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From the Division of Pathology,
*
City of Hope National Medical Center, Duarte, California; the Department of Pathology,
Oregon Health Sciences University, Portland, Oregon; the Department of Pathology and Laboratory Medicine,
University of Pennsylvania, Philadelphia, Pennsylvania; and the Department of Cellular Pathology,
Armed Forces Institute of Pathology, Washington, DC
To evaluate current diagnostic methods used for the evaluation of T
cell receptor (TCR) gene rearrangements, 24 different
laboratories analyzed 29 lymphoid neoplasm samples of extracted DNA and
paraffin-embedded tissue and were asked to complete a technical
questionnaire related to the testing. Participating laboratories
performed Southern blot and polymerase chain reaction (PCR) testing for
rearrangements of the TCRß chain gene and PCR for
the TCR
chain gene rearrangements. Of 14 laboratories
performing TCRß Southern blot analysis, there was
complete agreement in 10 of 14 cases, with some false negative
results obtained in 4 cases. No false positive results were obtained by
Southern blot analysis. TCRß PCR analysis was only
performed by two laboratories, and only 47.1% of positive
samples were detected. Twenty-one laboratory results were obtained for
TCR PCR. This method showed an overall detection rate of
77.9% for T cell gene rearrangements with a 4.1% false positive
rate, as compared to both TCR Southern blot
analysis results and immunophenotyping. The detection rate for
TCR PCR, however, significantly differed
when extracted DNA samples from frozen tissue were compared to
paraffin-embedded tissue (85.4% versus 65.9%;
P = 0.0005). Significant differences in true
positive results were obtained when laboratories using primers directed
against multiple TCR variable regions (V1–8 plus one to
three other primer sets) were compared to laboratories that used only a
single set of TCR primers directed against the V1–8
(P < 0.0001). Other technical factors
significantly affecting results were also identified. These findings
provide useful data on the current state of diagnostic TCR
testing, highlight the risk of false negative results for TCR
testing directed against only portions of the TCR
gene, and identify limitations of testing of paraffin-embedded
tissues in some laboratories.
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