title: Multiplex polymerase chain reaction on FTA cards vs. flow cytometry for B-lymphocyte clonality.
abstract: BACKGROUND
  Two-colour flow cytometry was compared with multiplex PCR with capillary electrophoresis for clonality determination in specific categories of B-cell lymphoma. FTA cards were evaluated for preserving DNA from node imprints and expediting molecular analysis.
  
  
  METHODS
  A single-tube multiplex PCR targeted IGH and lymphoma-specific translocations in DNA extracted from 180 frozen lymphoid tissues and DNA bound to FTA cards from 192 fresh tissues and 137 aspirates. PCR results were compared with flow cytometry in the extracted and aspirated samples.
  
  
  RESULTS
  Overall, single-tube multiplex PCR sensitivity was equivalent in the sample groups (intergroup range 79%-91%). False negatives were associated with tumour origin in the follicle centre. Multiplex PCR and flow cytometry were equally sensitive and together detected 98% of B-cell lymphomas. Additional two-tube targeting of IGK suggested an overall molecular sensitivity >90%. False positive (pseudoclonal) single-tube multiplex PCR was associated with necrosis and sparse lymphocytes.
  
  
  CONCLUSIONS
  Multiplex PCR using template DNA bound to an FTA card effectively detects B-lymphocyte clonality, obviates DNA extraction and refrigeration, and can be used without diminished sensitivity in fine needle aspirates or node imprints as a replacement for or complement to flow cytometry at any point in the diagnostic work-up.
authors: Department of Pathology, Lund University Hospital, Lund, Sweden. michael.dictor@med.lu.se
authors: Michael
authors: M
authors: Dictor
authors: Ingela
authors: I
authors: Skogvall
authors: Janina
authors: J
authors: Warenholt
authors: Eva
authors: E
authors: Rambech
mesh_terms: D001402:B-Lymphocytes / Q000502:physiology*; D044963:Biopsy, Fine-Needle; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D016172:DNA Fingerprinting / Q000379:methods; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D016393:Lymphoma, B-Cell / Q000235:genetics* / Q000473:pathology; D016399:Lymphoma, T-Cell / Q000235:genetics* / Q000473:pathology; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D012189:Retrospective Studies; D012680:Sensitivity and Specificity
chemical_list: D004273:DNA, Neoplasm; D004247:DNA