CLINICAL UTILITY OF A HYBRID QPCR-SEQUENCING STRATEGY IN MRD MONITORING: APPROACH INCLUDING BIOINFORMATICS

Abstract

AML develops by a hierarchical architecture of founder and subclonal mutations, which contributes to significant inter- and intra-patient heterogeneity, according to extensive sequencing studies. The need for improved biomarkers that can direct risk-adapted treatment plans outside of traditional morphologic evaluation is highlighted by this complexity. Data processing, variant calling, error correction, and longitudinal tracking of clonal dynamics are all essential parts of this hybrid strategy, which also incorporates complex bioinformatics processes. To improve the specificity and sensitivity of MRD detection, methods including UMIs, error suppression algorithms, and classification models based on machine learning are utilised. To further improve the accuracy and clinical relevance of MRD status interpretation over time, bioinformatics technologies also allow for the integration of multi-modal data. Improving risk stratification, guiding personalised treatment decisions, and enabling early identification of recurrence are the clinical value of a hybrid qPCR-sequencing method. The study had some weaknesses, particularly in relation to variability with yield/quality of RNA; the susceptibility of RNA to contamination; and the necessity for precise optimisation of PCR parameters. The restrictions resulting from the limitations of this laboratory membrane were resolved through rigorous discipline to laboratory practices and through methodological technical controls that were established in accordance with strict laboratory safety requirements, which increased the credibility of the scientific data. In summary, this study confirms RT-qPCR as an extremely sensitive, specific, and reproducible methodology for the identification of the BCR-ABL fusion gene. Due to its ability to give consistent and dependable results, RT-qPCR is an essential resource for usage in diagnosing and monitoring hematologic aberrations related to the BCR-ABL fusion gene. Advantages of using RT-qPCR in high quality materials, along with standard assay protocols will result in optimal assay performance.