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Estimating prion concentration in fluids and tissues by quantitative PMCA

Nature Methods volume 7, pages 519–520 (2010)Cite this article

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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.

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Figure 1: Relationship between PrPSc concentration and PMCA rounds required for detection.
Figure 2: Detection of PrPSc in spleen of scrapie-affected hamsters.

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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.

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Authors and Affiliations

  1. Department of Neurology, Mitchell Center for Alzheimer's Disease and Related Brain Disorders, University of Texas Houston Medical School, Houston, Texas, USA

    Baian Chen, Rodrigo Morales, Marcelo A Barria & Claudio Soto

  2. National Animal Transmissible Spongiform Encephalopathies (TSE) Lab, College of Veterinary Medicine, China Agricultural University,, Beijing, China

    Baian Chen

Authors
  1. Baian Chen
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  2. Rodrigo Morales
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  3. Marcelo A Barria
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  4. Claudio Soto
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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

Correspondence to Claudio Soto.

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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.

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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). https://doi.org/10.1038/nmeth.1465

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