Analytical validation and implementation of a pan cancer next-generation sequencing panel, CANSeq TM Kids for molecular profiling of childhood malignancies.

Autor:	Schilter KF; Precision Medicine Laboratory, Department of Pathology, Medical College of Wisconsin, Milwaukee, WI, United States., Smith BA; Precision Medicine Laboratory, Department of Pathology, Medical College of Wisconsin, Milwaukee, WI, United States., Nie Q; Precision Medicine Laboratory, Department of Pathology, Medical College of Wisconsin, Milwaukee, WI, United States., Stoll K; Precision Medicine Laboratory, Department of Pathology, Medical College of Wisconsin, Milwaukee, WI, United States., Felix JC; Precision Medicine Laboratory, Department of Pathology, Medical College of Wisconsin, Milwaukee, WI, United States.; Pathology and Laboratory Medicine, Medical College of Wisconsin, Milwaukee, WI, United States., Jarzembowski JA; Pathology and Laboratory Medicine, Medical College of Wisconsin, Milwaukee, WI, United States., Reddi HV; Precision Medicine Laboratory, Department of Pathology, Medical College of Wisconsin, Milwaukee, WI, United States.
Jazyk:	angličtina
Zdroj:	Frontiers in genetics [Front Genet] 2023 Feb 09; Vol. 14, pp. 1067457. Date of Electronic Publication: 2023 Feb 09 (Print Publication: 2023).
DOI:	10.3389/fgene.2023.1067457
Abstrakt:	Next-Generation Sequencing (NGS) allows rapid analysis of multiple genes for the detection of clinically actionable variants. This study reports the analytical validation of a targeted pan cancer NGS panel CANSeq TM Kids for molecular profiling of childhood malignancies. Analytical validation included DNA and RNA extracted from de-identified clinical specimens including formalin fixed paraffin embedded (FFPE) tissue, bone marrow and whole blood as well as commercially available reference materials. The DNA component of the panel evaluates 130 genes for the detection of single nucleotide variants (SNVs), Insertion and Deletions (INDELs), and 91 genes for fusion variants associated with childhood malignancies. Conditions were optimized to use as low as 20% neoplastic content with 5 ng of nucleic acid input. Evaluation of the data determined greater than 99% accuracy, sensitivity, repeatability, and reproducibility. The limit of detection was established to be 5% allele fraction for SNVs and INDELs, 5 copies for gene amplifications and 1,100 reads for gene fusions. Assay efficiency was improved by automation of library preparation. In conclusion, the CANSeq TM Kids allows for the comprehensive molecular profiling of childhood malignancies from different specimen sources with high quality and fast turnaround time. Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2023 Schilter, Smith, Nie, Stoll, Felix, Jarzembowski and Reddi.)
Databáze:	MEDLINE
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