In the present study, we describe the development and validation of a tNGS approach for the simultaneous detection of SNVs and copy number variations (CNVs) in genes associated with HPA (PAH, GCH1, PTS, QDPR, PCBD1 and DNAJC12) or useful for its differential diagnosis (SPR). Here, PCBD1 is linked to pulmonary arterial hypertension.