title: Application of self-quenched JH consensus primers for real-time quantitative PCR of IGH gene to minimal residual disease evaluation in multiple myeloma.
abstract: Monitoring multiple myeloma patients for relapse requires sensitive methods to measure minimal residual disease and to establish a more precise prognosis. The present study aimed to standardize a real-time quantitative polymerase chain reaction (PCR) test for the IgH gene with a JH consensus self-quenched fluorescence reverse primer and a VDJH or DJH allele-specific sense primer (self-quenched PCR). This method was compared with allele-specific real-time quantitative PCR test for the IgH gene using a TaqMan probe and a JH consensus primer (TaqMan PCR). We studied nine multiple myeloma patients from the Spanish group treated with the MM2000 therapeutic protocol. Self-quenched PCR demonstrated sensitivity of >or=10(-4) or 16 genomes in most cases, efficiency was 1.71 to 2.14, and intra-assay and interassay reproducibilities were 1.18 and 0.75%, respectively. Sensitivity, efficiency, and residual disease detection were similar with both PCR methods. TaqMan PCR failed in one case because of a mutation in the JH primer binding site, and self-quenched PCR worked well in this case. In conclusion, self-quenched PCR is a sensitive and reproducible method for quantifying residual disease in multiple myeloma patients; it yields similar results to TaqMan PCR and may be more effective than the latter when somatic mutations are present in the JH intronic primer binding site.
authors.affiliation: Servicio de Hematología, Hospital Universitario, 12 de Octubre. Av. de Cordoba, s/n Madrid 28041, Spain. mpmartinezsa@yahoo.es
authors.forename: Joaquin
authors.initials: J
authors.lastname: Martinez-Lopez
authors.forename: Pilar
authors.initials: P
authors.lastname: Martinez-Sanchez
authors.forename: Ramon
authors.initials: R
authors.lastname: Garcia-Sanz
authors.forename: Maria Eugenia
authors.initials: ME
authors.lastname: Sarasquete
authors.forename: Rosa
authors.initials: R
authors.lastname: Ayala
authors.forename: Marcos
authors.initials: M
authors.lastname: Gonzalez
authors.forename: Jose Manuel
authors.initials: JM
authors.lastname: Bautista
authors.forename: David
authors.initials: D
authors.lastname: Gonzalez
authors.forename: Jesus
authors.initials: J
authors.lastname: San Miguel
authors.forename: Guillermo
authors.initials: G
authors.lastname: Garcia-Effron
authors.forename: Juan Jose
authors.initials: JJ
authors.lastname: Lahuerta
mesh_terms: D001483:Base Sequence; D003199:Computer Systems; D032921:Consensus; D015342:DNA Probes; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D008969:Molecular Sequence Data; D009101:Multiple Myeloma / Q000175:diagnosis* / Q000235:genetics*; D018365:Neoplasm, Residual / Q000175:diagnosis* / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D017434:Protein Structure, Tertiary / Q000235:genetics
chemical_list: D015342:DNA Probes; D007143:Immunoglobulin Heavy Chains