Precision Diagnosis of Wilson Disease Using a MultiGene Panel: Insights From a Prospective Cohort Study

Wilson disease (WD) is an autosomal recessive disorder of copper metabolism. Conventional genetic diagnostics are low-throughput and may miss intronic, structural, or phenocopy variants, leading to delayed or missed diagnoses. In this study, we evaluate the utility of a custom next-generation sequencing (NGS) panel targeting the full-length ATP7B gene and 10 additional copper metabolism-related genes in patients with clinically suspected WD.

We conducted a prospective cohort study of 144 individuals at our neurogenetic center. Variants identified by NGS were filtered and annotated with in silico tools and classified according to American College of Medical Genetics and Genomics guidelines. Confirmatory Sanger sequencing, multiplex ligation-dependent probe amplification, and reverse transcription PCR assays were performed as needed.

Genetic confirmation of WD was achieved in 129 of 144 patients (90%), including 80 typical (Leipzig score ≥4) and 49 atypical (score <4) cases. Ten novel ATP7B variants, including deep intronic, noncanonical splice, and copy number variants, were identified using this panel. Among 15 genetically unresolved cases, 6 harbored variants in other copper metabolism-related genes but no pathogenic ATP7B variants. Notably, 1 patient with a Leipzig score of 4 had been clinically diagnosed with WD for years but was reclassified as spinocerebellar ataxia type 12 after panel testing revealed only a heterozygous CP variant and a CAG repeat expansion in PPP2R2B.

Our comprehensive multigene NGS panel enables precise diagnosis of WD by detecting both classical and unconventional pathogenic variants, as well as distinguishing phenocopies. This improved diagnostic accuracy underscores the value of early genetic testing to guide timely intervention, especially in atypical or early-stage cases.