TY - JOUR KW - bovine spongiform encephalopathy KW - infectivity KW - PrPBSE KW - Protein Misfolding Cyclic Amplification KW - replacement of animal experiments AU - I Ackermann AU - J Shawulu AU - M Keller AU - I Fatola AU - M Groschup AU - A Balkema-Buschmann AB - Classical bovine spongiform encephalopathy (C-BSE) belongs to the transmissible spongiform encephalopathies (TSE), which are also designated prion diseases since they are caused by the conversion of the host-encoded cellular prion protein PrPC to its pathological isoform PrPTSE. BSE carries a zoonotic potential as BSE prions cause variant Creutzfeldt-Jakob disease in humans. To date, C-BSE infectivity can only be detected by bioassay, e.g. highly sensitive bovine PrP transgenic mice (e.g. Tgbov XV mice). Recently, highly sensitive in-vitro prion seeding activity assays, such as the Protein Misfolding Cyclic Amplification (PMCA), have been developed, which work particularly well for the template-assisted prion conversion of scrapie prions, while a similarly efficient bovine C-BSE-prion amplification remained unavailable. In the here described study, we have therefore compared the analytical sensitivities of the transgenic Tgbov XV mouse bioassay and our C-BSE PMCA protocol by analysing serial dilutions of a BSE-positive bovine brainstem homogenate pool. As both methods were shown to possess comparable sensitivities, we propose the C-BSE PMCA as a potential in-vitro replacement method, allowing the reduction and refinement of mouse bioassays for the detection of cattle derived classical BSE prions by reducing them to only specific analytical applications. BT - Berliner und Münchener Tierärztliche Wochenschrift C1 - {"oldId":108497,"title":"Exploring PMCA as a potential in-vitro alternative method to mouse bioassays for the highly sensitive detection of BSE prions","topline":"","teaserText":"Evaluierung der PMCA als In-vitro-Ersatzmethode zum Maus-Bioassay zur hochsensitiven Detektion von BSE-Prionen","content":"

Summary<\/span>
Classical bovine spongiform encephalopathy (C-BSE) belongs to the transmissible spongiform encephalopathies (TSE), which are also designated prion diseases since they are caused by the conversion of the host-encoded cellular prion protein PrPC<\/span> to its pathological isoform PrPTSE<\/span>. BSE carries a zoonotic potential as BSE prions cause variant Creutzfeldt-Jakob disease in humans. To date, C-BSE infectivity can only be detected by bioassay, e.g. highly sensitive bovine PrP transgenic mice (e.g. Tgbov XV mice). Recently, highly sensitive in-vitro prion seeding activity assays, such as the Protein Misfolding Cyclic Amplification (PMCA), have been developed, which work particularly well for the template-assisted prion conversion of scrapie prions, while a similarly efficient bovine C-BSE-prion amplification remained unavailable. In the here described study, we have therefore compared the analytical sensitivities of the transgenic Tgbov XV mouse bioassay and our C-BSE PMCA protocol by analysing serial dilutions of a BSE-positive bovine brainstem homogenate pool. As both methods were shown to possess comparable sensitivities, we propose the C-BSE PMCA as a potential in-vitro replacement method, allowing the reduction and refinement of mouse bioassays for the detection of cattle derived classical BSE prions by reducing them to only specific analytical applications. <\/p>

Keywords<\/span>
Bovine spongiform encephalopathy, infectivity, PrPBSE<\/span>, Protein Misfolding Cyclic Amplification, replacement of animal experiments<\/p>

Zusammenfassung<\/span>
Die klassische Bovine Spongiforme Enzephalopathie (C-BSE) geh\u00f6rt zu den Transmissiblen Spongiformen Enzephalopathien (TSE), welche auch als Prion-Erkrankungen bezeichnet werden, da ihnen die Konversion des wirtseigenen zellul\u00e4ren Prion-Proteins PrPC<\/span> in seine pathologische Isoform PrPTSE<\/span> zugrunde liegt. BSE birgt ein zoonotisches Risiko, da die neue Variante der Creutzfeldt-Jakob-Krankheit beim Menschen durch BSE-Prionen hervorgerufen wird. Bislang kann C-BSE-Infektiosit\u00e4t nur mittels Bioassay, z. B. in Rinder-PrP-transgenen M\u00e4usen (z. B. Tgbov XV M\u00e4usen) nachgewiesen werden. Unl\u00e4ngst wurden auf der Keimbildungsaktivit\u00e4t der Prionen basierende hochsensitive In-vitro-Methoden, wie die Protein Misfolding Cyclic Amplification (PMCA), entwickelt. Diese sind besonders effektiv in der In-vitro-Konversion von Scrapie-Prionen, w\u00e4hrend eine \u00e4hnlich effiziente Amplifikation von bovinen C-BSE-Prionen bisher nicht zur Verf\u00fcgung stand. In der hier beschriebenen Studie wurde daher die analytische Sensitivit\u00e4t des transgenen Tgbov XV-Maus-Bioassays und unseres C-BSE-PMCA-Protokolls mittels Untersuchung einer seriellen Verd\u00fcnnungsreihe eines BSE-positivem Hirnstamm-Homogenat-Pools verglichen. Aufgrund der so nachgewiesenen vergleichbaren Sensitivit\u00e4ten beider Methoden schlagen wir die C-BSE-PMCA als potentielle In-vitro-Ersatzmethode vor, die die Reduktion und Verbesserung (Refinement) des transgenen Maus-Bioassays zum Nachweis von Rinder-C-BSE-Prionen erlaubt, da dieser auf spezielle analytische Anwendungen reduziert werden kann.<\/p>

Schl\u00fcsselw\u00f6rter<\/span>
Bovine Spongiforme Enzephalopathie, Infektiosit\u00e4t, PrPBSE<\/span>, Protein Misfolding Cyclic Amplification, Ersatzmethoden zum Tierversuch<\/p>","categories":["Open Access","Tier\u00e4rztliche Wochenschrift","Abostufe BMTW","Fachartikel","Abostufe frei"],"fromDate":"Jun 25, 2018 12:40:25 PM","oldUrls":["http:\/\/vetline.de\/exploring-pmca-as-a-potential-in-vitro-alternative-method-to-mouse-bioassays-for-the-highly-sensitive-detection-of-bse-prions\/150\/3130\/108497","http:\/\/vetline.de\/exploring-pmca-as-a-potential-in-vitro-alternative-method-to-mouse-bioassays-for-the-highly-sensitive-detection-of-bse-prions\/150\/3216\/108497"],"doiLanguage":"englisch","doiProductFormat":"online","doiPublisher":"Schl\u00fctersche Verlagsgesellschaft mbH & Co. KG","doiSerialWorkTitle":"Berl M\u00fcnch Tier\u00e4rztl Wochensch","doiDocumentUri":"https:\/\/vetline.de\/files\/smfiledata\/7\/2\/3\/5\/8\/4\/BMTW_OA_18021_BalkemaBuschmann.pdf","doiSource":"Berl M\u00fcnch Tier\u00e4rztl Wochensch","doiissn":"0005-9366","doiNr":"10.2376\/0005-9366-18021","doiFirstPage":".","doiLastPage":"..","doiTransmitted":true,"doiAuthor":"Ackermann I, Shawulu JC, Keller M, Fatola OI, Groschup MH, Balkema-Buschmann A","pdf":{"path":"http:\/\/data\/BMTW_OA_18021_BalkemaBuschmann.pdf","title":"BMTW_OA_18021_Balkema-Buschmann","description":"Exploring PMCA as a potential in-vitro alternative method to mouse bioassays for the highly sensitive detection of BSE prions"},"authors":[{"firstName":"I","middleName":"","lastName":"Ackermann"},{"firstName":"J","middleName":"C","lastName":"Shawulu"},{"firstName":"M","middleName":"","lastName":"Keller"},{"firstName":"O","middleName":"I","lastName":"Fatola"},{"firstName":"M","middleName":"H","lastName":"Groschup"},{"firstName":"A","middleName":"","lastName":"Balkema-Buschmann"}],"contentOptimised":"

Summary<\/strong>
Classical bovine spongiform encephalopathy (C-BSE) belongs to the transmissible spongiform encephalopathies (TSE), which are also designated prion diseases since they are caused by the conversion of the host-encoded cellular prion protein PrPC to its pathological isoform PrPTSE. BSE carries a zoonotic potential as BSE prions cause variant Creutzfeldt-Jakob disease in humans. To date, C-BSE infectivity can only be detected by bioassay, e.g. highly sensitive bovine PrP transgenic mice (e.g. Tgbov XV mice). Recently, highly sensitive in-vitro prion seeding activity assays, such as the Protein Misfolding Cyclic Amplification (PMCA), have been developed, which work particularly well for the template-assisted prion conversion of scrapie prions, while a similarly efficient bovine C-BSE-prion amplification remained unavailable. In the here described study, we have therefore compared the analytical sensitivities of the transgenic Tgbov XV mouse bioassay and our C-BSE PMCA protocol by analysing serial dilutions of a BSE-positive bovine brainstem homogenate pool. As both methods were shown to possess comparable sensitivities, we propose the C-BSE PMCA as a potential in-vitro replacement method, allowing the reduction and refinement of mouse bioassays for the detection of cattle derived classical BSE prions by reducing them to only specific analytical applications. <\/p>

Keywords:<\/strong>
Bovine spongiform encephalopathy, infectivity, PrPBSE, Protein Misfolding Cyclic Amplification, replacement of animal experiments<\/p>

Zusammenfassung<\/strong>
Die klassische Bovine Spongiforme Enzephalopathie (C-BSE) geh\u00f6rt zu den Transmissiblen Spongiformen Enzephalopathien (TSE), welche auch als Prion-Erkrankungen bezeichnet werden, da ihnen die Konversion des wirtseigenen zellul\u00e4ren Prion-Proteins PrPC in seine pathologische Isoform PrPTSE zugrunde liegt. BSE birgt ein zoonotisches Risiko, da die neue Variante der Creutzfeldt-Jakob-Krankheit beim Menschen durch BSE-Prionen hervorgerufen wird. Bislang kann C-BSE-Infektiosit\u00e4t nur mittels Bioassay, z. B. in Rinder-PrP-transgenen M\u00e4usen (z. B. Tgbov XV M\u00e4usen) nachgewiesen werden. Unl\u00e4ngst wurden auf der Keimbildungsaktivit\u00e4t der Prionen basierende hochsensitive In-vitro-Methoden, wie die Protein Misfolding Cyclic Amplification (PMCA), entwickelt. Diese sind besonders effektiv in der In-vitro-Konversion von Scrapie-Prionen, w\u00e4hrend eine \u00e4hnlich effiziente Amplifikation von bovinen C-BSE-Prionen bisher nicht zur Verf\u00fcgung stand. In der hier beschriebenen Studie wurde daher die analytische Sensitivit\u00e4t des transgenen Tgbov XV-Maus-Bioassays und unseres C-BSE-PMCA-Protokolls mittels Untersuchung einer seriellen Verd\u00fcnnungsreihe eines BSE-positivem Hirnstamm-Homogenat-Pools verglichen. Aufgrund der so nachgewiesenen vergleichbaren Sensitivit\u00e4ten beider Methoden schlagen wir die C-BSE-PMCA als potentielle In-vitro-Ersatzmethode vor, die die Reduktion und Verbesserung (Refinement) des transgenen Maus-Bioassays zum Nachweis von Rinder-C-BSE-Prionen erlaubt, da dieser auf spezielle analytische Anwendungen reduziert werden kann.<\/p>

Schl\u00fcsselw\u00f6rter:<\/strong>
Bovine Spongiforme Enzephalopathie, Infektiosit\u00e4t, PrPBSE, Protein Misfolding Cyclic Amplification, Ersatzmethoden zum Tierversuch<\/p>","primaryLanguage":"englisch","summary":"Classical bovine spongiform encephalopathy (C-BSE) belongs to the transmissible spongiform encephalopathies (TSE), which are also designated prion diseases since they are caused by the conversion of the host-encoded cellular prion protein PrPC to its pathological isoform PrPTSE. BSE carries a zoonotic potential as BSE prions cause variant Creutzfeldt-Jakob disease in humans. To date, C-BSE infectivity can only be detected by bioassay, e.g. highly sensitive bovine PrP transgenic mice (e.g. Tgbov XV mice). Recently, highly sensitive in-vitro prion seeding activity assays, such as the Protein Misfolding Cyclic Amplification (PMCA), have been developed, which work particularly well for the template-assisted prion conversion of scrapie prions, while a similarly efficient bovine C-BSE-prion amplification remained unavailable. In the here described study, we have therefore compared the analytical sensitivities of the transgenic Tgbov XV mouse bioassay and our C-BSE PMCA protocol by analysing serial dilutions of a BSE-positive bovine brainstem homogenate pool. As both methods were shown to possess comparable sensitivities, we propose the C-BSE PMCA as a potential in-vitro replacement method, allowing the reduction and refinement of mouse bioassays for the detection of cattle derived classical BSE prions by reducing them to only specific analytical applications. <\/p>

","keywords":["Bovine spongiform encephalopathy","infectivity","PrPBSE","Protein Misfolding Cyclic Amplification","replacement of animal experiments"],"zusammenfassung":"Die klassische Bovine Spongiforme Enzephalopathie (C-BSE) geh\u00f6rt zu den Transmissiblen Spongiformen Enzephalopathien (TSE), welche auch als Prion-Erkrankungen bezeichnet werden, da ihnen die Konversion des wirtseigenen zellul\u00e4ren Prion-Proteins PrPC in seine pathologische Isoform PrPTSE zugrunde liegt. BSE birgt ein zoonotisches Risiko, da die neue Variante der Creutzfeldt-Jakob-Krankheit beim Menschen durch BSE-Prionen hervorgerufen wird. Bislang kann C-BSE-Infektiosit\u00e4t nur mittels Bioassay, z. B. in Rinder-PrP-transgenen M\u00e4usen (z. B. Tgbov XV M\u00e4usen) nachgewiesen werden. Unl\u00e4ngst wurden auf der Keimbildungsaktivit\u00e4t der Prionen basierende hochsensitive In-vitro-Methoden, wie die Protein Misfolding Cyclic Amplification (PMCA), entwickelt. Diese sind besonders effektiv in der In-vitro-Konversion von Scrapie-Prionen, w\u00e4hrend eine \u00e4hnlich effiziente Amplifikation von bovinen C-BSE-Prionen bisher nicht zur Verf\u00fcgung stand. In der hier beschriebenen Studie wurde daher die analytische Sensitivit\u00e4t des transgenen Tgbov XV-Maus-Bioassays und unseres C-BSE-PMCA-Protokolls mittels Untersuchung einer seriellen Verd\u00fcnnungsreihe eines BSE-positivem Hirnstamm-Homogenat-Pools verglichen. Aufgrund der so nachgewiesenen vergleichbaren Sensitivit\u00e4ten beider Methoden schlagen wir die C-BSE-PMCA als potentielle In-vitro-Ersatzmethode vor, die die Reduktion und Verbesserung (Refinement) des transgenen Maus-Bioassays zum Nachweis von Rinder-C-BSE-Prionen erlaubt, da dieser auf spezielle analytische Anwendungen reduziert werden kann.<\/p>

","schluesselwoerter":["Bovine Spongiforme Enzephalopathie","Infektiosit\u00e4t","PrPBSE","Protein Misfolding Cyclic Amplification","Ersatzmethoden zum Tierversuch"],"translatedTitle":"Evaluierung der PMCA als In-vitro-Ersatzmethode zum Maus-Bioassay zur hochsensitiven Detektion von BSE-Prionen","abstractE":"Classical bovine spongiform encephalopathy (C-BSE) belongs to the transmissible spongiform encephalopathies (TSE), which are also designated prion diseases since they are caused by the conversion of the host-encoded cellular prion protein PrPC to its pathological isoform PrPTSE. BSE carries a zoonotic potential as BSE prions cause variant Creutzfeldt-Jakob disease in humans. To date, C-BSE infectivity can only be detected by bioassay, e.g. highly sensitive bovine PrP transgenic mice (e.g. Tgbov XV mice). Recently, highly sensitive in-vitro prion seeding activity assays, such as the Protein Misfolding Cyclic Amplification (PMCA), have been developed, which work particularly well for the template-assisted prion conversion of scrapie prions, while a similarly efficient bovine C-BSE-prion amplification remained unavailable. In the here described study, we have therefore compared the analytical sensitivities of the transgenic Tgbov XV mouse bioassay and our C-BSE PMCA protocol by analysing serial dilutions of a BSE-positive bovine brainstem homogenate pool. As both methods were shown to possess comparable sensitivities, we propose the C-BSE PMCA as a potential in-vitro replacement method, allowing the reduction and refinement of mouse bioassays for the detection of cattle derived classical BSE prions by reducing them to only specific analytical applications. ","date":{"year":2018,"date":"06\/2018","accepted":"2018-06-25"},"volume":131,"openAccess":true,"journal":"Berliner und M\u00fcnchener Tier\u00e4rztliche Wochenschrift","titleImageId":944,"pages":"","redirects":["exploring-pmca-as-a-potential-in-vitro-alternative-method-to-mouse-bioassays-for-the-highly-sensitive-detection-of-bse-prions\/150\/3130\/108497","exploring-pmca-as-a-potential-in-vitro-alternative-method-to-mouse-bioassays-for-the-highly-sensitive-detection-of-bse-prions\/150\/3216\/108497"],"tierartCategories":[],"artikelartCategories":["Open Access","Tier\u00e4rztliche Wochenschrift","Abostufe BMTW","Fachartikel","Abostufe frei"]} CY - Hannover DA - 06/2018 DO - 10.2376/0005-9366-18021 LA - English N2 - Classical bovine spongiform encephalopathy (C-BSE) belongs to the transmissible spongiform encephalopathies (TSE), which are also designated prion diseases since they are caused by the conversion of the host-encoded cellular prion protein PrPC to its pathological isoform PrPTSE. BSE carries a zoonotic potential as BSE prions cause variant Creutzfeldt-Jakob disease in humans. To date, C-BSE infectivity can only be detected by bioassay, e.g. highly sensitive bovine PrP transgenic mice (e.g. Tgbov XV mice). Recently, highly sensitive in-vitro prion seeding activity assays, such as the Protein Misfolding Cyclic Amplification (PMCA), have been developed, which work particularly well for the template-assisted prion conversion of scrapie prions, while a similarly efficient bovine C-BSE-prion amplification remained unavailable. In the here described study, we have therefore compared the analytical sensitivities of the transgenic Tgbov XV mouse bioassay and our C-BSE PMCA protocol by analysing serial dilutions of a BSE-positive bovine brainstem homogenate pool. As both methods were shown to possess comparable sensitivities, we propose the C-BSE PMCA as a potential in-vitro replacement method, allowing the reduction and refinement of mouse bioassays for the detection of cattle derived classical BSE prions by reducing them to only specific analytical applications. PB - Schlütersche Verlagsgesellschaft mbH & Co. KG PP - Hannover PY - 2018 T1 - Exploring PMCA as a potential in-vitro alternative method to mouse bioassays for the highly sensitive detection of BSE prions T2 - Berliner und Münchener Tierärztliche Wochenschrift TI - Exploring PMCA as a potential in-vitro alternative method to mouse bioassays for the highly sensitive detection of BSE prions TT - Evaluierung der PMCA als In-vitro-Ersatzmethode zum Maus-Bioassay zur hochsensitiven Detektion von BSE-Prionen VL - 131 SN - 0005-9366 ER -