title: Antibody detection of translocations in Ewing sarcoma.
abstract: The detection of chromosomal translocations has important implications in the diagnosis, prognosis and treatment of patients with cancer. Current approaches to translocation detection have significant shortcomings, including limited sensitivity and/or specificity, and difficulty in application to formalin-fixed paraffin-embedded (FFPE) clinical samples. We developed a new approach called antibody detection of translocations (ADOT) that avoids the shortcomings of current techniques. ADOT combines a transcriptional microarray-based approach with a novel antibody-based detection method. ADOT allows for the accurate and sensitive identification of translocations and provides exon-level information about the fusion transcript. ADOT can detect translocations in poor-quality unprocessed total ribonucleic acid (RNA). Furthermore, the technique is readily generalizable to detect any potential fusion transcript, including previously undescribed fusions. We demonstrate the feasibility of ADOT by examples in which both known and unknown Ewing sarcoma translocations are identified from cell lines, tumour xenografts and FFPE primary tumours. These results demonstrate that ADOT may be an effective approach for translocation analysis in clinical specimens with significant RNA degradation and may offer a novel diagnostic tool for translocation-based cancers.
authors.affiliation: The Department of Oncological Sciences, University of Utah School of Medicine, Salt Lake City, UT, USA.
authors.forename: Wen
authors.initials: W
authors.lastname: Luo
authors.forename: Brett
authors.initials: B
authors.lastname: Milash
authors.forename: Brian
authors.initials: B
authors.lastname: Dalley
authors.forename: Richard
authors.initials: R
authors.lastname: Smith
authors.forename: Holly
authors.initials: H
authors.lastname: Zhou
authors.forename: Natalie
authors.initials: N
authors.lastname: Dutrow
authors.forename: Bradley R
authors.initials: BR
authors.lastname: Cairns
authors.forename: Stephen L
authors.initials: SL
authors.lastname: Lessnick
mesh_terms: D000818:Animals; D000906:Antibodies; D050939:Gene Fusion; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D046228:Microarray Analysis / Q000379:methods; D057089:Pathology, Molecular / Q000379:methods*; D012512:Sarcoma, Ewing / Q000473:pathology*; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic; D014178:Translocation, Genetic
chemical_list: D000906:Antibodies