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BRIEF RESEARCH REPORT article
Front. Genet.
Sec. Genetics of Common and Rare Diseases
Volume 15 - 2024 |
doi: 10.3389/fgene.2024.1493295
Genewise detection of variants in MEFV gene using nanopore sequencing
Provisionally accepted- 1 Institute of Molecular Biology, Armenian National Academy of Sciences, Yerevan, Armenia
- 2 Russian-Armenian (Slavonic) University, Yerevan, Yerevan, Armenia
- 3 Armenian Bioinformatics Institute (ABI), Yerevan, Armenia
- 4 Department of Rheumatology, Yerevan State Medical University after Mkhitar Heratsi (YSMU), Yerevan, Armenia
- 5 Department of Rheumatology, “Mikaelyan” Institute of Surgery, Yerevan, Armenia
- 6 Nairi Medical Center, Yerevan, Armenia
Familial Mediterranean Fever (FMF) is a genetic disorder with complex inheritance patterns and genotype-phenotype associations, and it is highly prevalent in Armenia. FMF typically follows an autosomal recessive inheritance pattern (OMIM: 249100), though it can occasionally display a rare dominant inheritance pattern with variable penetrance (OMIM։134610). The disease is caused by mutations in the MEFV gene, which encodes the pyrin protein. While the 26 most prevalent mutations account for nearly 99% of all FMF cases, more than 60 pathogenic mutations have been identified. In this study, we aimed to develop an affordable nanopore sequencing method for fulllength MEFV gene mutation detection to aid in the diagnosis and screening of FMF. PAGE \* Arabic \* MERGEFORMAT We employed a multiplex amplicon sequencing approach, allowing for the processing of up to 12 samples on both Flow cells and Flongle flow cells. The results demonstrated near-complete concordance between nanopore variant calling and qPCR genotypes. Moreover, nanopore sequencing identified additional variants, which were confirmed by whole exome sequencing. Additionally, intronic and UTR variants were detected.Our findings demonstrate the feasibility of full-gene nanopore sequencing for detecting FMFassociated pathogenic variants. The method is cost-effective, with costs comparable to those of the qPCR test, making it particularly suitable for settings with limited laboratory infrastructure. Further clinical validation using larger sample cohorts will be necessary.
Keywords: Familial Mediterranean Fever, FMF, MEFV, nanopore sequencing, Variant detection, Genetic Testing
Received: 08 Sep 2024; Accepted: 11 Nov 2024.
Copyright: © 2024 Ghukasyan, Khachatryan, Sirunyan, Minasyan, Hakobyan, Chavushyan, Hayrapetyan, Ghazaryan, Martirosyan, Gohar, Vardanyan, Mukuchyan, Zakharyan and Arakelyan. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Roksana Zakharyan, Russian-Armenian (Slavonic) University, Yerevan, 0051, Yerevan, Armenia
Arsen Arakelyan, Institute of Molecular Biology, Armenian National Academy of Sciences, Yerevan, 0014, Armenia
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