title: Comparison of two real-time quantitative polymerase chain reaction strategies for minimal residual disease evaluation in lymphoproliferative disorders: correlation between immunoglobulin gene mutation load and real-time quantitative polymerase chain reaction performance.
abstract: We compared two strategies for minimal residual disease evaluation of B-cell lymphoproliferative disorders characterized by a variable immunoglobulin heavy chain (IGH) genes mutation load. Twenty-five samples from chronic lymphocytic leukaemia (n = 18) or mantle cell lymphoma (n = 7) patients were analyzed. Based on IGH variable region genes, 22/25 samples carried > 2% mutations, 20/25 > 5%. In the IGH joining region genes, 23/25 samples carried > 2% mutations, 18/25 > 5%. Real-time quantitative polymerase chain reaction was performed on IGH genes using two strategies: method A utilizes two patient-specific primers, whereas method B employs one patient-specific and one germline primer, with different positions on the variable, diversity and joining regions. Twenty-three samples (92%) resulted evaluable using method A, only six (24%) by method B. Method B poor performance was specifically evident among mutated IGH variable/joining region cases, although no specific mutation load above, which the real-time quantitative polymerase chain reaction failed was found. The molecular strategies for minimal residual disease evaluation should be adapted to the B-cell receptor features of the disease investigated.
authors.affiliation: Department of Cellular Biotechnologies and Hematology, "Sapienza" University, Rome, Italy.
authors.forename: Irene
authors.initials: I
authors.lastname: Della Starza
authors.forename: Marzia
authors.initials: M
authors.lastname: Cavalli
authors.forename: Ilaria
authors.initials: I
authors.lastname: Del Giudice
authors.forename: Daniela
authors.initials: D
authors.lastname: Barbero
authors.forename: Barbara
authors.initials: B
authors.lastname: Mantoan
authors.forename: Elisa
authors.initials: E
authors.lastname: Genuardi
authors.forename: Marina
authors.initials: M
authors.lastname: Urbano
authors.forename: Claudia
authors.initials: C
authors.lastname: Mannu
authors.forename: Anna
authors.initials: A
authors.lastname: Gazzola
authors.forename: Elena
authors.initials: E
authors.lastname: Ciabatti
authors.forename: Anna
authors.initials: A
authors.lastname: Guarini
authors.forename: Robin
authors.initials: R
authors.lastname: Foà
authors.forename: Sara
authors.initials: S
authors.lastname: Galimberti
authors.forename: Pierpaolo
authors.initials: P
authors.lastname: Piccaluga
authors.forename: Gianluca
authors.initials: G
authors.lastname: Gaidano
authors.forename: Marco
authors.initials: M
authors.lastname: Ladetto
authors.forename: Luigia
authors.initials: L
authors.lastname: Monitillo
mesh_terms: D005787:Gene Frequency; D015321:Gene Rearrangement; D005803:Genes, Immunoglobulin; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D008232:Lymphoproliferative Disorders / Q000175:diagnosis* / Q000235:genetics*; D009154:Mutation; D018365:Neoplasm, Residual; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D007143:Immunoglobulin Heavy Chains
keywords: MRD; RQ-PCR; immunoglobulin genes; lymphoproliferative disorders; mutations