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Analysis of T cell receptor beta chain CDR3 size using RNA extracted from formalin fixed paraffin wax embedded tissue.

AbstractAIMS:
To isolate RNA and DNA simultaneously from formalin fixed paraffin wax embedded tissue to assess the clonality of enteropathy associated T cell lymphomas and to analyse it in detail by a non-radioactive method of T cell receptor complementarity determining region 3 (CDR3) spectratyping.
METHODS:
DNA and RNA were isolated simultaneously from formalin fixed paraffin wax embedded tissue blocks and subjected to the polymerase chain reaction (PCR) and semi-nested reverse transcription PCR (RT-PCR), respectively. The RT-PCR T cell receptor V beta products were analysed by CDR3 spectratyping using a denaturing polyacrylamide gel and silver staining.
RESULTS:
Usable DNA and RNA were isolated simultaneously from formalin fixed paraffin wax embedded tissue. The specific clonality of the tissue was successfully analysed by a non-radioactive method of T cell receptor CDR3 spectratyping of the RT-PCR products. CDR3 spectratying of the RT-PCR products demonstrated the precise clonal nature of the tumour and non-tumour tissue showing that the non-tumour tissue comprised an oligoclonal population of a number of different T cell receptor V beta families. The tumour tissue comprised two T cell subtypes of the one family, T cell receptor V beta 9.
CONCLUSIONS:
RNA and DNA were isolated from formalin fixed paraffin wax embedded enteropathy associated T cell lymphoma tissue. Detailed analysis of clonality can be carried out by a non-radioactive method of CDR3 spectratyping.
AuthorsU O'Shea, J I Wyatt, P D Howdle
JournalJournal of clinical pathology (J Clin Pathol) Vol. 50 Issue 10 Pg. 811-4 (Oct 1997) ISSN: 0021-9746 [Print] England
PMID9462260 (Publication Type: Journal Article)
Chemical References
  • DNA, Neoplasm
  • RNA, Neoplasm
  • Receptor-CD3 Complex, Antigen, T-Cell
  • Formaldehyde
Topics
  • DNA, Neoplasm (analysis)
  • Electrophoresis, Polyacrylamide Gel
  • Formaldehyde
  • Humans
  • Lymphoma, T-Cell (genetics)
  • Paraffin Embedding
  • Polymerase Chain Reaction (methods)
  • RNA, Neoplasm (analysis)
  • Receptor-CD3 Complex, Antigen, T-Cell (analysis)
  • Silver Staining

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