Comprehensive detection of alterations in acute lymphoblastic leukemia: a rapid and accurate novel approach.

Acute lymphoblastic leukemia (ALL) is a prevalent childhood cancer with high cure rate, but poses a significant medical challenge in adults and relapsed patients. Philadelphia-like acute lymphoblastic leukemia (Ph-like ALL) is a high-risk subtype, with approximately half of cases characterized by overexpression and frequent concomitant deletions. To address the need for efficient, rapid, and cost-effective detection of alterations, we developed a novel RT-qPCR technique combining SYBR Green and highresolution melting analysis on a single plate. The method successfully identified expression, fusions, and and variants, achieving a 100% sensitivity and specificity. Application of this method across 61 samples revealed that 24.59% exhibited overexpression, predominantly driven by (73.33%). High Resolution Melting analysis unveiled concurrent or variants in 8.19% of samples, as well as a dynamic nature of alterations during disease progression. Overall, this approach provides an accurate identification of alterations, enabling improved diagnostic and facilitating therapeutic decision-making.