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Validation of BIOMED-2 multiplex PCR tubes for detection of TCRB gene rearrangements in T-cell malignancies

2004; Springer Nature; Volume: 18; Issue: 9 Linguagem: Inglês

10.1038/sj.leu.2403428

1476-5551

J Droese, Anton W. Langerak, Pascal Groenen, Monika Brüggemann, Paul E. Neumann, I L Wolvers-Tettero, M C van Altena, Michael Kneba, Jacques J. M. van Dongen,

Chronic Lymphocytic Leukemia Research

The BIOMED-2 Concerted Action BMH4-CT98-3936 on 'Polymerase chain reaction (PCR)-based clonality studies for early diagnosis of lymphoproliferative disorders' developed standardized PCR protocols for detection of immunoglobulin (Ig) and T-cell receptor (TCR) rearrangements, including TCR beta (TCRB). As no comparable TCRB PCR method pre-existed and only a limited number of samples was tested within the BIOMED-2 study, we initiated this study for further validation of the newly developed TCRB PCR approach by comparing PCR data with previously generated Southern blot (SB) data in a series of 66 immature (ALL) and 36 mature T-cell malignancies. In 91% of cases, concordant PCR and SB results were found. Discrepancies consisted of either failure to detect SB-detected TCRB rearrangements by PCR (6.5%) or detection of an additional non-SB defined rearrangement (2.5%). In 99% of cases (99/100), at least one clonal TCRB rearrangement was detected by PCR in the SB-positive cases. A predominance of complete Vβ-Jβ rearrangements was seen in TCRαβ+ T-cell malignancies and CD3-negative T-ALL (100 and 90%, respectively), whereas in TCRγδ+ T-ALL, more incomplete Dβ-Jβ TCRB rearrangements were detected (73%). Our results underline the reliability of this new TCRB PCR method and its strategic applicability in clonality diagnostics of lymphoproliferative disorders and MRD studies.