Integrated Strategy for Fast and Automated Molecular Characterization of Genes Involved in Craniosynostosis

Abstract

Background: Craniosynostosis, the premature fusion of 1 or more sutures of the skull, is a common congenital defect, with a prevalence of 1 in 2500 live births. Untreated progressive craniosynostosis leads to inhibition of brain growth and increased intracranial and intraorbital pressure. The heterogeneity of clinical phenotypes and the overlap of the various associated syndromes render the correct diagnosis of the different craniosynostoses particularly difficult. Methods: To identify 10 common mutations in the genes for fibroblast growth factor receptors 2 and 3 (FGFR2 and FGFR3), we developed a microelectronic microchip assay that exploited the PCR multiplexing format and coupled it with serial addressing and probe hybridization on the same pad. For the molecular characterization of patients who tested negative in the microchip screening, we also developed conditions for denaturing HPLC (DHPLC) analysis of the most mutated regions of FGFR2 and FGFR3 and the entire coding region of the TWIST1 gene. Results: In our cohort of 159 patients with various craniosynostosis syndromes, mutations were found in

Cranial Ultrasound as a First-Line Imaging Examination for Craniosynostosis

“…Although craniosynostosis is the most frequent craniofacial anomaly, premature closure of cranial sutures is an uncommon condition. [1][2][3] Imaging studies rule out abnormal suture closure in most cases. In our study, as expected, the great majority of children (93.7%) had a negative radiograph.…”

“…[1][2][3] Craniosynostosis can present as an isolated finding or in association with various syndromes. Although an isolated single cranial suture closure usually causes only cosmetic deformity, 1 poor gross motor function and learning difficulties resulting from even a single suture synostosis have been reported.…”

Effectiveness of ultrasonographic evaluation of the cranial sutures in children with suspected craniosynostosis

“…Craniosynostosis, defined as a premature closure of one or more cranial sutures that prevents skull growth in the direction perpendicular to the suture plane, is the most frequent cranio-facial anomaly [1][2][3]. CS is a part of more than 180 syndromes and is associated with various skeletal dysmorphisms [9].…”

“…Craniosynostosis (CS) occurs in 4-6 of 10,000 live births [1][2][3]. Syndromatic craniosynostosis often involves multiple sutures.…”

Peptide-nucleic acid-mediated enriched polymerase chain reaction as a key point for non-invasive prenatal diagnosis of -thalassemia

“…3 Nevertheless, the enrichment of fetal sequences through gel elution is problematic and prone to contamination. As an alternative approach, we evaluated a microelectronic microchip technology that proved to be highly reliable in the identification of mutations in several disease genes [7][8][9][10][11] coupled with PNA-clamping of maternal wild-type allele to preferentially amplify fetal mutated sequences. PNAs are DNA analogs in which the ribosephosphate backbone is replaced by a peptide backbone.…”