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Ultrasensitive human prion detection in cerebrospinal fluid by real-time quaking-induced conversion

Abstract

The development of technologies for the in vitro amplification of abnormal conformations of prion protein (PrPSc) has generated the potential for sensitive detection of prions. Here we developed a new PrPSc amplification assay, called real-time quaking-induced conversion (RT-QUIC), which allows the detection of ≥1 fg of PrPSc in diluted Creutzfeldt-Jakob disease (CJD) brain homogenate. Moreover, we assessed the technique first in a series of Japanese subjects and then in a blind study of 30 cerebrospinal fluid specimens from Australia, which achieved greater than 80% sensitivity and 100% specificity. These findings indicate the promising enhanced diagnostic capacity of RT-QUIC in the antemortem evaluation of suspected CJD.

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Figure 1: QUIC reactions induce PrPSc-dependent rHuPrP-res formation under GdnHCl-free conditions.

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References

  1. Prusiner, S.B. Proc. Natl. Acad. Sci. USA 95, 13363–13383 (1998).

    Article  CAS  Google Scholar 

  2. Aguzzi, A. & Calella, A.M. Physiol. Rev. 89, 1105–1152 (2009).

    Article  CAS  Google Scholar 

  3. Lasmezas, C.I. et al. Science 275, 402–405 (1997).

    Article  CAS  Google Scholar 

  4. Gambetti, P. et al. Ann. Neurol. 63, 697–708 (2008).

    Article  CAS  Google Scholar 

  5. Saborio, G.P., Permanne, B. & Soto, C. Nature 411, 810–813 (2001).

    Article  CAS  Google Scholar 

  6. Castilla, J. et al. Methods Enzymol. 412, 3–21 (2006).

    Article  CAS  Google Scholar 

  7. Atarashi, R. et al. Nat. Methods 4, 645–650 (2007).

    Article  CAS  Google Scholar 

  8. Colby, D.W. et al. Proc. Natl. Acad. Sci. USA 104, 20914–20919 (2007).

    Article  CAS  Google Scholar 

  9. Atarashi, R. et al. Nat. Methods 5, 211–212 (2008).

    Article  CAS  Google Scholar 

  10. Orru, C.D. et al. Protein Eng. Des. Sel. 22, 515–521 (2009).

    Article  CAS  Google Scholar 

  11. Kocisko, D.A. et al. Nature 370, 471–474 (1994).

    Article  CAS  Google Scholar 

  12. Toth, S.I., Smith, L.A. & Ahmed, S.A. J. Pharm. Sci. 98, 3302–3311 (2009).

    Article  CAS  Google Scholar 

  13. Collins, S.R., Douglass, A., Vale, R.D. & Weissman, J.S. PLoS Biol. 2, e321 (2004).

    Article  Google Scholar 

  14. Lansbury, P.T. Jr. & Caughey, B. Chem. Biol. 2, 1–5 (1995).

    Article  CAS  Google Scholar 

  15. Hsich, G., Kenney, K., Gibbs, C.J., Lee, K.H. & Harrington, M.G. N. Engl. J. Med. 335, 924–930 (1996).

    Article  CAS  Google Scholar 

  16. Zerr, I. et al. Ann. Neurol. 43, 32–40 (1998).

    Article  CAS  Google Scholar 

  17. Parchi, P. et al. Ann. Neurol. 46, 224–233 (1999).

    Article  CAS  Google Scholar 

  18. Satoh, K. et al. Lab. Invest. 90, 1637–1644 (2010).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank the members of the CJD Surveillance Committee in Japan for their support of this work, K. Yamaguchi and K. Kuwata for helping with the circular dichroism analysis of rPrP-sen, M. Tsujihata (Nagasaki Kita Hospital) for providing CSF samples and information about subjects and A. Yamakawa and A. Matsuo for technical assistance. The work was supported by the Global Centers of Excellence Program (F12); a grant-in-aid for science research (B; grant no. 20390287) from the Ministry of Education, Culture, Sports, Science and Technology of Japan; a grant for bovine spongiform encephalopathy research and a grant-in-aid of the Research Committee of Prion disease and Slow Virus Infection, from the Ministry of Health, Labor and Welfare of Japan.

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Authors

Contributions

R.A. designed the project, performed experiments and wrote the manuscript. K. Satoh, K. Sano, T.F., N.Y., D.I., T.M., T.N. and H.Y. performed experiments. K. Satoh, S.S., M.Y., H.M., T.K., G.K., A.M. and S.J.C. contributed to the collection of human specimens and provided information about subjects. N.N. supervised the project. K. Satoh, K. Sano, A.M., S.J.C. and N.N. helped with the editing of the manuscript.

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Correspondence to Ryuichiro Atarashi.

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The authors declare no competing financial interests.

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Supplementary Figures 1–3 and Supplementary Tables 1–4 and Supplementary Methods (PDF 913 kb)

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Atarashi, R., Satoh, K., Sano, K. et al. Ultrasensitive human prion detection in cerebrospinal fluid by real-time quaking-induced conversion. Nat Med 17, 175–178 (2011). https://doi.org/10.1038/nm.2294

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