Autor(es): Martins, Sara ; Santos, Maria João ; Teixeira, Márcia ; Diogo, Luisa ; Macário, Maria do Carmo ; Pedro Marques, João ; Fonseca, Pedro ; Grazina, Manuela
Data: 2023
Identificador Persistente: https://hdl.handle.net/10316/105249
Origem: Estudo Geral - Universidade de Coimbra
Projeto/bolsa: info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UID/PT;
Assunto(s): High-resolution melting; LHON; idebenone; mtDNA variant; real-time PCR; theranostics
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. Ninety-four 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.
This work was financed by the European Regional Development Fund (ERDF), through the Centro 2020 Regional Operational Programme under project CENTRO-01-0145-FEDER-000012-N2323 (HealthyAging2020) and through the COMPETE 2020 – Operational Programme for Competitiveness and Internationalisation and Portuguese national funds via FCT – Fundação para a Ciência e a Tecnologia, under projects POCI-01-0145-FEDER-007440 (Strategic Project), UID/NEU/04539/2019, Pest-C/SAU/LA0001/2013e2014 and doctoral grant SFRH/BD/86622/2012. The LBioMiT was financed by Santhera Pharmaceuticals, allowing the implementation of the project “Providing free of charge complete genetic tests to Portuguese patients with a clinical and instrumental diagnosis of Optic Nerve Atrophy” (PI Professor Manuela Grazina).
