Application of mediator probe PCR chemistry in multiplex target assays for monitoring MRD of high-risk neuroblastoma

Patients with high-risk neuroblastoma show in about 85% a bone marrow infiltration. Even though most of them achieve remission, minimal residual disease (MRD) frequently persists on submicroscopic level and causes a relapse. Our aim was to detect MRD cells in bone marrow aspirates taken during treatment, by employing a novel sensitive multiplex PCR chemistry, the mediator probe PCR (MP-PCR), which consists of a target-specific, unlabeled mediator probe and sequence-universal reporter conjugated with a fluorescent dye. Previously, the MP-PCR was successfully applied by Kipf et al. to determine MRD levels of acute lymphoblastic leukemia. Comparably to leukemia, high-risk neuroblastoma harbor multiple genetic alterations, which in addition appear on extrachromosomal DNA, including MYCN breakpoints, TERT rearrangements and ALK mutations. Our patient cohort comprises 18 patients with MYCN amplified neuroblastoma with multiple MYCN breakpoints and other mutations. To capture this tumor heterogeneity, we combined up to four MRD markers in one multiplex assay. Sensitive quantification of MRD benefits for the reconstruction of the clonal biology and personalization of neuroblastoma therapy.

Publication History

Article published online:
17 May 2022