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Quantitative PCR: an alternative approach to detect common copy number alterations in multiple myeloma

AuthorsChillón, M. del Carmen; Jiménez, Cristina ; García-Sanz, Ramón; Alcoceba, Miguel; Prieto-Conde, Isabel; García-Alvarez, María; Antón, Alicia; Maldonado, R.; Hernández-Ruano, M.; González, Marcos ; Gutiérrez, Norma Carmen; Sarasquete, María Eugenia
KeywordsInterphase fluorescence in situ hybridization
Copy number variation
Quantitative PCR
Multiple myeloma
Issue Date2017
PublisherSpringer Nature
CitationAnnals of Hematology 96(10): 1699-1705 (2017)
AbstractChromosome 1q gains and 13q deletions are common cytogenetic aberrations in multiple myeloma (MM) that confer a poor prognosis. There are several techniques for the targeted study of these alterations, but interphase fluorescence in situ hybridization (FISH) is the current gold standard. The aim of the present study was to validate quantitative PCR (qPCR) as an alternative to FISH studies in CD138+-enriched plasma cells (PCs) from MM patients at diagnosis. We analyzed 1q gains and 13q deletions by qPCR in 57 and 60 MM patients, respectively. qPCR applicability was 84 and 88% for 1q and 13q, respectively. The qPCR and FISH methods had a sensitivity and specificity of 88 and 71% for 1q gains, and 79 and 100% for 13q deletions. A second qPCR assay for each region was carried out to confirm the previous results. Paired qPCR (two assays) and FISH results were available from 53 MM patients: 26 for 1q amplification and 27 for 13q deletion. qPCR assays gave concordant results (qPCR-consistent) in 20 of the 26 (77%) 1q gains and 25 of the 27 (93%) 13q deletions. Considering only the consistent data, the overall concordance among qPCR and FISH was 85 and 100% for 1q gains and 13q deletions, respectively. Our results show a substantial agreement between qPCR and the gold standard FISH technique, indicating the potential of qPCR as an alternative approach, particularly when the starting material is too scarce or cells are too damaged to obtain accurate results from FISH studies.
Identifiersdoi: 10.1007/s00277-017-3083-x
e-issn: 1432-0584
issn: 0939-5555
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