Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 6/15/2010
Publication Date: 9/8/2010
Citation: Nicholson, E.M. 2010. Formalin fixed paraffin embedded tissue as a starting point for PrPSc detection by ELISA [abstract]. Prion. 4(3):213.
Technical Abstract: Introduction: Formalin fixed paraffin embedded tissue are regularly employed in TSE diagnosis by IHC, the standard by which all other diagnostic protocols are currently judged. While IHC affords advantages over diagnostic approaches that typically utilize fresh or frozen tissue, such as Western blot and ELISA, the process of fixing, staining, and analysis of individual sections by hand does not readily allow for either rapid nor high through put screening. However, preservation of tissues in formalin is not dependent upon the availability of refrigeration. Materials and Methods: Formalin fixed paraffin embedded tissues resulting from TSE transmission studies at the National Animal Disease Center were cut at 4µm thickness. The samples were boiled to allow manual removal of paraffin, sonicated to disrupt the tissue and analyzed using a commercial ELISA test kit. Results: Samples containing as little as one 4 µm section can be used to readily discrimiate positive from negative samples. Discussion: This approach cannot replace IHC but may be used with IHC approach as both a more rapid and readily high through put screen where fresh or frozen tissues are not available or not practical. Applicability to different TSEs and time in formalin are important considerations and are discussed.