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Stability of PCR Targets for Monitoring Minimal Residual Disease in Neuroblastoma

Stutterheim, Janine; Zappeij-Kannegieter, Lily; Øra, Ingrid LU ; van Sluis, Peter G.; Bras, Johannes; den Ouden, Emmy; Versteeg, Rogier; Caron, Huib N.; van der Schoot, C. Ellen and Tytgat, Godelieve A. M. (2012) In The Journal Of Molecular Diagnostics 14(2). p.168-175
Abstract
In neuroblastoma (NB) patients, minimal residual disease (MRD) can be detected by real-time quantitative PCR (qPCR) using NB-specific target genes, such as PHOX2B and TH. However, it is unknown whether the mRNA levels of these targets vary either during treatment or at relapse. If marker genes are not stably expressed, estimation of MRD levels in bone marrow (BM) or peripheral blood will be hampered. We studied the stability of a panel of qPCR markers in primary tumors at diagnosis compared with i) paired metastasis (n = 7), ii) treated (n = 10), and iii) relapse (n = 6) tumors. We also compared relative expression of the targets in iv) primary tumors and BM at diagnosis (n = 17), v) BM and peripheral blood at diagnosis (n = 20), vi) BM at... (More)
In neuroblastoma (NB) patients, minimal residual disease (MRD) can be detected by real-time quantitative PCR (qPCR) using NB-specific target genes, such as PHOX2B and TH. However, it is unknown whether the mRNA levels of these targets vary either during treatment or at relapse. If marker genes are not stably expressed, estimation of MRD levels in bone marrow (BM) or peripheral blood will be hampered. We studied the stability of a panel of qPCR markers in primary tumors at diagnosis compared with i) paired metastasis (n = 7), ii) treated (n = 10), and iii) relapse (n = 6) tumors. We also compared relative expression of the targets in iv) primary tumors and BM at diagnosis (n = 17), v) BM and peripheral blood at diagnosis (n = 20), vi) BM at diagnosis and during treatment (n = 26), and vii) BM from different puncture sides (n = 110). Especially at diagnosis, PCR target expression is quite stable. Accurate quantification is possible when expression level can be related to the primary tumor; however, PCR target expression can alter on treatment and at relapse. If the median value of relative expression of a panel of PCR targets is used, most variations due to treatment and outgrowth of subclones level out, allowing for reliable application and quantification of MRD-PCR targets in NB patients. (J Mol Diagn 2012, 14:168-175; DOI: 10.1016/j.jmoldx.2011.12.002) (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
The Journal Of Molecular Diagnostics
volume
14
issue
2
pages
168 - 175
publisher
Elsevier Inc.
external identifiers
  • wos:000301007700009
  • scopus:84857284549
ISSN
1525-1578
DOI
10.1016/j.jmoldx.2011.12.002
language
English
LU publication?
yes
id
8ac0ee5c-85f2-4346-9365-eeed0739f8b8 (old id 2379345)
date added to LUP
2012-04-02 09:22:28
date last changed
2017-06-25 03:59:41
@article{8ac0ee5c-85f2-4346-9365-eeed0739f8b8,
  abstract     = {In neuroblastoma (NB) patients, minimal residual disease (MRD) can be detected by real-time quantitative PCR (qPCR) using NB-specific target genes, such as PHOX2B and TH. However, it is unknown whether the mRNA levels of these targets vary either during treatment or at relapse. If marker genes are not stably expressed, estimation of MRD levels in bone marrow (BM) or peripheral blood will be hampered. We studied the stability of a panel of qPCR markers in primary tumors at diagnosis compared with i) paired metastasis (n = 7), ii) treated (n = 10), and iii) relapse (n = 6) tumors. We also compared relative expression of the targets in iv) primary tumors and BM at diagnosis (n = 17), v) BM and peripheral blood at diagnosis (n = 20), vi) BM at diagnosis and during treatment (n = 26), and vii) BM from different puncture sides (n = 110). Especially at diagnosis, PCR target expression is quite stable. Accurate quantification is possible when expression level can be related to the primary tumor; however, PCR target expression can alter on treatment and at relapse. If the median value of relative expression of a panel of PCR targets is used, most variations due to treatment and outgrowth of subclones level out, allowing for reliable application and quantification of MRD-PCR targets in NB patients. (J Mol Diagn 2012, 14:168-175; DOI: 10.1016/j.jmoldx.2011.12.002)},
  author       = {Stutterheim, Janine and Zappeij-Kannegieter, Lily and Øra, Ingrid and van Sluis, Peter G. and Bras, Johannes and den Ouden, Emmy and Versteeg, Rogier and Caron, Huib N. and van der Schoot, C. Ellen and Tytgat, Godelieve A. M.},
  issn         = {1525-1578},
  language     = {eng},
  number       = {2},
  pages        = {168--175},
  publisher    = {Elsevier Inc.},
  series       = {The Journal Of Molecular Diagnostics},
  title        = {Stability of PCR Targets for Monitoring Minimal Residual Disease in Neuroblastoma},
  url          = {http://dx.doi.org/10.1016/j.jmoldx.2011.12.002},
  volume       = {14},
  year         = {2012},
}