anaplastic large-cell lymphoma
genomic DNA PCR
Springer Online Journal Archives 1860-2000
Abstract 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 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 or105-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 who 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: PCR of genomic DNA detects t(2;5) only in ALCL, but not in other NHLs, HD, or LP, and may be useful in establishing diagnosis, determining prognosis, and monitoring minimal residual disease in ALCL.
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