title: 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.
abstract: METHODS
  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).
  
  
  METHODS
  The t(2;5) was detected with a long-range nested polymerase chain reaction (PCR) using 0.5 microgram of DNA (60,000-80,000 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 beta-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 10(4)-fold or 10(5)-fold generated amplicons in 100% or 20% of reactions, respectively. Archival tumor DNA from 144 patients was amplifiable by beta-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.
authors.affiliation: Department of Hematology, University of Texas M. D. Anderson Cancer Center, Houston, USA.
authors.forename: A H
authors.initials: AH
authors.lastname: Sarris
authors.forename: R
authors.initials: R
authors.lastname: Luthra
authors.forename: V
authors.initials: V
authors.lastname: Papadimitracopoulou
authors.forename: M
authors.initials: M
authors.lastname: Waasdorp
authors.forename: M A
authors.initials: MA
authors.lastname: Dimopoulos
authors.forename: J A
authors.initials: JA
authors.lastname: McBride
authors.forename: F
authors.initials: F
authors.lastname: Cabanillas
authors.forename: M
authors.initials: M
authors.lastname: Duvic
authors.forename: A
authors.initials: A
authors.lastname: Deisseroth
authors.forename: S W
authors.initials: SW
authors.lastname: Morris
authors.forename: W C
authors.initials: WC
authors.lastname: Pugh
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002889:Chromosomes, Human, Pair 2; D002895:Chromosomes, Human, Pair 5; D005260:Female; D015894:Genome, Human; D006689:Hodgkin Disease / Q000235:genetics*; D006801:Humans; D016403:Lymphoma, Large B-Cell, Diffuse / Q000235:genetics; D008228:Lymphoma, Non-Hodgkin / Q000235:genetics*; D017731:Lymphomatoid Papulosis / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014178:Translocation, Genetic; D014407:Tumor Cells, Cultured