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.
Original language | English |
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Pages (from-to) | 364-370 |
Number of pages | 7 |
Journal | Journal of Molecular Diagnostics |
Volume | 8 |
Issue number | 3 |
Publication status | Published - Jul 2006 |
Keywords
- Base Sequence
- Computer Systems
- Consensus
- DNA Probes
- Humans
- Immunoglobulin Heavy Chains
- Molecular Sequence Data
- Multiple Myeloma
- Neoplasm, Residual
- Polymerase Chain Reaction
- Protein Structure, Tertiary