Long-read sequencing improves diagnostic rate in neuromuscular disorders

Area of Science:

• Genomics
• Molecular Biology
• Medical Genetics

Background:

• Short-read sequencing has limitations in diagnosing genetic diseases, including inaccurate genome assembly and inability to detect large structural or repetitive variants.
• Advancements in massive parallel sequencing have aided molecular diagnosis over the past two decades.

Purpose of the Study:

• To review the application of long-read sequencing (LRS) technologies, specifically PacBio SMRT and Oxford Nanopore Technology (ONT), in diagnosing genetic disorders.
• To highlight the advantages of LRS over short-read sequencing for detecting various types of genetic variants.

Main Methods:

• Review of recent studies utilizing PacBio SMRT and ONT sequencing methods.
• Analysis of variant detection capabilities of LRS in individuals with neuromuscular disorders.

Main Results:

• LRS methods successfully detect repeat expansions in novel disease-causing genes (e.g., ABCD3, NOTCH2NLC, RILPL1, PLIN4).
• LRS facilitates the identification of structural variants (e.g., in DMD) and single nucleotide variants in repetitive regions (e.g., TTN, NEB).
• LRS simplifies the characterization of D4Z4 repeats in DUX4, aiding Facioscapulohumeral muscular dystrophy (FSHD) diagnosis.

Conclusions:

• Long-read sequencing significantly increases the diagnostic rate for genetic diseases, particularly neuromuscular disorders.
• LRS technologies are crucial for overcoming the limitations of short-read sequencing and improving the detection of complex genetic variants.