This review outlines the evolution of molecular techniques used to understand the clinical and molecular heterogeneity of leukemia. These technical advances are reshaping disease classification, prognosis, and therapeutic strategies. The diagnostic journey has progressed from foundational methods like karyotyping and fluorescence in situ hybridization (FISH) to higher-resolution analyses. Techniques such as microarrays, optical genomic mapping (OGM), and next-generation sequencing (NGS) have enabled the identification of copy number variations, complex chromosomal rearrangements, and comprehensive mutational landscapes in key genes like FLT3, NPM1, and TP53. Furthermore, advanced methods have provided deeper insights. RNA-seq has revealed novel transcriptome components, including fusion genes and non-coding RNAs (ncRNA). The role of extrachromosomal DNA (ecDNA) in oncogene amplification and therapeutic resistance has recently been highlighted. Epigenetic dynamics are being explored through techniques like chromatin immunoprecipitation sequencing (ChIP-seq) and the assay for transposase-accessible chromatin (ATAC-seq). Integrating these complementary tools is crucial for identifying therapeutic vulnerabilities cost-effectively. Finally, targeted assays like RT-qPCR and Sanger sequencing remain essential for confirmatory diagnosis and the technical validation of whole-genome or transcriptome-wide assays.
Boggula et al. (Wed,) studied this question.