Molecular Diagnosis of Ewing TumorsImproved Detection of EWS‐FLI‐1 and EWS‐ERG Chimeric Transcripts and Rapid Determination of Exon Combinations
作者:
Verena,
Meier Thomas,
Kühne Gernot,
Jundt Fred,
期刊:
Diagnostic Molecular Pathology
(OVID Available online 1998)
卷期:
Volume 7,
issue 1
页码: 29-35
ISSN:1052-9551
年代: 1998
出版商: OVID
关键词: Ewing tumors;1( 11;22)(q24;q12);t(21;22)(q21:q12);EWS gene;FL1;1 gene;ERG gene
数据来源: OVID
摘要:
Most Ewing tumors (ET), including Ewing sarcomas, peripheral primitive neuroectodermal tumors (PNET), and Askin's tumors, can be defined according to the specific chromosomal translocations t(11; 22)(q24;q12) (EWS-FLI-1) or t(21;22)(q21; q12) (EWS-ERG).Detection of the chimeric RNA transcripts by reverse transcriptase-polymerase chain reaction (RT-PCR) has greatly facilitated the diagnosis of ET. Because of variable chromosomal breakpoint locations, however, the EWS gene fusions with FLI-I and ERG genes are highly heterogenous, resulting in different sizes of the amplification products. To improve the diagnostic usefulness of the RT-PCR assay, we have developed an assay to detect chimeric mRNA transcripts by nested RT-PCR, followed by digestion of the PCR fragments with three different restriction endonucleases. This allows confirmation of the specificity of the PCR product and provides a rapid method to determine the combination of exons present in a transcript. In the 12 Ewing tumors tested, five different exon combinations were detected. In nine repeat biopsies of four patients, the case-specific translocation remained unchanged. One additional central PNET had no ET-specific translocation. In conclusion, the suggested combination of RT-PCR and restriction analysis of the PCR products allows a rapid and specific determination of ET-specific translocations.
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