La tecnologia del microchip: Una strategia avanzata per la caratterizzazione molecolare delle mutazioni più frequentemente associate a craniostenosi

Craniosynostosis, the premature fusion of one or more sutures of the skull, is a very heterogeneous group of disorders, in the etiology of which genetics play an important role. Considered as one of the most common congenital defect, it occurs with a prevalence of 1 out 2500 live births. Untreated progressive craniosynostosis leads to inhibition of brain growth and increased intracranial and intraorbital pressure. Due to the high heterogeneity of phenotypic expressions, molecular characterization of genes of interest may be very helpful in establishing a more accurate diagnostic evaluation. For the identification of 10 common mutations (S252W, P253R, F276V, C278F, C342R, C342Y, C342S, A344P, S347C e P250R) in fibroblast growth factor receptor 2 (FGFR2) and fibroblast growth factor receptor 3 (FGFR3) genes, we developed a microelectronic microchip assay. We set up conditions for polymerase chain reaction (PCR) multiplexing format coupled with serial addressing of the PCR products and serial probe hybridization for mutations in the FGFR genes. Analytical protocols for the detection of each mutation were optimized. The final version of the craniosynostosis microchip allowed the identification of all the mutations in FGFR2 and FGFR3 genes on a single pad per patient. The analysis was performed in a cohort of 159 patients with various craniosynostosis syndromes. The microchip strategy allows a rapid and specific identification of the common mutations of FGFR genes involved in craniosynostosis. The precocious molecular characterization of these genes in newborns or infants represents an effective tool for the medical and surgical management of these anomalies.