NR AWHF
AU Huelamo,T.; Parra,B.; Castilla,J.; Nevado,M.J.; Anadon,E.; Soto,C.; Gomez-Tejedor,C.; Mayoral,T.
TI Highly increasing of sensitivity in scrapie detection by rapid tests using PMCA
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions DIA-27
PT Konferenz-Poster
AB PMCA (Protein Misfolding Cyclic Amplification) is a recently described technology for in vitro cycling amplification and aggregation of PrPres. In a similar way to DNA amplification by PCR, PMCA increases the presence of PrPres in a sample with initial minute quantity of PrPsc. Previous studies have shown evolution of PMCA from a manual to an automated method that improves reproducibility and efficiency. Also infectivity and PrPsc detection in blood has been demonstrated in scrapie-afflicted hamsters. In this work we will show how automated Scrapie PMCA can be linked to UE approved rapid tests to highly increase the sensitivity of them. Also PMCA can be used for PrPsc amplification obtained from nervous tissue and others as spleen. In vitro amplified PrPres, show similar characteristics to Scrapie PrPsc and can be detected by same antibodies that recognizes different epitopes of PrPsc protein and tests designed for specific capture of PrPsc show same behaviour using PrPres from PMCA as PrPsc from scrapie affected sheep tissues. Results will show that three runs (120 cycles) of automated Scrapie PMCA leads to a 15000 fold increase in sensitivity compared to standard diagnosis methods in nervous tissue.
AD T. Huelamo, B. Parra, M.J. Nevado, E. Anadón, C. Gómez-Tejedor, T. Mayoral: Laboratorio Central de Veterinaria. Nacional Referente Laboratory for TSEs diagnosis. MAPA. Algete, Madrid, Spain; J. Castilla, C. Soto: Department of Neurology, University of Texas Medical Branch, Galveston, Texas, USA. E-mail: tmayoral@mapya.es
SP englisch
PO Italien