Real time-PCR a diagnostic tool for reporting copy number variation and relative gene-expression changes in pediatric...

Area of Science:

• Molecular Diagnostics
• Genomics
• Transcriptomics

Background:

• Real-time polymerase chain reaction (RT-PCR) is a standard, cost-effective molecular diagnostic tool.
• Pediatric B-cell Acute Lymphoblastic Leukemia (ped B-ALL) diagnosis benefits from incorporating copy number variation (CNV) analysis alongside cytogenetics and clinical features for patient stratification.

Purpose of the Study:

• To develop a RT-PCR-based method for reporting CNVs and relative gene-expression changes.
• To establish a user-friendly and cost-effective diagnostic tool for genomic and transcriptomic alterations.
• To demonstrate the applicability of the method in pediatric B-cell Acute Lymphoblastic Leukemia (ped B-ALL) as a model.

Main Methods:

• Developed an iterative probability scoring method using RT-PCR.
• Applied the method to identify CNVs and gene-expression changes in ped B-ALL.
• Focused on genes of interest and CNV hotspots identified in a discovery phase.

Main Results:

• Successfully developed and validated a RT-PCR-based method for reporting CNVs and gene-expression changes.
• The method is cost-effective and user-friendly for identifying genomic and transcriptomic alterations.
• Demonstrated the potential for upfront patient stratification in ped B-ALL.

Conclusions:

• The proposed RT-PCR method offers a cost-effective solution for CNV and gene-expression analysis in diagnostics.
• This approach can be integrated into routine molecular diagnostics, particularly in resource-constrained environments.
• The method is adaptable for various diseases, enabling tailored diagnostic applications.