Application of self-quenched JH consensus primers for real-time quantitative PCR of IgH gene to minimal residual disease evaluation in multiple myeloma

MARTÍNEZ-LÓPEZ, JOAQUÍN; MARTÍNEZ-SÁNCHEZ, PILAR; GARCÍA-SANZ, RAMÓN; SARASQUETE, MARÍA EUGENIA; AYALA, ROSA; GONZÁLEZ, MARCOS; BAUTISTA, JOSÉ MANUEL; GONZÁLEZ, DAVID; SAN MIGUEL, JESÚS; GARCÍA-EFFRON, GUILLERMO; LAHUERTA, JUAN JOSÉ

The Journal of molecular diagnostics

American Society for Investigative Pathology y Association for Molecular Pathology

Lugar: Bethesda, MD (EEUU); Año: 2006 vol. 8 p. 364 - 370

1525-1578

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.