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Prions and Prion Diseases: Quaking-induced conversion (QuIC) assays for the detection and diagnosis of TSE prion diseases.
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Ultrasensitive detection and quantitation of prion seeding activity by RT-QuIC and eQuIC assays.
No full textUltrasensitive detection of prion seeding activity by RT-QuIC and eQuIC assays for early diagnosis of prion diseases. 15° Convegno degli Istituti Zooprofilattici Sperimentali sulle Encefalopatie Spongiformi Animali, Sala Conferenze Istituto Zooprofilattico Sperimentale del Piemonte Liguria e Valle d’Aosta, Torino (Italia), 4-5 Ottobre 2012.
No full textNew generation QuIC assays for prion seeding activity
No full textThe ability of abnormal TSE-associated forms of PrP to seed the formation of amyloid fibrils from recombinant PrPSen has served as the basis for several relatively rapid and highly sensitive tests for prion diseases. These tests include rPrP-PMCA (rPMCA), standard quaking-induced conversion (S-QuIC), amyloid seeding assay (ASA), real-time QuIC (RT-QuIC) and enhanced QuIC (eQuIC). Here, we summarize recent improvements in the RT-QuIC-based assays that enhance the practicality, sensitivity and quantitative attributes of assays QuIC and promote the detection of prion seeding activity in dilute, inhibitor-laden fluids such as blood plasma
Ultrasensitive detection and quantitation of prion seeding activity by RT-QuIC and eQuIC assays. XII Congresso Federazione Italiana Scienze della Vita (FISV). Universita` La Sapienza Roma (Italia), 24-27 Settembre 2012.
No full textPrion disease blood test using immunoprecipitation and improved quaking-induced conversion
No full textA 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 10 14-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 (eQuIC) 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
Ultrasensitive detection and quantitation of prion seeding activity by RT-QuIC and eQuIC assays. 11o Congresso Nazionale della Societa` Italiana di Virologia, Palazzo del Capitano del Popolo, Orvieto (TR, Italia) 17-19 Settembre 2012.
No full textHuman variant Creutzfeldt-Jakob disease and sheep scrapie PrP(res) detection using seeded conversion of recombinant prion protein
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