Genewise detection of variants in MEFV gene using nanopore sequencing

Area of Science:

• Genetics
• Molecular Biology
• Bioinformatics

Background:

• Familial Mediterranean Fever (FMF) is a prevalent genetic disorder with complex inheritance.
• Mutations in the MEFV gene cause FMF, with over 60 identified pathogenic variants.
• Accurate and accessible diagnostic methods are crucial for FMF management.

Purpose of the Study:

• To develop a cost-effective nanopore sequencing method for full-length MEFV gene mutation detection.
• To improve FMF diagnosis and screening capabilities, particularly in resource-limited areas.

Main Methods:

• Multiplex amplicon sequencing using nanopore technology on Flow and Flongle cells.
• Comparison of nanopore variant calling with qPCR genotypes.
• Confirmation of identified variants using whole exome sequencing.

Main Results:

• Near-complete concordance between nanopore sequencing and qPCR for FMF genotypes.
• Nanopore sequencing identified additional pathogenic and non-coding (intronic, UTR) variants.
• The method proved feasible for comprehensive MEFV gene mutation detection.

Conclusions:

• Full-gene nanopore sequencing is a viable, cost-effective tool for detecting FMF-associated MEFV variants.
• This method offers a practical alternative to existing diagnostic tests, especially in settings with limited laboratory infrastructure.
• Further clinical validation with larger cohorts is recommended.