Desenvolvimento de um painel multiplex de qf-pcr para triagem das principais aneuploidias presentes na população

Abstract

Chromosomal abnormalities, especially aneuploidies, are the leading cause of intellectual disability in the human population, accounting for approximately 15% of congenital malformations. Viable human aneuploidies occur in chromosomes 13, 18, 21, X, and Y, and are associated with physical and cognitive limitations. The aim of this study was to develop, in silico, a multiplex QF-PCR panel to detect aneuploidies in pre- and postnatal samples. A literature review was conducted using PubMed with specific search terms related to aneuploidy assays by capillary electrophoresis, including chromosomes 13, 18, 21, X, and Y, as well as primer sequences and detailed methodologies. From the initial 281 articles, 22 were selected. The in silico analysis of primer sequences was performed using IGV, NCBI, and IDT, confirming the panel’s feasibility. Of the 90 initial sequences, 21 primers were approved and distributed across the target chromosomes. Primer interaction analyses indicated a need to divide them into two pools to reduce nonspecific annealing. Markers were arranged by size range and fluorescence channels, facilitating their application in capillary electrophoresis. The in-silico multiplex QF-PCR panel developed here requires in vitro validation to confirm its effectiveness under real-world conditions. This approach may foster advances in the molecular diagnosis of aneuploidies, providing a faster and more reliable method with potential for clinical application and future research.