Histopathology. 2006 Mar;48(4):363-76.
Ewing's sarcoma of bone: the detection of specific transcripts
in a large, consecutive series of formalin-fixed, decalcified,
paraffin-embedded tissue samples using the reverse
transcriptase-polymerase chain reaction.
Mangham DC, Williams A, McMullan DJ, McClure J, Sumathi VP, Grimer RJ,
Department of Musculoskeletal Pathology, Royal Orthopaedic Hospital,
The Medical School, University of Birmingham, Birmingham, UK.
Aims : (i) To report on the routine use of the reverse
transcriptase-polymerase chain reaction (RT-PCR) technique on
decalcified or non-decalcified, formalin-fixed, paraffin-embedded
tissue (FFPET) for translocation detection, with particular emphasis on
improved RNA extraction methodology and the use of PCR primers designed
to generate small amplicons. (ii) To report on the relative incidences
of translocation types and transcript variants in a large, single
institution series of Ewing's sarcoma of bone. Methods and results :
Using RT-PCR to detect specific transcript variants, we analysed FFPET
from 54 consecutive cases of Ewing's sarcoma of bone. We used 'gold
standard' detection methods on corresponding fresh and fresh frozen
tissue to validate the technique. We have demonstrated the effective
use of RT-PCR on decalcified and non-decalcified FFPET samples for
sarcoma-specific translocation detection (96% sensitivity, 100%
specificity). Tissue decalcification did not affect the detection rate.
The relative incidence of Ewing's sarcoma-specific translocation types
and transcript variants was entirely consistent with previously
published data. Conclusions : With equal effectiveness, RT-PCR can be
applied to both acid decalcified and non-decalcified FFPET for (Ewing's
sarcoma) translocation detection and the technique can be introduced
into routine practice in histopathology departments.
PMID: 16487358 [PubMed – in process]