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Blood Cancer J. 2014 Oct 10;4:e250. doi: 10.1038/bcj.2014.69.

Comparative analysis of minimal residual disease detection by multiparameter flow cytometry and enhanced ASO RQ-PCR in multiple myeloma.

Author information

1
1] Department of Medicine, Tampere University Hospital, Tampere, Finland [2] Department of Medicine and Hematology, Kuopio University Hospital, Kuopio, Finland.
2
TYKSLAB, Laboratory of Molecular Genetics, Turku University Hospital, Turku, Finland.
3
1] Fimlab Medical Laboratories Ltd, Tampere University Hospital, Tampere, Finland [2] Department of Clinical Chemistry, Turku University, Turku, Finland.
4
Department of Internal Medicine, Turku University Hospital, Turku University, Turku, Finland.
5
Helsinki University Hospital, Helsinki, Finland.
6
1] NordLab Oulu, Oulu University Hospital, Oulu, Finland [2] Medical Research Center Oulu, Oulu University Hospital and University of Oulu, Oulu, Finland.
7
Laboratory of Eastern Finland, Kuopio University Hospital, Kuopio, Finland.
8
HUSLAB, Laboratory of Clinical Chemistry, Helsinki University Hospital, Helsinki, Finland.
9
TYKSLAB, Laboratory of Clinical Haematology, Turku University Hospital, Turku, Finland.
10
Science Services Center, Kuopio University Hospital, Kuopio, Finland.

Abstract

Multiparameter flow cytometry (MFC) and allele-specific oligonucleotide real-time quantitative PCR (ASO RQ-PCR) are the two most sensitive methods to detect minimal residual disease (MRD) in multiple myeloma (MM). We compared these methods in 129 paired post-therapy samples from 22 unselected, consecutive MM patients in complete/near complete remission. Appropriate immunophenotypic and ASO RQ-PCR-MRD targets could be detected and MRD analyses constructed for all patients. The high PCR coverage could be achieved by gradual widening of the primer sets used for clonality detection. In addition, for 13 (55%) of the patients, reverse orientation of the ASO primer and individual design of the TaqMan probe improved the sensitivity and specificity of ASO RQ-PCR analysis. A significant nonlinear correlation prevailed between MFC-MRD and PCR-MRD when both were positive. Discordance between the methods was found in 32 (35%) paired samples, which were negative by MFC-MRD, but positive by ASO RQ-PCR. The findings suggest that with the described technique, ASO RQ-PCR can be constructed for all patients with MM. ASO RQ-PCR is slightly more sensitive in MRD detection than 6-10-color flow cytometry. Owing to technical demands ASO RQ-PCR could be reserved for patients in immunophenotypic remission, especially in efficacy comparisons between different drugs and treatment modalities.

PMID:
25303369
PMCID:
PMC4220647
DOI:
10.1038/bcj.2014.69
[Indexed for MEDLINE]
Free PMC Article

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