Biomarkers.

Area of Science:

• Genetics
• Neuroscience
• Biochemistry

Background:

• Apolipoprotein E (ApoE) is crucial for lipid metabolism, glucose, and cholesterol homeostasis.
• APOE gene polymorphisms define six genotypes influencing metabolic function.
• APOE genotyping identifies susceptibility to late-onset Alzheimer's disease (AD), particularly with the APOE ε4 allele.

Purpose of the Study:

• To develop and validate a novel, simple, and affordable assay for Apolipoprotein E (APOE) genotyping.
• To assess the utility of this assay in identifying individuals at risk for neurodegenerative disorders, specifically late-onset Alzheimer's disease.
• To investigate APOE allelic frequencies in a Cuban population with neurodegenerative disorders.

Main Methods:

• Genomic DNA extraction from dried blood spots using Chelex-100.
• Development of allele-specific primers for APOE alleles (ε2, ε3, ε4) and internal controls.
• Multiplex real-time PCR combined with high-resolution melting analysis (HRMA) for APOE genotyping.

Main Results:

• The novel SUMASIGNAL APOE assay demonstrated 100% concordance with a commercial kit.
• Achieved low intra- and inter-assay coefficients of variation (<0.46%).
• The method can detect DNA concentrations as low as 5 ng/µL.

Conclusions:

• The SUMASIGNAL APOE assay is specific, simple, and cost-effective for APOE genotyping.
• This validated assay facilitates risk assessment and diagnosis for conditions like Alzheimer's disease.
• Enabled a pioneering study in Cuba on APOE allelic frequencies in neurodegenerative disease populations.