Development and evaluation of a reverse dot blot assay for the simultaneous detection of six common Chinese G6PD mutations and one polymorphism

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common inherited disorder worldwide including southern China. We developed and validated a reverse dot blot (RDB) assay for the rapid and simultaneous genotyping of six mutations (c.95A>G, c.871G>A, c.1004C>T, c.1024C>T, c.1376G>...

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Bibliographic Details
Published in:Blood cells, molecules, & diseases molecules, & diseases, 2008-07, Vol.41 (1), p.17-21
Main Authors: Li, Liang, Zhou, Yu-Qiu, Xiao, Qi-Zhi, Yan, Ti-Zhen, Xu, Xiang-Min
Format: Article
Language:English
Subjects:
Asian Continental Ancestry Group - genetics
Blotting, Southern - methods
Exons
Female
Genotype
Glucose-6-phosphate dehydrogenase deficiency (G6PD)
Glucosephosphate Dehydrogenase - genetics
Glucosephosphate Dehydrogenase Deficiency - diagnosis
Humans
Male
Multiplex polymerase chain reaction (M-PCR)
Oligonucleotide Probes
Point Mutation
Point mutations
Polymerase Chain Reaction - methods
Polymorphism, Genetic
Reverse dot blot (RDB)
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Summary:Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common inherited disorder worldwide including southern China. We developed and validated a reverse dot blot (RDB) assay for the rapid and simultaneous genotyping of six mutations (c.95A>G, c.871G>A, c.1004C>T, c.1024C>T, c.1376G>T and c.1388G>A), that were common mutations in the Chinese G6PD deficiency population, and one polymorphism (c.1311C>T). Reliable genotyping of wild-type and mutant genomic DNA samples was achieved by means of a test strip onto which allele-specific oligonucleotide probe lines are fixed in parallel. This method involves a multiplex PCR amplification of three fragments in the G6PD target sequence and a manual hybridization/detection protocol. The entire procedure starting from blood sampling to the identification of mutations requires less than 6 h. The diagnostic reliability of this reverse dot blot assay was evaluated on 207 pre-typed samples by using direct DNA sequence analysis in a blind study. The reverse dot blot typing was in complete concordance with the reference method. The reverse dot blot assay was proved to be a simple, rapid, highly accurate, and cost-effective method to identify common G6PD mutations in Chinese population.
ISSN:1079-9796
1096-0961
DOI:10.1016/j.bcmd.2008.01.007