Evaluation of T cell receptor testing in lympoid neoplasms: Results of a Multicenter Study of 29 Extracted DNA and Paraffin-Embedded Samples

Daniel A. Arber, Rita M. Braziel, Adam Bagg, Karen E. Bijwaard

Research output: Contribution to journalArticle

35 Scopus citations

Abstract

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 TCRβ chain gene and PCR for the TCRγ chain gene rearrangements. Of 14 laboratories performing TCRβ 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. TCRβ PCR analysis was only performed by two laboratories, and only 47.1% of positive samples were detected. Twenty-one laboratory results were obtained for TCRγ 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 TCRγ Southern blot analysis results and immunophenotyping. The detection rate for TCRγ 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 TCRγ 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 TCRγ gene, and identify limitations of testing of paraffin-embedded tissues in some laboratories.

Original languageEnglish (US)
Pages (from-to)133-140
Number of pages8
JournalJournal of Molecular Diagnostics
Volume3
Issue number4
DOIs
StatePublished - Nov 2001

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Molecular Medicine

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