title: BIOMED-2 multiplex immunoglobulin/T-cell receptor polymerase chain reaction protocols can reliably replace Southern blot analysis in routine clonality diagnostics.
abstract: To establish the most sensitive and efficient strategy of clonality diagnostics via immunoglobulin and T-cell receptor gene rearrangement studies in suspected lymphoproliferative disorders, we evaluated 300 samples (from 218 patients) submitted consecutively for routine diagnostics. All samples were studied using the BIOMED-2 multiplex polymerase chain reaction (PCR) protocol. In 176 samples, Southern blot (SB) data were also available, and the two types of molecular results were compared. Results of PCR and SB analysis of both T-cell receptor and immunoglobulin loci were concordant in 85% of samples. For discordant results, PCR results were more consistent with the final diagnosis in 73% of samples. No false-negative results were obtained by PCR analysis. In contrast, SB analysis failed to detect clonality in a relatively high number of samples, mainly in cases of low tumor burden. We conclude that the novel BIOMED-2 multiplex PCR strategy is of great value in diagnosing patients with suspected B- and T-cell proliferations. Because of its higher speed, efficiency, and sensitivity, it can reliably replace SB analysis in clonality diagnostics in a routine laboratory setting. Just as with SB results, PCR results should always be interpreted in the context of clinical, immunophenotypical, and histopathological data.
authors.affiliation: Department of Immunology, Erasmus MC, University Medical Center Rotterdam, Dr. Molewaterplein 50, 3015 GE Rotterdam, The Netherlands.
authors.forename: Yorick
authors.initials: Y
authors.lastname: Sandberg
authors.forename: Ellen J
authors.initials: EJ
authors.lastname: van Gastel-Mol
authors.forename: Brenda
authors.initials: B
authors.lastname: Verhaaf
authors.forename: King H
authors.initials: KH
authors.lastname: Lam
authors.forename: Jacques J M
authors.initials: JJ
authors.lastname: van Dongen
authors.forename: Anton W
authors.initials: AW
authors.lastname: Langerak
mesh_terms: D001402:B-Lymphocytes / Q000166:cytology / Q000378:metabolism; D015139:Blotting, Southern; D049109:Cell Proliferation; D002999:Clone Cells / Q000378:metabolism* / Q000473:pathology*; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics*; D008232:Lymphoproliferative Disorders / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D016133:Polymerase Chain Reaction; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*
chemical_list: D007136:Immunoglobulins; D011948:Receptors, Antigen, T-Cell