title: A Rapid and Sensitive Next-Generation Sequencing Method to Detect RB1 Mutations Improves Care for Retinoblastoma Patients and Their Families.
abstract: Retinoblastoma is a childhood eye malignancy that can lead to the loss of vision, eye(s), and sometimes life. The tumors are initiated by inactivating mutations in both alleles of the tumor-suppressor gene, RB1, or, rarely, by MYCN amplification. Timely identification of a germline RB1 mutation in blood samples or either somatic RB1 mutation or MYCN amplification in tumors is important for effective care and management of retinoblastoma patients and their families. However, current procedures to thoroughly test RB1 mutations are complicated and lengthy. Herein, we report a next-generation sequencing-based method capable of detecting point mutations, small indels, and large deletions or duplications across the entire RB1 gene and amplification of MYCN gene on a single platform. From DNA extraction to clinical interpretation requires only 3 days, enabling early molecular diagnosis of retinoblastoma and optimal treatment outcomes. This method can also detect low-level mosaic mutations in blood samples that can be missed by routine Sanger sequencing. In addition, it can differentiate between RB1 mutation- and MYCN amplification-driven retinoblastomas. This rapid, comprehensive, and sensitive method for detecting RB1 mutations and MYCN amplification can readily identify RB1 mutation carriers and thus improve the management and genetic counseling for retinoblastoma patients and their families.
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital Los Angeles, Los Angeles, California; Department of Pathology, USC Roski Eye Institute, University of Southern California, Los Angeles, California. Electronic address: lauli@chla.usc.edu.
authors.forename: Wenhui L
authors.initials: WL
authors.lastname: Li
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital Los Angeles, Los Angeles, California; Department of Pathology, USC Roski Eye Institute, University of Southern California, Los Angeles, California.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Buckley
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital Los Angeles, Los Angeles, California; Department of Pathology, USC Roski Eye Institute, University of Southern California, Los Angeles, California; Department of Pediatrics, USC Roski Eye Institute, University of Southern California, Los Angeles, California.
authors.forename: Pedro A
authors.initials: PA
authors.lastname: Sanchez-Lara
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital Los Angeles, Los Angeles, California.
authors.forename: Dennis T
authors.initials: DT
authors.lastname: Maglinte
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital Los Angeles, Los Angeles, California.
authors.forename: Lucy
authors.initials: L
authors.lastname: Viduetsky
authors.affiliation: Department of Pediatrics, USC Roski Eye Institute, University of Southern California, Los Angeles, California; Spatial Sciences Institute, Dornsife College of Letters, Arts and Sciences, University of Southern California, Los Angeles, California.
authors.forename: Tatiana V
authors.initials: TV
authors.lastname: Tatarinova
authors.affiliation: Vision Center, Children's Hospital Los Angeles, Los Angeles, California.
authors.forename: Jennifer G
authors.initials: JG
authors.lastname: Aparicio
authors.affiliation: Vision Center, Children's Hospital Los Angeles, Los Angeles, California; Department of Opthalmology, USC Roski Eye Institute, University of Southern California, Los Angeles, California.
authors.forename: Jonathan W
authors.initials: JW
authors.lastname: Kim
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital Los Angeles, Los Angeles, California.
authors.forename: Margaret
authors.initials: M
authors.lastname: Au
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital Los Angeles, Los Angeles, California.
authors.forename: Dejerianne
authors.initials: D
authors.lastname: Ostrow
authors.affiliation: Vision Center, Children's Hospital Los Angeles, Los Angeles, California; Department of Opthalmology, USC Roski Eye Institute, University of Southern California, Los Angeles, California.
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Lee
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital Los Angeles, Los Angeles, California; Department of Pathology, USC Roski Eye Institute, University of Southern California, Los Angeles, California.
authors.forename: Maurice
authors.initials: M
authors.lastname: O'Gorman
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital Los Angeles, Los Angeles, California; Department of Pathology, USC Roski Eye Institute, University of Southern California, Los Angeles, California.
authors.forename: Alexander
authors.initials: A
authors.lastname: Judkins
authors.affiliation: Vision Center, Children's Hospital Los Angeles, Los Angeles, California; Department of Opthalmology, USC Roski Eye Institute, University of Southern California, Los Angeles, California; Division of Ophthalmology and Department of Surgery, and Saban Research Institute, Children's Hospital Los Angeles, Los Angeles, California; Department of Biochemistry & Molecular Biology, USC Roski Eye Institute, University of Southern California, Los Angeles, California; Norris Comprehensive Cancer Center, USC Keck School of Medicine, University of Southern California, Los Angeles, California.
authors.forename: David
authors.initials: D
authors.lastname: Cobrinik
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital Los Angeles, Los Angeles, California; Department of Pathology, USC Roski Eye Institute, University of Southern California, Los Angeles, California. Electronic address: ttriche@chla.usc.edu.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Triche
mesh_terms: D000483:Alleles; D056915:DNA Copy Number Variations; D004252:DNA Mutational Analysis / Q000379:methods; D005091:Exons; D017353:Gene Deletion; D020440:Gene Duplication; D016161:Genes, Retinoblastoma; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D009030:Mosaicism; D009154:Mutation; D011401:Promoter Regions, Genetic; D012175:Retinoblastoma / Q000175:diagnosis* / Q000235:genetics*