NR AXGS
AU Pan,T.; Sethi,J.; Nelsen,C.; Rudolph,A.; Cervenakova,L.; Brown,P.; Orser,C.S.
TI Detection of misfolded prion protein in blood with conformationally sensitive peptides
QU Transfusion 2007 Aug; 47(8): 1418-25
PT journal article; research support, n.i.h., extramural; research support, u.s. gov't, non-p.h.s.
AB BACKGROUND: The long-standing goal of a preclinical diagnostic test for transmissible spongiform encephalopathy (TSE) has recently become urgent because of the discovery that humans with variant Creutzfeldt-Jakob disease can transmit disease via blood transfusions. STUDY DESIGN AND METHODS: The misfolded protein diagnostic (MPD) assay employs a pyrene-labeled palindromic sequence of prion peptides that undergoes a cascade of coil to beta-sheet conversion in the presence of the misfolded prion protein (PrP(TSE)). The ability of the assay to detect PrP(TSE) in brain, serum, and plasma was tested. The basic protocol involved a several-hour incubation of 200-microL sample volumes with the peptide reagent in 96-well plates, after which fluorescence was monitored by a fluorescence plate reader with an excitation wavelength of 350 nm and emission scanning wavelength range of 365 to 600 nm. RESULTS: Target specificity for PrP(TSE) was documented by correlation of assay signal with Western blot signals in brain tissue from TSE-infected, normal, and knockout mice and negative assay signals by use of reagents with different peptide sequences. When applied to plasma or serum, the assay discriminated between samples from a variety of experimental and natural TSE infections compared to uninfected controls, with a sensitivity threshold of approximately 1 infectious dose per mL in pooled plasma from TSE-infected mice. CONCLUSIONS: The MPD assay is a sensitive and specific test for the detection of PrP(TSE) that may be useful in both preclinical and clinical diagnosis of TSE diseases of animals and humans.
MH Amino Acid Sequence; Animals; Creutzfeldt-Jakob Syndrome; Disease Models, Animal; Fluorescence; Humans; Mice; Molecular Sequence Data; Prions/*blood/*chemistry; Protein Conformation; *Protein Folding; Saimiri; Sensitivity and Specificity; Sheep
AD Adlyfe Inc., 9430 Key West Avenue, Rockville, MD 20850, USA.
SP englisch
PO USA