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A highly sensitive and specific qPCR assay for quantification of the biomarker SOX11 in mantle cell lymphoma
Author(s) -
Hamborg Kristiane Hornung,
Bentzen Hans Herluf Nørregaard,
Grubach Lykke,
Hokland Peter,
Nyvold Charlotte Guldborg
Publication year - 2012
Publication title -
european journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.904
H-Index - 84
eISSN - 1600-0609
pISSN - 0902-4441
DOI - 10.1111/j.1600-0609.2012.01837.x
Subject(s) - mantle cell lymphoma , minimal residual disease , microbiology and biotechnology , biomarker , bone marrow , cyclin d1 , peripheral blood mononuclear cell , real time polymerase chain reaction , cancer research , lymphoma , chemistry , biology , immunology , cell , in vitro , cell cycle , gene , biochemistry
Objectives Mantle cell lymphoma ( MCL ) is one of the most heterogeneous lymphoid neoplasms with a variable course of disease. Although t(11;14)(q13;q32) is the hallmark of MCL resulting in cyclin D1 ( CCND 1 ) overexpression in 90% of patients, this is difficult to validate by immunohistochemistry. We hypothesised that SOX 11 could be a robust molecular biomarker for MCL . Methods We have developed very sensitive and specific RT‐ qPCR assay employing a poly‐A specific RT primer to circumvent contamination from gDNA caused by the intron‐less nature of SOX11 . Results We found a significant difference between the expression levels of SOX 11 in patients with MCL at diagnosis ( n = 21) and in healthy donors ( n = 18) (blood: P < 0.0001; marrow: P = 0.0001). SOX 11 expression of very low levels close to the assay sensitivity was detected in only 2 of 18 healthy donors, while low levels of CCND 1 expression was observed in all blood and 12 of 13 marrow samples within the defined detection limit of Cq = 40. In spiking experiments of the GRANTA ‐519 MCL cell line into mononuclear cells from normal donor, the sensitivity of the SOX 11 assay was found to be 2 × 10 −4 , while the sensitivity of the CCND 1 assay was estimated to 2 × 10 −3 because of the normal background expression. In longitudinal sampling from patients with MCL the minimal residual disease ( MRD ) values based on the SOX 11 expression mirrored the clinical disease development. Conclusion This SOX11 RT‐qPCR assay could be a useful tool for MRD monitoring in patients with MCL.