Long-read sequencing for molecular diagnostics in constitutional genetic disorders

Area of Science:

• Genomics
• Molecular Diagnostics
• Bioinformatics

Background:

• Long-read sequencing (LRS) technology has existed for over a decade but faced slow adoption due to high costs and error rates.
• Recent advancements have led to highly accurate LRS with reduced costs, increasing its potential.
• Short-read sequencing has limitations in detecting structural variations, repetitive regions, and phasing haplotypes.

Purpose of the Study:

• To evaluate the potential of LRS in molecular diagnostics for constitutional disorders.
• To contrast the advantages of LRS platforms (PacBio, Oxford Nanopore) with current standards of care.
• To critically assess the clinical utility and analytical validity of LRS in diagnostic settings.

Main Methods:

• Review of current literature on LRS applications and limitations.
• Comparison of LRS platforms (PacBio, Oxford Nanopore) for molecular diagnostics.
• Analysis of LRS capabilities in detecting structural variations, repetitive regions, haplotype phasing, and base modifications.

Main Results:

• LRS demonstrates significant advantages over short-read sequencing in resolving complex genomic regions and structural variations.
• Successful applications show LRS can identify molecular diagnoses missed by short-read sequencing.
• PacBio and Oxford Nanopore are key LRS platforms with distinct applications in diagnostics.

Conclusions:

• LRS has the potential to enhance molecular diagnostics for constitutional disorders by overcoming short-read limitations.
• Validation through larger cohort studies and development of clinical standards are necessary to establish LRS's diagnostic yield and utility.
• Continued advancements and critical evaluation are crucial for integrating LRS into routine diagnostic settings.