A method for accurate detection of translocation junctions in Ewing family of tumors.
The Ewing family of tumors (ET) generally contain translocations involving the EWS gene and the FLI or ERG genes. Identification of the translocation confirms the diagnosis of ET. Currently, diagnosis of the translocation is made by several methods. In general, these methods require different primer sets for amplifying different translocations and subsequent efforts to identify the amplified product. The need to employ different sets of primers to amplify different translocation junctions presents some limitations. We have developed a method based on PCR with consensus primers followed by direct automated sequencing of the amplified product. With this method we have correctly determined known as well as unknown ET-associated EWS-FLI and EWS-ERG translocations in appropriate specimens. Use of our consensus primers eliminates the need for separate PCRs to amplify EWS-FLI and EWS-ERG translocation junctions, and because direct sequencing is used for confirming the identity of the amplification product, the accuracy of detection becomes 100%. The method might also accurately diagnose ET-associated translocations other than EWS-FLI and EWS-ERG translocations.
Morishita
T
T
Mayo Clinic, 3-15, Medical Sciences Building, Rochester, MN 55905, USA.
Bolander
M E
ME
Zhang
K
K
Tamai
S
S
Mii
Y
Y
Sarkar
G
G
D000595:Amino Acid Sequence; D001483:Base Sequence; D027682:Cation Transport Proteins; D019457:Chromosome Breakage / Q000235:genetics*; D016384:Consensus Sequence / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D015514:Oncogene Proteins, Fusion / Q000235:genetics; D015221:Potassium Channels / Q000235:genetics; D024642:Potassium Channels, Voltage-Gated; D051789:Proto-Oncogene Protein c-fli-1; D011518:Proto-Oncogene Proteins; D034802:RNA-Binding Protein EWS; D016364:Reading Frames / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012261:Ribonucleoproteins / Q000235:genetics; D012512:Sarcoma, Ewing / Q000175:diagnosis* / Q000235:genetics*; D017422:Sequence Analysis, DNA; D015534:Trans-Activators / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D000071230:Transcriptional Regulator ERG; D014178:Translocation, Genetic / Q000235:genetics*; D014407:Tumor Cells, Cultured
D027682:Cation Transport Proteins; D017931:DNA Primers; D004268:DNA-Binding Proteins; C072312:ERG protein, human; D000072237:ERG1 Potassium Channel; C106031:EWS-FLI fusion protein; D051638:Ether-A-Go-Go Potassium Channels; D034441:Heterogeneous-Nuclear Ribonucleoproteins; C092509:KCNH6 protein, human; D015514:Oncogene Proteins, Fusion; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D051789:Proto-Oncogene Protein c-fli-1; D011518:Proto-Oncogene Proteins; D034802:RNA-Binding Protein EWS; D012261:Ribonucleoproteins; D015534:Trans-Activators; D014157:Transcription Factors; D000071230:Transcriptional Regulator ERG