NR AOSF
AU Estey,L.; Velek,K.; Koller,S.; Plourde,L.; Toomik,R.; Leathers,V.; Wong,C.; Wilson,S.; Stanley,C.; Tonelli,Q.
TI A Simple and Rapid post mortem EIA Assay for the Detection of Transmissible Spongiform Encephalopathies
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - IV-07
PT Konferenz-Poster
AB
Misfolding of the normal host prion protein (PrPc) and its subsequent accumulation as a protease-resistant conformer (PrPsc) is a well-documented correlate of transmissible spongiform encephalopathies (TSEs). Proteinase K resistance is commonly leveraged as a method for distinguishing PrPsc from PrPc in most TSE diagnostics on the market. IDEXX has developed a TSE diagnostic that does not require proteinase K digestion, but instead uses Seprion capture technology applied to a microtiter plate format. This method utilizes a non-biological PrPsc-specific ligand that selectively binds PrPsc in the presence of excess PrPc. Captured PrPsc is then detected using an anti PrP antibody-HRP conjugate. Sample preparation is limited to homogenization of tissue and the addition of diluent; no other processing is required before applying samples to the assay plate. Assay run time is 3-4 hours, depending on the tissue type under evaluation.
The IDEXX assay has been used to detect PrPsc in characterized samples from bovine and ovine brains as well as cervid lymph nodes. The specificity of the IDEXX assay was 100% in all species and tissues tested. Sensitivity was greater than 99% for non-obex tissue from bovines (versus immunohistochemistry (IHC) conducted on obex tissue) and 100% for a small population of IHC-positive cervid lymph nodes. A 100% correlation between the IDEXX assay and an EU-approved BSE assay was observed for bovine samples. Field evaluations are continuing to test IDEXX kit performance on large populations of bovine, scrapie and cervid samples. The IDEXX assay, with its absence of a proteinase K digestion step and minimal handling during sample preparation, provides a sensitive, rapid and easy to use method for identifying TSE positive samples. The simplicity of the method allows straightforward adaptation to automation, making it an ideal tool for screening large numbers of samples.
AD Lisa Estey, Katherine Velek, Sherri Koller, Lori Plourde, Valerie Leathers, Cai'ne Wong, Quentin Tonelli, IDEXX Laboratories, USA; Reet Toomik, IDEXX Scandinavia AB, Sweden; Stuart Wilson, Christopher Stanley, Microsens Biotechnologies, UK
SP englisch
PO Deutschland