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Clin Chem ; 51(9): 1605-11, 2005 Sep.
Article in English | MEDLINE | ID: mdl-16002456

ABSTRACT

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.


Subject(s)
Creutzfeldt-Jakob Syndrome/diagnosis , Prions/analysis , Adolescent , Aged , Aged, 80 and over , Blotting, Western , Brain Chemistry , Endopeptidase K/metabolism , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunohistochemistry , Male , Middle Aged , Polymerase Chain Reaction , Prions/metabolism , Sensitivity and Specificity
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