NR ATAT
AU Gofflot,S.; Deprez,M.; El Moualij,B.; Osman,A.; Thonnart,J.F.; Hougrand,O.; Heinen,E.; Zorzi,W.
TI Immunoquantitative PCR for prion protein detection in sporadic Creutzfeldt-Jakob disease
QU Clinical Chemistry 2005 Sep; 51(9): 1605-11
PT journal article
AB BACKGROUND: 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). METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
IN Die Autoren binden eine in Mensch und Tier nicht vorkommende Hybrid-DNA an die Antikörper, welche im ELISA das bereits über Antikörper an die Plastikoberfläche gebundene Prionprotein markieren. Nachdem nicht spezifisch an Prionprotein gebundene Antikörper abgewaschen wurden, vermehrt man durch PCR die Marker-DNA an den gebundenen Antikörpern. Durch diesen Trick konnten die Autoren die Empfindlichkeit des ELISA um mindestens eine Größenordnung steigern.
MH Adolescent; Aged; Aged, 80 and over; Blotting, Western; Brain Chemistry; Comparative Study; Creutzfeldt-Jakob Syndrome/*diagnosis; Endopeptidase K/metabolism; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunohistochemistry; Male; Middle Aged; Polymerase Chain Reaction; Prions/*analysis/metabolism; Research Support, Non-U.S. Gov't; Sensitivity and Specificity
AD Centre de Recherche sur les Proteines Prions (CRPP) - Service d'Histologie Humaine, Universite de Liege-CHU, Liege, Belgium.
SP englisch
PO USA