PCR-BASED ANALYSIS OF CYSTIC-FIBROSIS MUTATIONS SPECIFIC FOR SAUDI PATIENTS

Objectives: The occurrence of cystic fibrosis (CF) in Saudi Arabia has been reported but the molecular basis of the disease still remains la rgely undocumented. Molecular means can be provided for the diagnosis of CF in this population. The development of a simple and rapid molecu lar diagnostic approach which can be used for diagnosis of CF, determi nation of the carrier status and investigation of the genotype/phenoty pe relationships in Saudi individuals. Specifically, the assays presen ted here should enable the detection of mutations in the cystic fibros is transmembrane conductance regulator (CFTR) gene found to comprise s ome 70% of CF alleles amongst Saudi patients. These mutations are: 312 0+1G -->A, N1303K, 1548delG, 2043delG, I1234V and 406-2A -->G. Patient s investigated & methods: Fifteen children with CF and their parents f rom 12 Saudi families from the Eastern Region of Saudi Arabia. Amplifi cation by polymerase chain reaction (PCR) of the respective regions of the CFTR gene, digestion with appropriate restriction enzyme and agar ose gel electrophoresis. Results: All mutations known from sequencing were reliably detected in the patients and their parents, and the carr ier status was accurately determined in each case. Conclusion: This ap proach is simple and highly accurate, applicable for routine diagnosis , suitable for detection of the carrier status and obviates the need f or mutation scanning by sequencing in the majority of cases.