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Open Access 01-12-2021 | Fluorescence in Situ Hybridization | Research

Establishment of multiplex RT-PCR to detect fusion genes for the diagnosis of Ewing sarcoma

Authors: Hitomi Ueno-Yokohata, Hajime Okita, Keiko Nakasato, Chikako Kiyotani, Motohiro Kato, Kimikazu Matsumoto, Nobutaka Kiyokawa, Atsuko Nakazawa, Takako Yoshioka

Published in: Diagnostic Pathology | Issue 1/2021

Abstract

Background

Detection of the tumor-specific EWSR1/FUS-ETS fusion gene is essential to diagnose Ewing sarcoma. Reverse transcription–polymerase chain reaction (RT–PCR) and fluorescence in situ hybridization are commonly used to detect the fusion gene, and assays using next-generation sequencing have recently been reported. However, at least 28 fusion transcript variants have been reported, making rapid and accurate detection difficult.

Methods

We constructed two sets of multiplex PCR assays and evaluated their utility using cell lines and clinical samples.

Results

EWSR1/FUS-ETS was detected in five of six tumors by the first set, and in all six tumors by the second set. The fusion gene detected only by the latter was EWSR1-ERG, which completely lacked exon 7 of EWSR1. The fusion had a short N-terminal region of EWSR1 and showed pathologically atypical features.

Conclusions

We developed multiplex RT–PCR assays to detect EWSR1-ETS and FUS-ETS simultaneously. These assays will aid the rapid and accurate diagnosis of Ewing sarcoma. In addition, variants of EWSR1/FUS-ETS with a short N-terminal region that may have been previously missed can be easily detected.

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Title: Establishment of multiplex RT-PCR to detect fusion genes for the diagnosis of Ewing sarcoma
Authors: Hitomi Ueno-Yokohata
Hajime Okita
Keiko Nakasato
Chikako Kiyotani
Motohiro Kato
Kimikazu Matsumoto
Nobutaka Kiyokawa
Atsuko Nakazawa
Takako Yoshioka
Publication date: 01-12-2021
Publisher: BioMed Central
Keywords: Fluorescence in Situ Hybridization
Sarcoma
Published in: Diagnostic Pathology / Issue 1/2021
Electronic ISSN: 1746-1596
DOI: https://doi.org/10.1186/s13000-021-01164-6

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Establishment of multiplex RT-PCR to detect fusion genes for the diagnosis of Ewing sarcoma

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

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