title: Diagnosis of Primary Ciliary Dyskinesia by a Targeted Next-Generation Sequencing Panel: Molecular and Clinical Findings in Italian Patients.
abstract: Primary ciliary dyskinesia (PCD) is a rare genetic disorder that alters mucociliary clearance, with consequent chronic disease of upper and lower airways. Diagnosis of PCD is challenging, and genetic testing is hampered by the high heterogeneity of the disease, because autosomal recessive causative mutations were found in 34 different genes. In this study, we clinically and molecularly characterized a cohort of 51 Italian patients with clinical signs of PCD. A custom next-generation sequencing panel that enables the affordable and simultaneous screening of 24 PCD genes was developed for genetic analysis. After variant filtering and prioritization, the molecular diagnosis of PCD was achieved in 43% of the patients. Overall, 5 homozygous and 27 compound heterozygous mutations, 21 of which were never reported before, were identified in 11 PCD genes. The DNAH5 and DNAH11 genes were the most common cause of PCD in Italy, but some population specificities were identified. In addition, the number of unsolved cases and the identification of only a single mutation in six patients suggest further genetic heterogeneity and invoke the need of novel strategies to detect unconventional pathogenic DNA variants. Finally, despite the availability of mutation databases and in silico prediction tools helping the interpretation of variants in next-generation sequencing screenings, a comprehensive segregation analysis is required to establish the in trans inheritance and support the pathogenic role of mutations.
authors.affiliation: Department of Biology, University of Padova, Padova, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Boaretto
authors.affiliation: Department of Women's and Children's Health, University of Padova, Padova, Italy.
authors.forename: Deborah
authors.initials: D
authors.lastname: Snijders
authors.affiliation: Department of Biology, University of Padova, Padova, Italy.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Salvoro
authors.affiliation: Department of Biology, University of Padova, Padova, Italy.
authors.forename: Ambra
authors.initials: A
authors.lastname: Spalletta
authors.affiliation: Department of Biology, University of Padova, Padova, Italy.
authors.forename: Maria Luisa
authors.initials: ML
authors.lastname: Mostacciuolo
authors.affiliation: Cystic Fibrosis and Respiratory Pediatric Center, Arnas Children Hospital, Palermo, Italy.
authors.forename: Mirella
authors.initials: M
authors.lastname: Collura
authors.affiliation: Department of Mother and Child Health, Salesi Children's Hospital, Ancona, Italy.
authors.forename: Salvatore
authors.initials: S
authors.lastname: Cazzato
authors.affiliation: Pediatric Pulmonology Unit, Istitute Giannina Gaslini, Genova, Italy.
authors.forename: Donatella
authors.initials: D
authors.lastname: Girosi
authors.affiliation: Pediatric Pulmonology Unit, Istitute Giannina Gaslini, Genova, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Silvestri
authors.affiliation: Pediatric Pulmonology Unit, Istitute Giannina Gaslini, Genova, Italy.
authors.forename: Giovanni Arturo
authors.initials: GA
authors.lastname: Rossi
authors.affiliation: Department of Women's and Children's Health, University of Padova, Padova, Italy.
authors.forename: Angelo
authors.initials: A
authors.lastname: Barbato
authors.affiliation: Department of Biology, University of Padova, Padova, Italy. Electronic address: giovanni.vazza@unipd.it.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Vazza
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D056444:Axonemal Dyneins / Q000235:genetics; D015415:Biomarkers; D002648:Child; D002675:Child, Preschool; D004252:DNA Mutational Analysis; D005260:Female; D056726:Genetic Association Studies; D005819:Genetic Markers; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D007223:Infant; D007558:Italy; D007619:Kartagener Syndrome / Q000175:diagnosis* / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D010641:Phenotype; D055815:Young Adult
chemical_list: D015415:Biomarkers; D005819:Genetic Markers; D056444:Axonemal Dyneins