title: A practical strategy for the routine use of BIOMED-2 PCR assays for detection of B- and T-cell clonality in diagnostic haematopathology.
abstract: BIOMED-2 polymerase chain reaction (PCR) assays for clonality analysis of immunoglobulin (IG) and T-cell receptor (TCR) gene rearrangements were evaluated in routine haematopathological practice where paraffin-embedded tissues constitute the majority of specimens. One hundred and twenty-five fresh/frozen and 316 paraffin specimens were analysed for DNA quality and clonality. Seventy-nine per cent of paraffin specimens yielded PCR products of over 300 bp. These specimens and all fresh/frozen specimens were analysed with the complete set of BIOMED-2 reactions for IG (8 reactions) and/or TCR (6 reactions) gene rearrangements. The rate of detection of clonality was 96% in mature B-cell neoplasms and 98% in mature T-cell neoplasms and there were no significant differences in these rates between paraffin and fresh/frozen specimens. As the value of sole use of any individual BIOMED-2 reaction in clonality detection was limited, we assessed combinations of reactions that gave the greatest sensitivity with fewest reactions and were applicable for both fresh/frozen and paraffin specimens. For IG gene rearrangements, three reactions combining one targeting the IG heavy chain framework-2 region and two targeting the IG kappa locus achieved a 91% detection rate. For TCR gene rearrangements, the two TCR gamma reactions gave a 94% detection rate. We therefore recommend this strategy as the first-line assays for routine B- and T-cell clonality analysis in diagnostic haematopathology.
authors.affiliation: Department of Histopathology, Addenbrooke's Hospital, Cambridge, UK.
authors.forename: Hongxiang
authors.initials: H
authors.lastname: Liu
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Bench
authors.forename: Chris M
authors.initials: CM
authors.lastname: Bacon
authors.forename: Karen
authors.initials: K
authors.lastname: Payne
authors.forename: Yuanxue
authors.initials: Y
authors.lastname: Huang
authors.forename: Mike A
authors.initials: MA
authors.lastname: Scott
authors.forename: Wendy N
authors.initials: WN
authors.lastname: Erber
authors.forename: John W
authors.initials: JW
authors.lastname: Grant
authors.forename: Ming-Qing
authors.initials: MQ
authors.lastname: Du
mesh_terms: D001402:B-Lymphocytes / Q000473:pathology*; D002999:Clone Cells; D017931:DNA Primers / Q000235:genetics; D001330:Electronic Data Processing; D015321:Gene Rearrangement; D019337:Hematologic Neoplasms / Q000175:diagnosis* / Q000235:genetics; D006801:Humans; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000473:pathology*
chemical_list: D017931:DNA Primers