Detection of Mediterranean Mutation in the Glucose-6-Phosphate Dehydrogenase Gene with Microarray Technique

Purpose: In this study, the type of mutations in Glucose-6-Phosphate Dehydrogenase deficient cases was defined using microarray technique. Methods: Twenty-five cases (17 male/8 female) with glucose-6-phosphate dehydrogenase deficiency determined by Beutler method were analyzed. DNA samples were isolated using Poncz principle to design hybridization probes for Gd-Mediterranean mutation. Subsequently, the 6(th) exon of glucose-6-phosphate dehydrogenase gene was amplified. After desalting, uncompounded primers were removed, the amplified polymerase chain reaction product was electronically addressed and denatured by NaOH. Following hybridization, the signals were automatically scanned, and evaluated by dedicated software. The NanoChip Molecular Biology Workstation(R) was standardized with 25 cases for Gd-Mediterranean mutation. Results: Twenty-five cases with glucose-6-phosphate dehydrogenase activity of 0-4.9 U/gHb were examined, and 13 homozygous, two heterozygous and 10 normal subjects for Gd-Mediterranean mutation were identified. These findings were consistent with the results obtained from RFLP method, which is considered the golden standard. Therefore, the Workstation(R) system has been found to be suitable to analyze the wild and mutant type of glucose-6-phosphate dehydrogenase in all cases. Conclusion: This research is the first study in which the microarray technique was utilized to detect Gd-Mediterranean that is the most seen glucose-6-phosphate dehydrogenase mutation in Turkey. The microarray technique allows large number of samples to be tested in a short period with no use of radioactive materials. In our study, genotypes were correctly identified for all subjects.