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Detection of misfolded prion protein in blood with conformationally sensitive peptides
Author(s) -
Pan Tao,
Sethi Jasmeet,
Nelsen Craig,
Rudolph Alan,
Cervenakova Larisa,
Brown Paul,
Orser Cindy S.
Publication year - 2007
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/j.1537-2995.2007.01284.x
Subject(s) - transmissible spongiform encephalopathy , prion protein , microbiology and biotechnology , virology , peptide , scrapie , chemistry , biology , medicine , pathology , biochemistry , disease
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 β‐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‐µL 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.