1. A reverse dot blot assay for the expanded screening of eleven Chinese G6PD mutations.
- Author
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Lu X, Hua L, Zhang T, Li S, Fan X, Peng Q, Li W, Ye J, Long J, and He X
- Subjects
- Adolescent, Adult, Child, Child, Preschool, China, Female, Humans, Immunoblotting, Infant, Infant, Newborn, Male, Polymerase Chain Reaction, Young Adult, Asian People genetics, DNA Mutational Analysis methods, Glucosephosphate Dehydrogenase Deficiency genetics
- Abstract
Background: Glucose-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., Methods: We 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., Results: The test had a detection rate of 95.8%, validated by direct sequencing in a blind study with 100% concordance., Conclusions: The 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., (Copyright © 2013 Elsevier B.V. All rights reserved.)
- Published
- 2013
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