title: 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.
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 degrees C. Tyrosine hydroxylase (TH) was selected as the target for NB cell detection, expression is normalised to beta2-microglobulin and reported using the DeltaDeltaCt 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.
authors.affiliation: Children's Cancer Research Laboratory, Cancer Research UK Clinical Centre, St James's University Hospital, Beckett Street, Leeds LS9 7TF, United Kingdom.
authors.forename: Virginie F
authors.initials: VF
authors.lastname: Viprey
authors.forename: Maria V
authors.initials: MV
authors.lastname: Corrias
authors.forename: Bertil
authors.initials: B
authors.lastname: Kagedal
authors.forename: Silvestre
authors.initials: S
authors.lastname: Oltra
authors.forename: Katrien
authors.initials: K
authors.lastname: Swerts
authors.forename: Ales
authors.initials: A
authors.lastname: Vicha
authors.forename: Ruth
authors.initials: R
authors.lastname: Ladenstein
authors.forename: Susan A
authors.initials: SA
authors.lastname: Burchill
mesh_terms: D045744:Cell Line, Tumor; D002648:Child; D002675:Child, Preschool; D006801:Humans; D007223:Infant; D018365:Neoplasm, Residual / Q000175:diagnosis*; D009447:Neuroblastoma / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D011786:Quality Control; D012334:RNA, Neoplasm / Q000032:analysis; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods / Q000592:standards
chemical_list: D012334:RNA, Neoplasm