1. Detection of translocations affecting the BCL6 locus in B cell non-Hodgkin's lymphoma by interphase fluorescence in situ hybridization.
- Author
-
Sanchez-Izquierdo, D, Siebert, R, Harder, L, Marugan, I, Gozzetti, A, Price, H P, Gesk, S, Hernandez-Rivas, J M, Benet, I, Solé, F, Sonoki, T, Le Beau, M M, Schlegelberger, B, Dyer, M J S, Garcia-Conde, J, Martinez-Climent, J A, and Dyer, M J
- Subjects
CHROMOSOMAL translocation ,B cells ,LYMPHOMAS ,B cell lymphoma ,CHROMOSOME abnormalities ,CHROMOSOME banding ,CHROMOSOMES ,COMPARATIVE studies ,DOCUMENTATION ,GENES ,KARYOTYPES ,RESEARCH methodology ,MEDICAL cooperation ,NUCLEOTIDES ,RESEARCH ,FLUORESCENCE in situ hybridization ,EVALUATION research - Abstract
Structural alterations in 3q27 affecting the BCL6 locus are among the most frequent changes in B-NHL. The aim of the present study was to establish an interphase-FISH assay for the detection of all diverse BCL6 translocations in B-NHL. Two different approaches were tested, one using a PAC-clone spanning the major breakpoint region (MBR) of BCL6 (span-assay), and another using two BAC clones flanking the MBR (flank-assay). Interphase FISH with the span-assay detected the various BCL6 translocations in seven B-NHL cell lines. The dual-color flank-assay was evaluated in two laboratories independently: in normal controls, the cutoff level for false-positive signals was 2.6%, whereas the cutoff level for false-negatives in the seven cell lines was 7.5%. To test the feasibility of the FISH strategies, 30 samples from patients with B-NHL with cytogenetic abnormalities of 3q27 were evaluated with both assays. In 21 cases, the span-assay indicated a BCL6 rearrangement. In 18 of the 21 cases, the dual-color flank-assay confirmed the translocation including 12 different partner chromosomal loci. The three false-positive cases detected with the span-assay showed trisomy of chromosome 3 by cytogenetic analyses, and they were correctly classified as non-rearranged with the flank-assay. In summary, our FISH strategy using two differently labeled flanking BCL6 BAC probes provides a robust, sensitive, and reproducible method for the detection of common and uncommon abnormalities of BCL6 gene in interphase nuclei. The routine application of this assay to patients with B-NHL will allow the assessment of the diagnostic and prognostic significance of BCL6 rearrangements. [ABSTRACT FROM AUTHOR]
- Published
- 2001
- Full Text
- View/download PDF