TY - JOUR
T1 - Specific and sensitive detection of neuroblastoma mRNA markers by multiplex RT-qPCR
AU - van Zogchel, Lieke M. J.
AU - Zappeij-Kannegieter, Lily
AU - Javadi, Ahmad
AU - Lugtigheid, Marjolein
AU - Gelineau, Nina U.
AU - Lak, Nathalie S. M.
AU - Zwijnenburg, Danny A.
AU - Koster, Jan
AU - Stutterheim, Janine
AU - Ellen van der Schoot, C.
AU - Tytgat, Godelieve A. M.
N1 - Funding Information: Funding: This research was funded by Liquidhope, a TranScan-2 project by Koningin Wilhelmina Fund, KWF Kankerbestrijding TRANSCAN 8352/TRS-2018-00000715 (L.M.J.v.Z, N.U.G, J.K.) and Foundation AMeesing Mees. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/1/1
Y1 - 2021/1/1
N2 - mRNA RT-qPCR is shown to be a very sensitive technique to detect minimal residual disease (MRD) in patients with neuroblastoma. Multiple mRNA markers are known to detect heterogeneous neuroblastoma cells in bone marrow (BM) or blood from patients. However, the limited volumes of BM and blood available can hamper the detection of multiple markers. To make optimal use of these samples, we developed a multiplex RT-qPCR for the detection of MRD in neuroblastoma. GUSB and PHOX2B were tested as single markers. The adrenergic markers TH, GAP43, CHRNA3 and DBH and mesenchymal markers POSTN, PRRX1 and FMO3 were tested in multiplex. Using control blood and BM, we established new thresholds for positivity. Comparison of multiplex and singleplex RT-qPCR results from 21 blood and 24 BM samples from neuroblastoma patients demonstrated a comparable sensitivity. With this multiplex RT-qPCR, we are able to test seven different neuroblastoma mRNA markers, which overcomes tumor heterogeneity and improves sensitivity of MRD detection, even in those samples of low RNA quantity. With resources and time being saved, reduction in sample volume and consumables can assist in the introduction of MRD by RT-qPCR into clinical practice.
AB - mRNA RT-qPCR is shown to be a very sensitive technique to detect minimal residual disease (MRD) in patients with neuroblastoma. Multiple mRNA markers are known to detect heterogeneous neuroblastoma cells in bone marrow (BM) or blood from patients. However, the limited volumes of BM and blood available can hamper the detection of multiple markers. To make optimal use of these samples, we developed a multiplex RT-qPCR for the detection of MRD in neuroblastoma. GUSB and PHOX2B were tested as single markers. The adrenergic markers TH, GAP43, CHRNA3 and DBH and mesenchymal markers POSTN, PRRX1 and FMO3 were tested in multiplex. Using control blood and BM, we established new thresholds for positivity. Comparison of multiplex and singleplex RT-qPCR results from 21 blood and 24 BM samples from neuroblastoma patients demonstrated a comparable sensitivity. With this multiplex RT-qPCR, we are able to test seven different neuroblastoma mRNA markers, which overcomes tumor heterogeneity and improves sensitivity of MRD detection, even in those samples of low RNA quantity. With resources and time being saved, reduction in sample volume and consumables can assist in the introduction of MRD by RT-qPCR into clinical practice.
KW - Metastasis
KW - Minimal residual disease
KW - Neuroblastoma
KW - RT-qPCR
UR - http://www.scopus.com/inward/record.url?scp=85099952563&partnerID=8YFLogxK
U2 - https://doi.org/10.3390/cancers13010150
DO - https://doi.org/10.3390/cancers13010150
M3 - Article
C2 - 33466359
SN - 2072-6694
VL - 13
SP - 1
EP - 12
JO - Cancers
JF - Cancers
IS - 1
M1 - 150
ER -