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Updated: Apr 17, 2026
Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids
Published on: June 8, 2019
Detection of tumor ALK status in neuroblastoma patients using peripheral blood
Valérie Combaret1, Isabelle Iacono, Angela Bellini
1Centre Léon-Bérard, Laboratoire de Recherche Translationnelle, 28 rue Laennec, 69373, Lyon Cedex 08, France.
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.
Insights
A new noninvasive blood test using droplet digital PCR (ddPCR) accurately detects ALK gene mutations in neuroblastoma (NB) patients. This method is crucial for identifying patients eligible for ALK-targeted therapy, especially when tumor tissue is unavailable.
Area of Science:
- Oncology
- Molecular Diagnostics
- Genetics
Background:
- Anaplastic Lymphoma Kinase (ALK) targeted therapies, like crizotinib, are emerging treatments for neuroblastoma (NB).
- Assessing ALK mutational status is critical for patient selection, but tumor tissue is often inaccessible, particularly during relapse.
Purpose of the Study:
- To develop and validate a noninvasive method for detecting ALK gene mutations in NB using circulating DNA.
- To evaluate the reliability of droplet digital PCR (ddPCR) for identifying specific ALK hotspots (F1174 and R1275) in NB patients' blood.
Main Methods:
- Developed ddPCR assays to detect ALK mutations at F1174 and R1275 hotspots.
- Analyzed circulating DNA from 200 μL of serum or plasma samples from 114 NB patients.
- Validated assay sensitivity and specificity for detecting F1174L and R1275Q mutations.
Main Results:
- Successfully detected ALK mutations F1174L and R1275Q in the circulating DNA of NB patients.
- Achieved high sensitivity (100%, 85%, 92%) and specificity (100%, 91%, 98%) for the developed ddPCR assays.
- Demonstrated the feasibility of using small blood volumes (200 μL) for mutation detection.
Conclusions:
- The developed ddPCR assay provides a reliable, noninvasive method to assess ALK mutational status in NB patients.
- This blood test can aid clinicians in identifying patients with actionable ALK mutations, especially when tumor tissue is unavailable.
- Facilitates timely initiation of ALK-targeted therapies for neuroblastoma treatment.
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