Abstract
BackgroundFormalin-fixed paraffin-embedded tissue is regularly employed in the diagnosis of transmissible spongiform encephalopathies (TSE) by immunohistochemistry (IHC), the standard by which all other TSE diagnostic protocols are 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 analyzing individual sections by hand does not allow for rapid or high throughput screening. However, preservation of tissues in formalin is not dependent upon the availability of refrigeration.FindingsFormalin-fixed paraffin-embedded tissues from TSE transmission studies of scrapie in sheep, chronic wasting disease in white-tailed deer or transmissible mink encephalopathy in cattle were cut at 5 μm thickness. Samples containing the tissue equivalent of as little as one 5 μm section can be used to readily discriminate positive from negative samples.ConclusionsThis approach cannot replace IHC but may be used along with IHC as both a more rapid and readily high throughput screen where fresh or frozen tissues are not available or impractical.
Highlights
Formalin-fixed paraffin-embedded tissue is regularly employed in the diagnosis of transmissible spongiform encephalopathies (TSE) by immunohistochemistry (IHC), the standard by which all other TSE diagnostic protocols are judged
Due to the lack of a defined immune response or nucleic acid component of the infectious agent, approaches for transmissible spongiform encephalopathy (TSE) diagnosis rely upon methods of immunodetection including immunohistochemistry (IHC), Western blotting and enzymelinked immunosorbent assay (ELISA)-based approaches for detection of the infectious agent. [1,2,3] Generally speaking, IHC relies upon formalin fixed paraffin embedded tissues, while Western blotting and ELISA utilize fresh or frozen tissues
Sample preparation The method described here is an extension of previously published methods for Western blotting of formalin-fixed paraffin embedded samples differing only in the method for detecting PrPSc [4,5]
Summary
Formalin-fixed paraffin-embedded tissue is regularly employed in the diagnosis of transmissible spongiform encephalopathies (TSE) by immunohistochemistry (IHC), the standard by which all other TSE diagnostic protocols are judged. [1,2,3] Generally speaking, IHC relies upon formalin fixed paraffin embedded tissues, while Western blotting and ELISA utilize fresh or frozen tissues. Methods have been reported that allow detection of PrPSc in formalin fixed tissues by Western blot [4,5,6].
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
