Immunoquantitative PCR for prion protein detection in sporadic Creutzfeldt-Jakob disease.
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.
Centre de Recherche sur les Protéines Prions (CRPP) - Service d'Histologie Humaine, Université de Liège-CHU, Liège, Belgium.
Stéphanie
S
Gofflot
Manuel
M
Deprez
Benaïssa
B
el Moualij
Awad
A
Osman
Jean-François
JF
Thonnart
Olivier
O
Hougrand
Ernst
E
Heinen
Willy
W
Zorzi
D000293:Adolescent; D000368:Aged; D000369:Aged, 80 and over; D015153:Blotting, Western; D001923:Brain Chemistry; D007562:Creutzfeldt-Jakob Syndrome / Q000175:diagnosis*; D019286:Endopeptidase K / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D011328:Prions / Q000032:analysis* / Q000378:metabolism; D012680:Sensitivity and Specificity
D011328:Prions; D019286:Endopeptidase K