Download the poster "Verification of an Ion AmpliSeq™ RNA Fusion Lung Cancer Research Panel, workflow, and analysis solution: an OncoNetwork collaborative research study" on Slideshare: [ Ссылка ]
Fusion transcripts resulting from translocation events in the oncogenic driver genes ALK, RET, ROS1, and NTRK1 play an important role in lung adenocarcinoma. There is a need to detect these fusion transcripts with up to date technologies as they may serve as viable therapeutic targets. We have utilized a targeted sequencing approach and developed an Ion AmpliSeq™ RNA Lung Fusion panel, a workflow, and an Ion Reporter™ analysis solution to detect these known fusion events. The panel detects transcripts from 37 ALK, 9 RET, 15 ROS1, and 11 NTRK fusion variants along with 5 housekeeping genes to serve as internal controls. The workflow is FFPE compatible requiring an input of only 10 ng of total RNA with the capacity to multiplex up to 16 libraries on a single Ion 318™ chip. The panel was initially validated using 10ng of total RNA from a cocktail of 3 cell lines containing known lung cancer fusions (H2228 – EML4-ALK variant 3a and 3b, HCC78 – SLC34A2-ROS1 and LC-2/ad – CCDC6-RET). The library was sequenced using the Ion PGM™ system and analyzed with the AmpliSeq™ RNA Lung Fusion workflow in Ion Reporter™. Analysis showed that the positive control sample contained all expected fusions and control genes and reported zero false positives fusions. This multiplexed fusion transcript targeted
sequencing solution is currently being validated by all members of the
OncoNetwork Consortium who will test lung cancer tissue samples that have been well characterized by FISH, real-time PCR, IHC, and/or massarray. Initial results from OncoNetwork Consortia members reveal 100% concordance between the AmpliSeq™ RNA Lung Fusion panel and FISH in 25 lung tissue samples.
