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 largely undocumented. Molecular means can be provided for the diagnosis of CF in this population. The development of a simple and rapid molecular diagnostic approach which can be used for diagnosis of CF, determination of the carrier status and investigation of the genotype/phenotype relationships in Saudi individuals. Specifically, the assays presented here should enable the detection of mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene found to comprise some 70% of CF alleles amongst Saudi patients. These mutations are: 3120+1G → A, N1303K, 1548delG, 2043delG, I1234V and 406-2A → G. Patients investigated & methods: Fifteen children with CF and their parents from 12 Saudi families from the Eastern Region of Saudi Arabia. Amplification by polymerase chain reaction (PCR) of the respective regions of the CFTR gene, digestion with appropriate restriction enzyme and agarose gel electrophoresis. Results: All mutations known from sequencing were reliably detected in the patients and their parents, and the carrier status was accurately determined in each case. Conclusion: This approach is simple and highly accurate, applicable for routine diagnosis, suitable for detection of the carrier status and obviates the need for mutation scanning by sequencing in the majority of cases.