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

作者：Lu, Xiaomei^*; Hua, Liang; Zhang, Ting; Li, Siping; Fan, Xuejin; Peng, Qi; Li, Wenrui; Ye, Junqin; Long, Jianling; He, Xiaoguang

来源：Clinica Chimica Acta, 2013, 418: 45-49.

DOI：10.1016/j.cca.2012.12.023

• Glucose 6-phosphate dehydrogenase
• deficiency (G6PD)
• Reverse dot blot (RDB)
• Multiplex polymerase reaction (M-PCR)
• Point mutation
• Genetic screening

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.

• 出版日期2013-3-15
• 单位广州市妇女儿童医疗中心; 浙江大学