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Updated: Mar 24, 2026
Targeted Next-generation Sequencing and Bioinformatics Pipeline to Evaluate Genetic Determinants of Constitutional Disease
Published on: April 4, 2018
Efficient Targeted Next Generation Sequencing-Based Workflow for Differential Diagnosis of Alport-Related Disorders
Gábor Kovács1, Tibor Kalmár1, Emőke Endreffy1
1University of Szeged, Faculty of Medicine, Department of Pediatrics and Pediatric Health Center, Szeged, Hungary.
Insights
Early genetic testing for Alport syndrome (AS) mutations in COL4A3-A5 genes is crucial for timely treatment. This study presents an efficient workflow for diagnosing AS and related disorders, potentially eliminating the need for renal biopsies.
Area of Science:
- Genetics
- Nephrology
- Molecular Biology
Background:
- Alport syndrome (AS) is a genetic kidney disease caused by mutations in type IV collagen genes.
- Early diagnosis of AS is vital for effective treatment and improved patient outcomes.
- Current diagnostic methods can be time-consuming, delaying critical therapeutic interventions.
Purpose of the Study:
- To develop and validate an efficient next-generation sequencing workflow for simultaneous analysis of COL4A3, COL4A4, and COL4A5 genes.
- To establish a unified diagnostic terminology and workflow for Alport syndrome and related conditions.
- To demonstrate the utility of molecular genetic analysis in diagnosing AS and potentially obviating the need for renal biopsy.
Main Methods:
- Next-generation sequencing (NGS) workflow for simultaneous analysis of COL4A3-A5 genes.
- Application of the workflow to three individuals and fourteen families with suspected Alport syndrome.
- Identification and characterization of mutations in affected individuals.
Main Results:
- Successfully identified causative mutations in all investigated Alport syndrome cases.
- Discovered 14 novel mutations within the Hungarian cohort.
- Demonstrated the workflow's effectiveness across X-linked, autosomal, and Alport-related diseases.
- Confirmed the potential to replace renal biopsy with genetic testing in diagnosed families.
Conclusions:
- The developed NGS workflow provides an efficient and comprehensive method for diagnosing Alport syndrome and related disorders.
- Molecular genetic analysis is a powerful tool for early and accurate diagnosis, guiding therapeutic decisions.
- This approach simplifies diagnosis and may render renal biopsies unnecessary, improving patient management and reducing healthcare costs.
Abstract:
Alport syndrome (AS) is an inherited type IV collagen nephropathies characterized by microscopic hematuria during early childhood, the development of proteinuria and progression to end-stage renal disease. Since choosing the right therapy, even before the onset of proteinuria, can delay the onset of end-stage renal failure and improve life expectancy, the earliest possible differential diagnosis is desired. Practically, this means the identification of mutation(s) in COL4A3-A4-A5 genes. We used an efficient, next generation sequencing based workflow for simultaneous analysis of all three COL4A genes in three individuals and fourteen families involved by AS or showing different level of Alport-related symptoms. We successfully identified mutations in all investigated cases, including 14 unpublished mutations in our Hungarian cohort. We present an easy to use unified clinical/diagnostic terminology and workflow not only for X-linked but for autosomal AS, but also for Alport-related diseases. In families where a diagnosis has been established by molecular genetic analysis, the renal biopsy may be rendered unnecessary.
