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Fast and ultrasensitive method for quantitating prion infectivity titre.
Natallia Makarava1, Regina Savtchenko, Irina Alexeeva
1Center for Biomedical Engineering and Technology, University of Maryland, 725 W. Lombard Street, Baltimore 21201, USA.
Nature Communications
|March 15, 2012
Summary
The protein misfolding cyclic amplification with beads (PMCAb) technique precisely estimates prion protein titres in days, significantly faster than traditional bioassays. This method offers a more efficient and cost-effective approach for prion disease research.
Area of Science:
- Neuroscience
- Biochemistry
- Molecular Biology
Background:
- Prion infectivity titre determination traditionally relies on end-point dilution bioassays.
- These bioassays are time-consuming, often requiring up to two years for results.
Purpose of the Study:
- To evaluate the efficacy of the protein misfolding cyclic amplification with beads (PMCAb) technique for prion titre estimation.
- To compare the precision and speed of PMCAb with conventional bioassays.
Main Methods:
- Utilized the PMCAb technique to estimate prion titres in hamster brain tissue.
- Compared PMCAb-derived median reactive doses (PMCAb(50)) with bioassay-derived median infectious doses (ID(50)).
Main Results:
- PMCAb determined prion titres with higher precision and in 3-6 days, compared to the 2-year bioassay.
- For hamster strains 263K and SSLOW, PMCAb(50) values were 10(12.8) and 10(12.2) per gram, respectively.
- PMCAb(50) values were significantly higher (160- to 4,000-fold) than corresponding ID(50) values.
Conclusions:
- PMCAb is a precise, rapid, and potentially cost-effective method for estimating infection-specific prion protein titres.
- Discrepancies between PMCAb and bioassay titres may indicate an excess of non-infectious seeds or differential clearance rates.
