</subject><subject>Animal models</subject><subject>Assaying</subject><subject>Asymptomatic</subject><subject>Biology and Life Sciences</subject><subject>Blood</subject><subject>Blood & organ donations</subject><subject>Blood banks</subject><subject>Blood donors</subject><subject>Blood tests</subject><subject>Blood transfusions</subject><subject>Body fluids</subject><subject>Bovine spongiform encephalopathy</subject><subject>Brain</subject><subject>Brain research</subject><subject>Care and treatment</subject><subject>Cattle</subject><subject>Comparative analysis</subject><subject>Complications and side effects</subject><subject>Creutzfeldt-Jakob disease</subject><subject>Diagnosis</subject><subject>Disease transmission</subject><subject>Engineering and Technology</subject><subject>Ethylenediaminetetraacetic acid</subject><subject>Evaluation</subject><subject>Ferric oxide</subject><subject>Infection</subject><subject>Infections</subject><subject>Inoculation</subject><subject>Iron compounds</subject><subject>Iron oxides</subject><subject>Medical screening</subject><subject>Medical tests</subject><subject>Medicine and Health Sciences</subject><subject>Prion diseases</subject><subject>Prion protein</subject><subject>Prions</subject><subject>Protein folding</subject><subject>Proteins</subject><subject>Real time</subject><subject>Research and Analysis Methods</subject><subject>Research ethics</subject><subject>Sensitivity analysis</subject><subject>Sheep</subject><subject>Titration</subject><subject>Transgenic mice</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNptUk1v1DAQjRBIlMI_QMISl3LI4o_ETk6o2tKyUiUEKmfLccaLF8dO7WRR_z1eNiAWVXOwNfPem8-ieE3wijBB3u_CHL1yqzF4WGHasqaqnxRnpGW05BSzp__8nxcvUtphXLOG87Pi5xXswYVxAD-hYJBCCXyyk90DiqBcOdkB0P2sfli_La3vZw090sHvISYbPLr4eld-mTfrd0ilpB6QCRGpcXRWq-kQtx6NMX8y14CeMrlzIfQvi2dGuQSvlve8-Hb98W79qbz9fLNZX96WuhZkKlmvgCqBa1A1MbxrBCYdB-gEdFhUraFVV1HBWCuEaMCoRjGjcY8NbTKmZufFm6Pu6EKSy6CSZJi3pKVUHBCbI6IPaidzrYOKDzIoK387QtxKFSerHUhOTaew6HpqmipbFmgr3FVQQdNyAlnrw5Jt7gbodR5qVO5E9DTi7Xe5DXtJMKckLyQrXCwKMdzPkCY52KTBOeUhzEnSpuGMclHhDH37H_Tx9hbUVuUO8hJCTqwPovJSCEKJaDHJqNUjqGw9DDZvG4zN_hNCdSToGFKKYP42SbA8HOWfYuThKOVylOwXWVLXgA</recordid><startdate>20231102</startdate><enddate>20231102</enddate><creator>Thomas, Charlotte M</creator><creator>Salamat, M. 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Khalid F</au><au>de Wolf, Christopher</au><au>McCutcheon, Sandra</au><au>Blanco, A. Richard Alejo</au><au>Manson, Jean C</au><au>Hunter, Nora</au><au>Houston, E. Fiona</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of a sensitive real-time quaking-induced conversion (RT-QuIC) assay for application in prion-infected blood</atitle><jtitle>PloS one</jtitle><date>2023-11-02</date><risdate>2023</risdate><volume>18</volume><issue>11</issue><spage>e0293845</spage><epage>e0293845</epage><pages>e0293845-e0293845</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Efforts to prevent human-to-human transmission of variant Creutzfeldt-Jakob disease (vCJD) by contaminated blood would be aided by the development of a sensitive diagnostic test that could be routinely used to screen blood donations. As blood samples from vCJD patients are extremely rare, here we describe the optimisation of real-time quaking-induced conversion (RT-QuIC) for detection of PrPSc (misfolded prion protein, a marker of prion infection) in blood samples from an established large animal model of vCJD, sheep experimentally infected with bovine spongiform encephalopathy (BSE). Comparative endpoint titration experiments with RT-QuIC, miniaturized bead protein misfolding cyclic amplification (mb-PMCA) and intracerebral inoculation of a transgenic mouse line expressing sheep PrP (tgOvARQ), demonstrated highly sensitive detection of PrPSc by RT-QuIC in a reference sheep brain homogenate. Upon addition of a capture step with iron oxide beads, the RT-QuIC assay was able to detect PrPSc in whole blood samples from BSE-infected sheep up to two years before disease onset. Both RT-QuIC and mb-PMCA also demonstrated sensitive detection of PrPSc in a reference vCJD-infected human brain homogenate, suggesting that either assay may be suitable for application to human blood samples. Our results support the further development and evaluation of RT-QuIC as a diagnostic or screening test for vCJD.</abstract><cop>San Francisco</cop><pub>Public Library of Science</pub><doi>10.1371/journal.pone.0293845</doi><orcidid>https://orcid.org/0000-0001-6293-6013</orcidid><orcidid>https://orcid.org/0000-0003-3319-4035</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Animal genetic engineering Animal models Assaying Asymptomatic Biology and Life Sciences Blood Blood & organ donations Blood banks Blood donors Blood tests Blood transfusions Body fluids Bovine spongiform encephalopathy Brain Brain research Care and treatment Cattle Comparative analysis Complications and side effects Creutzfeldt-Jakob disease Diagnosis Disease transmission Engineering and Technology Ethylenediaminetetraacetic acid Evaluation Ferric oxide Infection Infections Inoculation Iron compounds Iron oxides Medical screening Medical tests Medicine and Health Sciences Prion diseases Prion protein Prions Protein folding Proteins Real time Research and Analysis Methods Research ethics Sensitivity analysis Sheep Titration Transgenic mice |
| title | Development of a sensitive real-time quaking-induced conversion (RT-QuIC) assay for application in prion-infected blood |
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