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

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 sequenci...

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Autores principales:	Hitomi Ueno-Yokohata, Hajime Okita, Keiko Nakasato, Chikako Kiyotani, Motohiro Kato, Kimikazu Matsumoto, Nobutaka Kiyokawa, Atsuko Nakazawa, Takako Yoshioka
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Ewing sarcoma Multiplex RT–PCR Genetic diagnosis Fusion gene EWSR1 Transcription factor Pathology RB1-214
Acceso en línea:	https://doaj.org/article/15b950b301c143b0be4a11442dd56f90
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spelling	oai:doaj.org-article:15b950b301c143b0be4a11442dd56f902021-11-14T12:37:44ZEstablishment of multiplex RT-PCR to detect fusion genes for the diagnosis of Ewing sarcoma10.1186/s13000-021-01164-61746-1596https://doaj.org/article/15b950b301c143b0be4a11442dd56f902021-11-01T00:00:00Zhttps://doi.org/10.1186/s13000-021-01164-6https://doaj.org/toc/1746-1596Abstract 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.Hitomi Ueno-YokohataHajime OkitaKeiko NakasatoChikako KiyotaniMotohiro KatoKimikazu MatsumotoNobutaka KiyokawaAtsuko NakazawaTakako YoshiokaBMCarticleEwing sarcomaMultiplex RT–PCRGenetic diagnosisFusion geneEWSR1Transcription factorPathologyRB1-214ENDiagnostic Pathology, Vol 16, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Ewing sarcoma Multiplex RT–PCR Genetic diagnosis Fusion gene EWSR1 Transcription factor Pathology RB1-214
spellingShingle	Ewing sarcoma Multiplex RT–PCR Genetic diagnosis Fusion gene EWSR1 Transcription factor Pathology RB1-214 Hitomi Ueno-Yokohata Hajime Okita Keiko Nakasato Chikako Kiyotani Motohiro Kato Kimikazu Matsumoto Nobutaka Kiyokawa Atsuko Nakazawa Takako Yoshioka Establishment of multiplex RT-PCR to detect fusion genes for the diagnosis of Ewing sarcoma
description	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.
format	article
author	Hitomi Ueno-Yokohata Hajime Okita Keiko Nakasato Chikako Kiyotani Motohiro Kato Kimikazu Matsumoto Nobutaka Kiyokawa Atsuko Nakazawa Takako Yoshioka
author_facet	Hitomi Ueno-Yokohata Hajime Okita Keiko Nakasato Chikako Kiyotani Motohiro Kato Kimikazu Matsumoto Nobutaka Kiyokawa Atsuko Nakazawa Takako Yoshioka
author_sort	Hitomi Ueno-Yokohata
title	Establishment of multiplex RT-PCR to detect fusion genes for the diagnosis of Ewing sarcoma
title_short	Establishment of multiplex RT-PCR to detect fusion genes for the diagnosis of Ewing sarcoma
title_full	Establishment of multiplex RT-PCR to detect fusion genes for the diagnosis of Ewing sarcoma
title_fullStr	Establishment of multiplex RT-PCR to detect fusion genes for the diagnosis of Ewing sarcoma
title_full_unstemmed	Establishment of multiplex RT-PCR to detect fusion genes for the diagnosis of Ewing sarcoma
title_sort	establishment of multiplex rt-pcr to detect fusion genes for the diagnosis of ewing sarcoma
publisher	BMC
publishDate	2021
url	https://doaj.org/article/15b950b301c143b0be4a11442dd56f90
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Establishment of multiplex RT-PCR to detect fusion genes for the diagnosis of Ewing sarcoma

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