title: Detection of tumor ALK status in neuroblastoma patients using peripheral blood.
abstract: New protocols based on ALK-targeted therapy by crizotinib or other ALK-targeting molecules have opened for the treatment of patients with neuroblastoma (NB) if their tumors showed mutation and/or amplification of the ALK gene. However, tumor samples are not always available for analysis of ALK mutational status in particular at relapse. Here, we evaluated the ALK mutational status of NB samples by analysis of circulating DNA, using the droplet digital PCR (ddPCR) system. ddPCR assays was developed for the detection of ALK mutations at F1174 and R1275 hotspots found in NB tumors and was applied for the analysis of circulating DNA obtained from 200 μL of serum or plasma samples collected from 114 patients with NB. The mutations F1174L (exon 23 position 3520, T>C and position 3522, C>A) and the mutation R1275Q (exon 25 position 3824, G>A) were detected in circulating DNA. The sensitivity of our test was 100%, 85%, and 92%, respectively, and the specificity was 100%, 91%, and 98%, respectively. In conclusion, the assay that we have developed offers a reliable, noninvasive blood test to assess ALK mutational status at F1174 and R1275 hotspots and should help clinicians to identify patients showing an ALK mutation in particular when no tumor tissue is available.
authors: Centre Léon-Bérard, Laboratoire de Recherche Translationnelle, 28 rue Laennec, 69373, Lyon Cedex 08, France.
authors: Valérie
authors: V
authors: Combaret
authors: Isabelle
authors: I
authors: Iacono
authors: Angela
authors: A
authors: Bellini
authors: Stéphanie
authors: S
authors: Bréjon
authors: Virginie
authors: V
authors: Bernard
authors: Aurélien
authors: A
authors: Marabelle
authors: Carole
authors: C
authors: Coze
authors: Gaelle
authors: G
authors: Pierron
authors: Eve
authors: E
authors: Lapouble
authors: Gudrun
authors: G
authors: Schleiermacher
authors: Jean Yves
authors: JY
authors: Blay
mesh_terms: D016022:Case-Control Studies; D002648:Child; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000235:genetics; D005091:Exons; D006801:Humans; D009154:Mutation / Q000235:genetics*; D009447:Neuroblastoma / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics*
chemical_list: D004273:DNA, Neoplasm; D020794:Receptor Protein-Tyrosine Kinases
keywords: ALK mutation; cell-free DNA; ddPCR; neuroblastoma
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