SOHO State of the Art Updates and Next Questions | Implementing Next-Generation Sequencing (NGS) in...

Next-generation sequencing (NGS) has transformed the diagnostic and prognostic landscape of acute myeloid leukemia (AML) by enabling the simultaneous detection of multiple clinically relevant mutations. However, access to this technology remains limited in many Latin American countries due to economic and infrastructural constraints. This study reports the implementation and validation of an NGS workflow for AML molecular profiling within the International Consortium on Acute Leukemias (ICAL), emphasizing feasibility and reproducibility across reference laboratories in Brazil. iSeq100 sequencers (Illumina) were donated to ICAL centers. Laboratory personnel received training through the American Society of Hematology's Visitor Training Program at Erasmus MC (Rotterdam). Genomic DNA from 15 AML patients enrolled in the ICAL-2015 study was analyzed using a custom amplicon-based panel targeting FLT3, NPM1, IDH1, IDH2, ASXL1, CEBPA, TP53, and RUNX1. Results were compared between the 2 Brazilian laboratories, a European reference laboratory, and conventional single-gene testing. NGS demonstrated positive interlaboratory concordance for variant allele frequencies (Pearson r = 0.72). Concordance between NGS and PCR-based methods reached 77%, with discordant cases involving FLT3-ITD detection and the assessment of ASXL1 mutations. Mutation frequencies were consistent with those reported in European and North American cohorts. The present study demonstrates that simplified NGS workflows can be successfully established and quality-controlled in resource-constrained settings through international collaboration, training, and standardized protocols.