Abstract
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.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Aguzzi, A. & Calella, A.M. Physiol. Rev. 89, 1105–1152 (2009).
Soto, C. Nat. Rev. Microbiol. 2, 809–819 (2004).
Brown, P. Neurology 70, 713–722 (2008).
Wadsworth, J.D. et al. Lancet 358, 171–180 (2001).
Saborio, G.P., Permanne, B. & Soto, C. Nature 411, 810–813 (2001).
Saa, P., Castilla, J. & Soto, C. J. Biol. Chem. 281, 35245–35252 (2006).
Castilla, J., Saa, P. & Soto, C. Nat. Med. 11, 982–985 (2005).
Saa, P., Castilla, J. & Soto, C. Science 313, 92–94 (2006).
Thorne, L. & Terry, L.A. J. Gen. Virol. 89, 3177–3184 (2008).
Murayama, Y. et al. J. Gen. Virol. 88, 2890–2898 (2007).
Gonzalez-Romero, D., Barria, M.A., Leon, P., Morales, R. & Soto, C. FEBS Lett. 582, 3161–3166 (2008).
Morales, R. et al. Biochem. Biophys. Res. Commun. 377, 373–378 (2008).
Castilla, J. et al. Methods Enzymol. 412, 3–21 (2006).
Castilla, J., Saá, P., Hetz, C. & Soto, C. Cell 121, 195–206 (2005).
Saa, P., Castilla, J. & Soto, C. Methods Mol. Biol. 299, 53–65 (2005).
Acknowledgements
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.
Author information
Authors and Affiliations
Contributions
B.C. performed most of the experiments, analyzed the data and prepared the figures; R.M. helped with experimental design and data analysis; M.A.B. performed the PMCA experiments in blood; and C.S. developed the hypothesis, designed and analyzed all data, wrote the manuscript and supervised the project.
Corresponding author
Ethics declarations
Competing interests
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.
Rights and permissions
About this article
Cite this article
Chen, B., Morales, R., Barria, M. et al. Estimating prion concentration in fluids and tissues by quantitative PMCA. Nat Methods 7, 519–520 (2010). https://doi.org/10.1038/nmeth.1465
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nmeth.1465
This article is cited by
-
Detection of prions from spiked and free-ranging carnivore feces
Scientific Reports (2024)
-
Ticks harbor and excrete chronic wasting disease prions
Scientific Reports (2023)
-
PMCA screening of retropharyngeal lymph nodes in white-tailed deer and comparisons with ELISA and IHC
Scientific Reports (2023)
-
PMCA for ultrasensitive detection of prions and to study disease biology
Cell and Tissue Research (2023)
-
Detection of CWD prions in naturally infected white-tailed deer fetuses and gestational tissues by PMCA
Scientific Reports (2021)