Annals of Oncology 8:S59-S63, 1997
© 1997 European Society for Medical Oncology
Long-range amplification of genomic DNA detects the t(2;5)(p23;q35) in anaplastic large-cell lymphoma, but not in other non-Hodgkin's lymphomas, Hodgkin's disease, or lymphomatoid papulosis
1 Department of Hematology, University of Texas M. D. Anderson Cancer Center Houston, TX
2 Department of Pathology, University of Texas M. D. Anderson Cancer Center Houston, TX
3 Department of Dermatology, University of Texas M. D. Anderson Cancer Center Houston, TX
4 Department of Experimental Oncology, St. Jude Children's Research Hospital Memphis, TN, USA
Correspondence to: Andreas H. Sarris, MD, PhD Department of Hematology, Section of Lymphoma Box 68 University of Texas M. D. Anderson Cancer Center 1515 Holcombe Blvd Houston, TX 77030
Design Determine the frequency of t(2;5)(p23;q35) in anaplastic large-cell lymphoma (ALCL), non-Hodgkin's lymphoma (NHL), Hodgkin's disease (HD), and lymphomatoid papulosis (LP).
Patients and methods The t(2;5) was detected with a long-range nested polymerase chain reaction (PCR) using 0.5 µg of DNA (60000–80000 cells), 5'-primers from the NPM gene, 3'-primers from the ALK gene, agarose electrophoresis, hybridization, and autoradiography. Patients were evaluable if a 3016 base pair amplicon could be generated from tumor DNA with β-globin primers.
Results Amplicons were detected by PCR of genomic DNA from three ALCL cell lines and four primary ALCLs known to t(2;5) positive. DNA from t(2;5)-positive cell lines diluted 104-fold or 105-fold generated amplicons in 100% or 20% of reactions, respectively. Archival tumor DNA from 144 patients was amplifiable by β-globin amplicons in 126 (88%) who are considered evaluable for this study. Twenty-two had ALCL, 69 other NHLs, 30 HD, and five LP. Genomic DNA PCR detected the t(2;5) in 5 of 22 with ALCL (23%, 95% confidence intervals [95% CI] 8%–45%) but not in those with NHLs, HD, or LP. Among ALCLs the t(2;5) was confined to 5 of 20 with nodal presentations (25%, 95% CI 9%–49%), among whom it was seen in 5 of 15 with T-cell or null-cell phenotype (33%, 95% CI 12%–62%), in 4 of 11 with age <40 years (36%, 95% CI 11%–69%), and in 4 of 9 with nodal presentations, T-cell or null-cell phenotype, and age < 40 years (44%, 95% CI 14%–79%). Amplicon sizes were different between cell lines and patients, reflecting unique genomic DNA breakpoints, as confirmed by DNA sequencing, and served as an internal control against specimen cross-contamination in the laboratory.
Conclusions Long-range PCR of genomic DNA detects t(2;5) only in ALCL, but not in other NHLs, HD, or LP. Long-range PCR may be useful in establishing diagnosis, determining prognosis, and monitoring minimal residual disease in ALCL.
anaplastic large-cell lymphoma, genomic DNA, long-range PCR, t(2;5)(p23;q35)