Immunoquantitative PCR for Prion Protein Detection in Sporadic Creutzfeldt-Jakob Disease

• Published in: Clinical chemistry (Baltimore, Md.) Vol. 51; no. 9; pp. 1605 - 1611
• Main Authors: Gofflot, Stephanie, Deprez, Manuel, el Moualij, Benaissa, Osman, Awad, Thonnart, Jean-Francois, Hougrand, Olivier, Heinen, Ernst, Zorzi, Willy
• Format: Journal Article Web Resource
• Language: English
• Published: Washington, DC Am Assoc Clin Chem 01.09.2005; American Association for Clinical Chemistry; Oxford University Press
• Subjects: Adolescent; Aged; Aged, 80 and over; Analytical, structural and metabolic biochemistry; Anatomie (cytologie, histologie, embryologie...) & physiologie; Anatomy (cytology, histology, embryology...) & physiology; Biochemistry, biophysics & molecular biology; Biochimie, biophysique & biologie moléculaire; Biological and medical sciences; Blotting, Western; Brain Chemistry; Creutzfeldt-Jakob disease; Creutzfeldt-Jakob Syndrome - diagnosis; Endopeptidase K - metabolism; Enzyme-Linked Immunosorbent Assay; Female; Fundamental and applied biological sciences. Psychology; Human health sciences; Humans; Immunohistochemistry; Investigative techniques, diagnostic techniques (general aspects); Laboratory medicine & medical technology; Life sciences; Male; Medical sciences; Middle Aged; Médecine de laboratoire & technologie médicale; Polymerase Chain Reaction; Prions - analysis; Prions - metabolism; Prions/analysis/metabolism; Sciences de la santé humaine; Sciences du vivant; Sensitivity and Specificity; Sterilization; Prion disease; Nervous system diseases; Sporadic; Biochemistry; Protein; Cerebral disorder; Infection; Polymerase chain reaction; Central nervous system disease; Clinical biology; Degenerative disease; Prion; Diagnosis; Molecular biology; Creutzfeldt Jakob disease
• ISSN: 0009-9147; 1530-8561; 1530-8561
• DOI: 10.1373/clinchem.2005.050120

The most common human prion disorder is Creutzfeldt-Jakob disease (CJD); it includes sporadic, familial, iatrogenic, and variant subtypes. Diagnostic tests aim at detection with the highest specificity of very small deposits of abnormal prion protein (PrP). We used immunoquantitative PCR (iqPCR) to detect proteinase K-resistant PrP (PrPRes) in tissue from the middle frontal gyrus of 7 patients with sporadic CJD and 7 non-CJD cases. We compared iqPCR with routine optimized ELISA, Western blotting, and immunohistochemical analyses. The 4 methods showed similar 100% sensitivity and specificity for the diagnosis of CJD. Along with high specificity, however, iqPCR had a threshold for PrP(Res) detection at least 10-fold lower than that of the classic ELISA. iqPCR is a new method for PrPRes detection that combines 100% specificity with a detection threshold at least 10-fold lower than classic techniques. This method may improve the detection of minute PrPRes deposits in tissues and body fluids and thus be useful for diagnostic and sterilization applications.