PB1777: DEVELOPMENT OF NEXT GENERATION SEQUENCING PANELS FOR SCREENING FUSION GENES IN LEUKEMIA

Abstract

Background: Diagnosis of hematologic malignancies is based on multidisciplinary approaches including cytological, cytogenetic, and molecular genetic analyzes. Advances in cancer genomics have enabled the identification of recurrent mutations with the apparent prognostic value for various cancers. To date, hundreds of fusion genes resulting from chromosomal rearrangements have been identified in acute leukemia. Testing such mutations in leukemias is essential for personalized treatments. Many methods used in conventional clinical diagnosis could not fully detect fusion genes. Aims: To overcome the limitations of existing techniques, targeted next generation sequencing approach was tested in this study. Methods: The libraries of the regions containing the junctions of the fusion genes, formed by multiplex PCR-based panels, was deep-sequenced using Illumina platform, thereafter the results were analyzed by bioinformatics approaches. Results: Using AmpliSeq (Illumina) fusion panel adapted for 20 KMT2A fusion transcripts and DNA fragments mimicking these fusion transcripts, 19 amplicons could be obtained with mPZR. With NGS procedures and bioinformatics analyses, significant results were obtained for 17 amplicons, while few junction reads that precisely describe the fusion event were obtained for the remaining 3. In titration studies, it was shown that the fusion panel can detect the template DNA fragments in trace amounts up to 1 pg. Image:Summary/Conclusion: These results show that the fusion panel can be used in clinical diagnosis for the fusion transcripts in its scope, after testing on clinical specimens.