engleski

Molecular diagnostics of severe congenital neutropenia and Shwachman-Diamond syndrome in seven Croatian families

Introduction: Severe congenital neutropenia (SCN) is an autosomal dominant disease characterized by chronic neutropenia which leads to severe recurrent bacterial or mycotic infections. Shwachman- Diamond syndrome (SDS) is an autosomal recessive disease that is characterized by exocrine pancreatic insufficiency, dysfunction of the bone marrow and predisposition to develop myelodysplastic syndrome or leukemia. Objective: The objective was to confirm clinical diagnosis in several Croatian families at genetic level by analyzing their DNA samples. Methods: 20 samples of genomic DNA were analyzed: 7 samples of patients with clinical diagnosis of SCN or SDS and 13 samples of their family members. Coding regions of ELANE and SBDS gene were amplified by PCR with specific primers. PCR fragments were bidirecty sequenced using Big Dye® Terminator v3.1 Cycle Sequencing Kit and Applied Biosystems 3130xl Genetic analyzer. Results: In all 4 patients with SCN, mutations were found in ELANE gene. These mutations were de novo, because in their parents' samples mutations were not found. In all 3 patients with SDS, mutations were found on both alleles, which were inherited from their parents. Two new variants were found, one in ELANE gene (c.213C>G ; p.Cys71Trp) and one in SBDS gene (c.41A>G ; p.Asn14Ser). Both variants were analyzed with PolyPhen-2 software and were predicted as probably damaging. Conclusion: Mutations in ELANE gene were reported in 38-80% of individuals with SCN, and in SBDS gene in more that 90% of individuals with SDS. In our case, clinical diagnoses of SCN or SDS were confirmed in all analyzed patients.

severe congenital neutropenia; Shwachman-Diamond syndrome; SBDS gene; ELANE gene; DNA sequencing

nije evidentirano