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Blood, Vol. 96 No. 2 (July 15), 2000:
pp. 640-646
High detection rate of T-cell receptor beta chain rearrangements
in T-cell lymphoproliferations by family specific polymerase chain
reaction in combination with the GeneScan technique and DNA sequencing
Chalid Assaf,
Michael Hummel,
Edgar Dippel,
Sergij Goerdt,
Hans-Henning Müller,
Ioannis Anagnostopoulos,
Constantin E. Orfanos, and
Harald Stein
From the Institute of Pathology, Consultation and Reference Center
for Lymph Node Pathology and Hematopathology and Department of
Dermatology, University Medical Centre Benjamin Franklin, The Free
University of Berlin, Berlin, Germany.
The distinction between benign polyclonal and malignant monoclonal
lymphoid disorders by morphology or immunophenotyping is frequently
difficult. Therefore, the demonstration of clonal B-cell or T-cell
populations by detecting identically rearranged immunoglobulin (Ig) or
T-cell receptor (TCR) genes is often used to solve this diagnostic
problem. Whereas the detection of rearranged Ig genes is well
established, TCR gamma ( ) and beta ( ) gene rearrangements often
escape detection with the currently available polymerase chain reaction
(PCR) assays. To establish a sensitive, specific, and rapid method for
the detection of rearranged TCR- genes, we developed a new PCR
approach with family-specific J primers and analyzed the resulting
PCR products by high-resolution GeneScan technique. The superior
efficiency of this new method was demonstrated by investigating 132 DNA
samples extracted from lymph node and skin biopsy specimens (mostly
formalin fixed) and blood samples of 62 patients who had a variety of
T-cell lymphomas and leukemias. In all but 1 of the tumor samples
(98.4%) a clonal amplificate was detectable after TCR- PCR and the
same clonal T-cell population was also found in 15 of 18 (83%) of the
regional lymph nodes and in 7 of 11 (64%) of the peripheral blood
samples. Direct comparison of these results with those obtained
currently by the most widely applied TCR- PCR revealed an
approximate 20% lower detection rate in the same set of samples than
with the TCR- PCR method. These results indicate that the new
TCR- PCR is most suitable for a rapid and reliable detection of
clonal T-cell populations.
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