PrPTSE detection


RT-QuIC and mass spectrometry approaches to prion strain detection and discrimination

2014: 18 months / Grant: 60 000 euros

Project: The goal of this project was to determine whether it was possible to discriminate prion strains by RT-QuIC or RT-QuIC combined with mass spectrometric analysis, which had been shown to differenciate animal prion strains. If it works, this could allow the distinction between type I and II CJD, or BSE and BASE, which would be relevant for surveillance in animals and diagnosis in humans.

Main results and related published data: By playing with different substrate sources of recombinant PrP (rPrP: Syrian golden hamster rPrP23-231 or rPrP90-231, human rPrP23-231), and chimeric hamster-sheep rPrP(23-137hamster+141-234ovine), it has been possible to discriminate the L-BSE agent from the classical C-BSE agent in RT-QuIC assay. The amplification of PrPTSE from CBSE could be improved and obtained only with a particular substrate (hamster-sheep rPrP), whereas the amplification of PrPTSE from L-BSE could be obtained using multiple rPrP (hamster-sheep rPrP, hamster rPrP 23-231 and 90-231, human 23-231). In addition, the limit of dilution with the hamstersheep rPrP substrate is higher with L-BSE compared to C-BSE. Results reported recently completed this study, since they showed that it was possible to discriminate the atypical H-BSE using multiple rPrP subtrates (bank vole, ovine, chimeric hamster bovine… rPrP). Importantly bank vole rPrP also appeared to be a virtually universalsubstrate for PrPTSE amplification using the RT-QuIC method for many prion sources (vCJD, GSS, sCJD, several animal TSE and experimental models….).