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

J. Martinez-Lopez, P. Martinez-Sanchez, R. Garcia-Sanz, M.E. Sarasquete, R. Ayala, M. Gonzalez, J.M. Bautista, D. Gonzalez, J. San Miguel, G. Garcia-Effron, J.J. Lahuerta

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2 Citations (Scopus)

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 languageEnglish
Pages (from-to)364-370
Number of pages7
JournalJournal of Molecular Diagnostics
Volume8
Issue number3
Publication statusPublished - 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

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    Martinez-Lopez, J., Martinez-Sanchez, P., Garcia-Sanz, R., Sarasquete, M. E., Ayala, R., Gonzalez, M., Bautista, J. M., Gonzalez, D., San Miguel, J., Garcia-Effron, G., & Lahuerta, J. J. (2006). Application of self-quenched JH consensus primers for real-time quantitative PCR of IGH gene to minimal residual disease evaluation in multiple myeloma. Journal of Molecular Diagnostics, 8(3), 364-370. http://www.ncbi.nlm.nih.gov/pubmed/16825510