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Technical Advances |
From the Clinica di Oncoematologia Pediatrica, Azienda Ospedaliera-Università di Padova, Padova, Italy
The t(8;14)(q24;q32), involving MYC gene (8q24) and the
immunoglobulin heavy chain (IgH) locus (14q32), represents
about 75% of all translocations in Burkitt’s lymphoma (BL). Due to
the great variability of the breakpoint region, a standard
polymerase chain reaction assay is not sufficient for the detection of
this chromosomal translocation. The availability of new and more
efficient DNA polymerases that allow the amplification of genomic
fragments many kilobase-pairs long, makes it possible to identify
the t(8;14) in BL cells by long-distance polymerase chain reaction
(LD-PCR). We have established a simplified and efficient LD-PCR for the
detection of t(8;14)(q24;q32) that relies on the use of one primer
specific for MYC exon II combined, in different
reactions, with four primers for the IgH locus: three for the
constant regions Cµ, C
, and C
, and one
for the joining region (JH). We first studied seven BL cell lines and
optimized LD-PCR reaction for analysis of tumor specimens. Five of
seven cell lines were positive for the t(8;14), whereas two
lines derived from endemic BL were negative, as expected. Of 15
biopsies obtained from pediatric BL and subsequently analyzed,
13 (87%) were positive for the translocation detected by LD-PCR and
showed a product ranging in size from 2.0 to 9.5 kb. Cµ region was
involved in 6 cases, C and C in 2 cases each, and
JH in 3 cases. Interestingly, 2 of the tumors positive for JH
showed a second, larger PCR product with the C- and
C-specific primer, respectively. We established that our
LD-PCR method could detect 10-3 BL cells within a
population of hematopoietic cells lacking the translocation. In
conclusion, our LD-PCR method represents a fast, highly
sensitive, and specific tool to study sporadic BL and to detect
minimal disease and residual disease in patients affected by
t(8;14)-positive lymphomas.
This article has been cited by other articles:
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  L. Mussolin, M. Pillon, V. Conter, M. Piglione, L. Lo Nigro, P. Pierani, C. Micalizzi, S. Buffardi, G. Basso, L. Zanesco, et al. Prognostic Role of Minimal Residual Disease in Mature B-Cell Acute Lymphoblastic Leukemia of Childhood J. Clin. Oncol., November 20, 2007; 25(33): 5254 - 5261. [Abstract] [Full Text] [PDF]
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