Comparative investigations of T-cell receptor 
gene rearrangements in frozen and paraffin-embedded tissue
by capillary electrophoresis
Mariann Christensen 1*, Anette D Funder 1, Knud Bendix 1 and Flemming B Soerensen 1
1 University Institute of Pathology, Aarhus Hospital, Denmark
* To whom correspondence should be addressed. E-mail: lab10mc{at}as.aaa.dk.
Accepted 25 July 2005
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Abstract |
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Aims: The aims of this study were to investigate
clonal TCR 
gene rearrangements in frozen and
paraffin-embedded tissue (FFPE), using capillary
electrophoresis (GeneScan analysis and sequencing, ABI
310 Genetic Analyser) for the use in a diagnostic setting,
since T-cell lymphomas may be difficult to diagnose by
conventional methods.
Methods: The DNA for PCR was extracted from
frozen and FFPE tissue, cell lines and blood. PCR primers:
V 1-8, V 9, V 10 or V 11
(5'end labelled) combined with a mixture of J P1/J
P/J P2/J 2 (unlabelled).
Monoclonal cases were sequenced. The following analyses
were performed: clonality, reproducibility, sensitivity
and specificity.
Results: In all cases the molecular test was in
agreement with the histological diagnosis. Discrepancies
were found between frozen and FFPE tissue in 18 out of 56
tests (32%). The method was highly reproducible. The
sensitivity was 0.5 % for cell lines and 1.0 % for patient
specimens. The specificity was 100%. The junctional region
between the V and J segments was
specific for individual patients.
Conclusions: Capillary electrophoresis of PCR
products from frozen and FFPE tissue is suitable for
detecting clonal TCRgene rearrangements. However,
it is of importance to correlate the results with
conventional morphological and immunohistochemical studies.
Key Words:
T-cell receptor, capillary electrophoresis, formalin-fixed, paraffin-embedded tissue (FFPE), frozen tissue, gene rearrangement
