A reverse dot blot assay for the expanded screening of eleven Chinese G6PD mutations

BackgroundGlucose-6-phosphate dehydrogenase (G6PD) deficiency is a multiethnic inherited disease with a particularly high prevalence in tropical and subtropical regions including southern China. A convenient and reliable method is required to detect common G6PD mutations in the Chinese population.MethodsWe developed a reverse dot blot (RDB) assay for the expanded screening of eleven mutations (c.95A>G, c.392G>T, c.871G>A, c.1004C>T, c.1004C>A, c.1024C>T, c.1360C>T, c.1376G>T, c.1381G>A, c.1387C>T, c.1388G>A). The method consists of a single-tube multiplex PCR amplification of four fragments in the G6PD target sequence of wild-type and mutant genomic DNA samples followed by hybridization to a test strip containing allele-specific oligonucleotide probes. We applied our method to a group of 213 unrelated Chinese patients.ResultsThe test had a detection rate of 95.8%, validated by direct sequencing in a blind study with 100% concordance.ConclusionsThe results demonstrate that our reverse dot blot assay is an easy, reliable, high-yield and cost-effective method for genetic screening to identify G6PD patients and carriers among the Chinese population.

► Glucose-6-phosphate dehydrogenase deficiency mainly affects southern China.
► A reverse dot blot assay is developed for the expanded screening of eleven mutations.
► The test had a detection rate of 95.8% in 213-unrelated-patient group.
► It was validated by direct sequencing in a blind study with a complete concordance.