Prions, the proteinaceous infectious agent responsible for prion diseases, can be detected with high sensitivity by protein misfolding cyclic amplification (PMCA) technology. Here we describe a quantitative PMCA procedure to calculate the concentration of very low levels of prions in biological samples. Using this procedure, we determined the quantities of misfolded prion protein (PrPSc) in brain, spleen, blood and urine of scrapie-affected hamsters.
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We thank D. Gonzalez-Romero (University of Texas Houston Medical School) for participating in the PMCA experiments with urine, R. Diaz-Espinoza (University of Texas Houston Medical School) for providing recombinant PrP and D. Zhao (China Agricultural University) for suggestions and providing support to B.C. This research was supported in part by US National Institutes of Health grants R01NS049173 and P01AI077774 to C.S.
C.S. is inventor of the PMCA technology and is founder, vice-president and chief scientific officer of Amprion, Inc, a biotechnology company focused on developing technologies to detect misfolded proteins associated with neurodegenerative diseases.
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Chen, B., Morales, R., Barria, M. et al. Estimating prion concentration in fluids and tissues by quantitative PMCA. Nat Methods 7, 519–520 (2010) doi:10.1038/nmeth.1465
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