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|Title:||Cell-Free DNA in Pediatric Rhabdomyosarcoma: Potential and Challenges||Authors:||Tombolan, Lucia
|Keywords:||Cell-free DNA;Mutation detection;Pediatric solid tumors;Rhabdomyosarcoma;ddPCR||Mesh headings:||Cell-Free Nucleic Acids;DNA Mutational Analysis;Rhabdomyosarcoma||Secondary Mesh headings:||Child;Fluorometry;Humans;Mutation;Polymerase Chain Reaction||Issue Date:||2019||Journal:||Methods in molecular biology (Clifton, N.J.)||Abstract:||
Rhabdomyosarcoma is an aggressive solid tumor that may disseminate hematogenously giving metastasis which represents the most important prognostic factor. Chances of an effective cure in childhood cancer rely on the capacity to make an early and accurate diagnosis, detect metastatic disease or relapse, and predict the response to treatment.Liquid biopsy is a very promising blood test for cancer detection and noninvasive disease monitoring. This method has a great advantage to use blood and plasma, a more accessible biological material, quick and easy to obtain with minimal pain and risk for patients. In particular, circulating free DNA (cfDNA) represents a tumor biomarker detected in plasma that gives information on biology and genetic background of tumor.Moreover, cfDNA mutation detection could be a reliable method to monitor the efficacy of treatment and to discover novel targets for a personalized treatment in pediatric solid tumor. Here, we describe an optimized protocol to cfDNA isolation from small amount of plasma, as well as a method to assess the quantity and quality of cfDNA. Finally, we propose ddPCR as a reliable method to detect mutations at low frequency in cfDNA obtained from pediatric rhabdomyosarcoma samples.
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