Christina D Orrú1, Matilde Bongianni, Giovanni Tonoli, Sergio Ferrari, Andrew G Hughson, Bradley R Groveman, Michele Fiorini, Maurizio Pocchiari, Salvatore Monaco, Byron Caughey, Gianluigi Zanusso. 1. From the Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health, Hamilton, MT (C.D.O., M.B., A.G.H., B.R.G., B.C.); and the Department of Biomedical Sciences, University of Cagliari, Cagliari (C.D.O.), the Department of Neurologic and Movement Sciences, University of Verona, Verona (M.B., S.F., M.F., S.M., G.Z.), Clinica Otorinolaringoiatrica, Policlinico G.B. Rossi, Azienda Ospedaliera Universitaria Integrata, Verona (G.T.), and the Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, Rome (M.P.) - all in Italy.
Abstract
BACKGROUND: Definite diagnosis of sporadic Creutzfeldt-Jakob disease in living patients remains a challenge. A test that detects the specific marker for Creutzfeldt-Jakob disease, the prion protein (PrP(CJD)), by means of real-time quaking-induced conversion (RT-QuIC) testing of cerebrospinal fluid has a sensitivity of 80 to 90% for the diagnosis of sporadic Creutzfeldt-Jakob disease. We have assessed the accuracy of RT-QuIC analysis of nasal brushings from olfactory epithelium in diagnosing sporadic Creutzfeldt-Jakob disease in living patients. METHODS: We collected olfactory epithelium brushings and cerebrospinal fluid samples from patients with and patients without sporadic Creutzfeldt-Jakob disease and tested them using RT-QuIC, an ultrasensitive, multiwell plate-based fluorescence assay involving PrP(CJD)-seeded polymerization of recombinant PrP into amyloid fibrils. RESULTS: The RT-QuIC assays seeded with nasal brushings were positive in 30 of 31 patients with Creutzfeldt-Jakob disease (15 of 15 with definite sporadic Creutzfeldt-Jakob disease, 13 of 14 with probable sporadic Creutzfeldt-Jakob disease, and 2 of 2 with inherited Creutzfeldt-Jakob disease) but were negative in 43 of 43 patients without Creutzfeldt-Jakob disease, indicating a sensitivity of 97% (95% confidence interval [CI], 82 to 100) and specificity of 100% (95% CI, 90 to 100) for the detection of Creutzfeldt-Jakob disease. By comparison, testing of cerebrospinal fluid samples from the same group of patients had a sensitivity of 77% (95% CI, 57 to 89) and a specificity of 100% (95% CI, 90 to 100). Nasal brushings elicited stronger and faster RT-QuIC responses than cerebrospinal fluid (P<0.001 for the between-group comparison of strength of response). Individual brushings contained approximately 10(5) to 10(7) prion seeds, at concentrations several logs10 greater than in cerebrospinal fluid. CONCLUSIONS: In this preliminary study, RT-QuIC testing of olfactory epithelium samples obtained from nasal brushings was accurate in diagnosing Creutzfeldt-Jakob disease and indicated substantial prion seeding activity lining the nasal vault. (Funded by the Intramural Research Program of the National Institute of Allergy and Infectious Diseases and others.).
BACKGROUND: Definite diagnosis of sporadic Creutzfeldt-Jakob disease in living patients remains a challenge. A test that detects the specific marker for Creutzfeldt-Jakob disease, the prion protein (PrP(CJD)), by means of real-time quaking-induced conversion (RT-QuIC) testing of cerebrospinal fluid has a sensitivity of 80 to 90% for the diagnosis of sporadic Creutzfeldt-Jakob disease. We have assessed the accuracy of RT-QuIC analysis of nasal brushings from olfactory epithelium in diagnosing sporadic Creutzfeldt-Jakob disease in living patients. METHODS: We collected olfactory epithelium brushings and cerebrospinal fluid samples from patients with and patients without sporadic Creutzfeldt-Jakob disease and tested them using RT-QuIC, an ultrasensitive, multiwell plate-based fluorescence assay involving PrP(CJD)-seeded polymerization of recombinant PrP into amyloid fibrils. RESULTS: The RT-QuIC assays seeded with nasal brushings were positive in 30 of 31 patients with Creutzfeldt-Jakob disease (15 of 15 with definite sporadic Creutzfeldt-Jakob disease, 13 of 14 with probable sporadic Creutzfeldt-Jakob disease, and 2 of 2 with inherited Creutzfeldt-Jakob disease) but were negative in 43 of 43 patients without Creutzfeldt-Jakob disease, indicating a sensitivity of 97% (95% confidence interval [CI], 82 to 100) and specificity of 100% (95% CI, 90 to 100) for the detection of Creutzfeldt-Jakob disease. By comparison, testing of cerebrospinal fluid samples from the same group of patients had a sensitivity of 77% (95% CI, 57 to 89) and a specificity of 100% (95% CI, 90 to 100). Nasal brushings elicited stronger and faster RT-QuIC responses than cerebrospinal fluid (P<0.001 for the between-group comparison of strength of response). Individual brushings contained approximately 10(5) to 10(7) prion seeds, at concentrations several logs10 greater than in cerebrospinal fluid. CONCLUSIONS: In this preliminary study, RT-QuIC testing of olfactory epithelium samples obtained from nasal brushings was accurate in diagnosing Creutzfeldt-Jakob disease and indicated substantial prion seeding activity lining the nasal vault. (Funded by the Intramural Research Program of the National Institute of Allergy and Infectious Diseases and others.).
Authors: Richard A Bessen; Jason M Wilham; Diana Lowe; Christopher P Watschke; Harold Shearin; Scott Martinka; Byron Caughey; James A Wiley Journal: J Virol Date: 2011-11-30 Impact factor: 5.103
Authors: Jason M Wilham; Christina D Orrú; Richard A Bessen; Ryuichiro Atarashi; Kazunori Sano; Brent Race; Kimberly D Meade-White; Lara M Taubner; Andrew Timmes; Byron Caughey Journal: PLoS Pathog Date: 2010-12-02 Impact factor: 6.823
Authors: Piero Parchi; Laura de Boni; Daniela Saverioni; Mark L Cohen; Isidro Ferrer; Pierluigi Gambetti; Ellen Gelpi; Giorgio Giaccone; Jean-Jacques Hauw; Romana Höftberger; James W Ironside; Casper Jansen; Gabor G Kovacs; Annemieke Rozemuller; Danielle Seilhean; Fabrizio Tagliavini; Armin Giese; Hans A Kretzschmar Journal: Acta Neuropathol Date: 2012-06-30 Impact factor: 17.088
Authors: Paul Brown; Jean-Philippe Brandel; Takeshi Sato; Yosikazu Nakamura; Jan MacKenzie; Robert G Will; Anna Ladogana; Maurizio Pocchiari; Ellen W Leschek; Lawrence B Schonberger Journal: Emerg Infect Dis Date: 2012-06 Impact factor: 6.883
Authors: Lynne I McGuire; Alexander H Peden; Christina D Orrú; Jason M Wilham; Nigel E Appleford; Gary Mallinson; Mary Andrews; Mark W Head; Byron Caughey; Robert G Will; Richard S G Knight; Alison J E Green Journal: Ann Neurol Date: 2012-08 Impact factor: 10.422
Authors: Nicholas J Haley; Chris Siepker; W David Walter; Bruce V Thomsen; Justin J Greenlee; Aaron D Lehmkuhl; Jürgen A Richt Journal: J Clin Microbiol Date: 2016-02-10 Impact factor: 5.948
Authors: Eri Saijo; Michael A Metrick; Shunsuke Koga; Piero Parchi; Irene Litvan; Salvatore Spina; Adam Boxer; Julio C Rojas; Douglas Galasko; Allison Kraus; Marcello Rossi; Kathy Newell; Gianluigi Zanusso; Lea T Grinberg; William W Seeley; Bernardino Ghetti; Dennis W Dickson; Byron Caughey Journal: Acta Neuropathol Date: 2019-10-16 Impact factor: 17.088
Authors: Matthias Schmitz; Maria Cramm; Franc Llorens; Dominik Müller-Cramm; Steven Collins; Ryuichiro Atarashi; Katsuya Satoh; Christina D Orrù; Bradley R Groveman; Saima Zafar; Walter J Schulz-Schaeffer; Byron Caughey; Inga Zerr Journal: Nat Protoc Date: 2016-10-13 Impact factor: 13.491
Authors: Rohana P Dassanayake; Christina D Orrú; Andrew G Hughson; Byron Caughey; Telmo Graça; Dongyue Zhuang; Sally A Madsen-Bouterse; Donald P Knowles; David A Schneider Journal: J Gen Virol Date: 2015-12-10 Impact factor: 3.891
Authors: Kristen A Davenport; Brittany A Mosher; Brian M Brost; Davin M Henderson; Nathaniel D Denkers; Amy V Nalls; Erin McNulty; Candace K Mathiason; Edward A Hoover Journal: J Clin Microbiol Date: 2017-12-26 Impact factor: 5.948