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Immunohistochemistry in an automated platform forPrion Protein in Transmissible Spongiform Encephalopathy
2019 (English)Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
Abstract [en]

Background: Transmissible spongiform encephalopathies, like Creutzfeldt-Jakobs Diseaseare fatal neurodegenerative disorders. They are caused by misfolding and aggregation of anendogenously expressed protein, prion. Detection of the pathological form of prions byimmunohistochemistry has become a valuable tool for diagnostics. However,immunohistochemistry of prions is traditionally performed manually and with various tediouspretreatments.Purpose: In clinical diagnostics fast, specific, reliable and standardised protocols are needed.In this study an optimised protocol for prion immunohistochemistry in an automated platformwas developed.Material and methods: Sections of paraffin-embedded brain tissue from patients with andwithout sporadic Creutzfeldt-Jakobs Disease was used for comparison withimmunohistochemistry with two different anti-prion antibodies 12F10 and 3F4. The effect ofdifferent pre-treatments like vaporised cooking under pressure, use of proteinase K, citratebuffer, guanidine thiocyanate and picric acid was investigated.Results: Both antibodies displayed similar patterns of immunoreactivity in the manual as wellas in the automated platform. Vaporised cooking under pressure was preferred before heatingin the automated platform due to minimal change in tissue morphology with the former. Theuse of citrate buffer and proteinase K was essential for detecting immunoreactivity whereasguanidine thiocyanate and picric acid could be excluded.Conclusion: Both 12F10 and 3F4 can be used for immunohistochemistry on an automatedplatform for clinical diagnostics. Pre-treatments in this protocol include formic acid,vaporised cooking under pressure in citrate buffer and the use of proteinase K.

Place, publisher, year, edition, pages
2019.
Keywords [en]
Creutzfeldt-Jakobs Disease, PrP, IHC, antigen retrieval, neuropathology
National Category
Biomedical Laboratory Science/Technology
Identifiers
URN: urn:nbn:se:uu:diva-377656OAI: oai:DiVA.org:uu-377656DiVA, id: diva2:1291304
External cooperation
Karolinska sjukhuset
Educational program
Biomedical Laboratory Science Programme; Biomedical Laboratory Science Programme
Supervisors
Examiners
Available from: 2019-02-24 Created: 2019-02-24 Last updated: 2019-02-24Bibliographically approved

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CiteExportLink to record
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Citation style
  • apa
  • ieee
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  • de-DE
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  • en-US
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