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Leukemia. 2014 Feb;28(2):391-7. doi: 10.1038/leu.2013.217. Epub 2013 Jul 17.

Critical evaluation of ASO RQ-PCR for minimal residual disease evaluation in multiple myeloma. A comparative analysis with flow cytometry.

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Department of Hematology, Hospital Universitario de Salamanca; IBSAL; IBMCC (USAL-CSIC), Salamanca, Spain.
Department of Hematology, Hospital 12 de Octubre, Madrid, Spain.
Department of Hematology, Hospital Germans Trias I Pujol, Badalona, Spain.
Department of Hematology, Hospital Clínico 'Lozano Blesa', Zaragoza, Spain.
Department of Hematology, Hospital Universitario La Fe and Universidad Católica 'San Vicente Mártir', Valencia, Spain.
Department of Hematology, Hospital Clinic I Provincial, Barcelona, Spain.


We have analyzed the applicability, sensitivity and prognostic value of allele-specific oligonucleotide real-time quantitative PCR (ASO RQ-PCR) as a method for minimal residual disease (MRD) assessment in patients with multiple myeloma (MM), comparing the results with those of multiparameter flow cytometry (MFC). A total of 170 patients enrolled in three consecutive Spanish trials achieving at least partial response after treatment were included. Lack of clonality detection (n=31), unsuccessful sequencing (n=17) and suboptimal ASO performance (n=51) limited the applicability of PCR to 42% of cases. MRD was finally investigated in 103 patients (including 32 previously studied) with persistent disease identified by PCR and MFC in 54% and 46% of cases, respectively. A significant correlation in MRD quantitation by both the techniques was noted (r=0.881, P<0.001), being reflective of treatment intensity. Patients with <10(-4) residual tumor cells showed longer progression-free survival (PFS) compared with the rest (not reached (NR) vs 31 months, P=0.002), with similar results observed with MFC. Among complete responders (n=62), PCR discriminated two risk groups with different PFS (49 vs 26 months, P=0.001) and overall survival (NR vs 60 months, P=0.008). Thus, although less applicable than MFC, ASO RQ-PCR is a powerful technique to assess treatment efficacy and risk stratification in MM.

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