GenEye24: Novel rapid screening test for the top-3 Leber’s Hereditary Optic Neuropathy pathogenic sequence variants

• DOI: 10.1016/j.mito.2023.01.006
• Publication Date: 2023-01-28T05:38:53Z
• Authors: Sara Martins; Maria João Santos; Márcia Teixeira; Luísa Diogo; Maria do Carmo Ma...; João Pedro Marques; Pedro Fonseca; Manuela Grazina
• Keywords: 03 medical and health sciences; 0302 clinical medicine; Mutation; Humans; Optic Atrophy, Hereditary, Leber; Real-Time Polymerase Chain Reaction; DNA, Mitochondrial; 3. Good health

Leber's Hereditary Optic Neuropathy (LHON) has been mainly (90-95 %) associated to one of three variants: m.3460G>A, m.11778G>A, m.14484T>C. Herein, a screening method was developed for its detection, supporting clinical/therapeutics decision. It relies on real-time PCR with High-Resolution Melting (HRM) analysis. Variant classification is made using HRM Software and quality controls. A total of 101 samples were analyzed. All samples were correctly assigned: 58 wild-type, 35 positive for m.11778G>A, 6 positive for m.14484T>C, 2 positive for m.3460G>A. Results presented sensitivity = 1, specificity = 1, Positive Predictive Value = 1 and Negative Predictive Value = 1. A new Real-Time PCR/HRM screening method cost-efficient, simple, robust and quick, detecting LHON's top-3 is described.