Standardisation of operating procedures for the detection of minimal disease by QRT-PCR in children with neuroblastoma: Quality assurance on behalf of SIOPEN-R-NET

Virginie F. Viprey, Maria V. Corrias, Bertil Kagedal, Silvestre Oltra, Katrien Swerts, Aleš Vicha, Ruth Ladenstein, Susan A. Burchill

Research output: Contribution to journalArticlepeer-review

Abstract

The clinical utility of detecting minimal residual disease (MRD) in children with neuroblastoma (NB) by quantitative reverse transcriptase polymerase chain reaction (QRT-PCR) is not clear. This in part reflects the lack of uniform methodology for analysis and reporting. Reference laboratories across Europe have therefore established standard operating procedures (SOPs) for the detection of NB cells by QRT-PCR. Haemopoietic samples are collected into PAXgene™ blood RNA tubes, which stabilise mRNA for 48 h at room temperature and more than 6 months at -80 °C. Tyrosine hydroxylase (TH) was selected as the target for NB cell detection, expression is normalised to β2-microglobulin and reported using the ΔΔCt method. The sensitivity of QRT-PCR increased from 58% to 90% following the development of SOPs. A robust, transferable, objective method for the detection of NB cells by QRT-PCR has been defined to improve the power and consistency of studies on MRD in children with NB.

Original languageEnglish
Pages (from-to)341-350
Number of pages10
JournalEuropean Journal of Cancer
Volume43
Issue number2
DOIs
Publication statusPublished - Jan 2007

Keywords

  • β2-Microglobulin
  • Bone marrow
  • Neuroblastoma cells
  • PAXgene™ blood RNA kit
  • PAXgene™ blood RNA tubes
  • Peripheral blood
  • Peripheral blood stem cell harvests
  • Quantitative reverse transcriptase polymerase chain reaction (QRT-PCR)
  • Tyrosine hydroxylase mRNA

ASJC Scopus subject areas

  • Cancer Research
  • Hematology
  • Oncology

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