Molecular characterization of G6PD mutations identifies new mutations and a high frequency of intronic variants in Thai females
2
Issued Date
2023-11-20
Resource Type
eISSN
19326203
Scopus ID
2-s2.0-85177069550
Pubmed ID
37967096
Journal Title
PLoS ONE
Volume
18
Issue
11 November
Rights Holder(s)
SCOPUS
Bibliographic Citation
PLoS ONE Vol.18 No.11 November (2023)
Suggested Citation
Chamchoy K., Sudsumrit S., Wongwigkan J., Petmitr S., Songdej D., Adams E.R., Edwards T., Leartsakulpanich U., Boonyuen U. Molecular characterization of G6PD mutations identifies new mutations and a high frequency of intronic variants in Thai females. PLoS ONE Vol.18 No.11 November (2023). doi:10.1371/journal.pone.0294200 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/91210
Title
Molecular characterization of G6PD mutations identifies new mutations and a high frequency of intronic variants in Thai females
Other Contributor(s)
Abstract
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an X-linked enzymopathy caused by mutations in the G6PD gene. A medical concern associated with G6PD deficiency is acute hemolytic anemia induced by certain foods, drugs, and infections. Although phenotypic tests can correctly identify hemizygous males, as well as homozygous and compound heterozygous females, heterozygous females with a wide range of G6PD activity may be misclassified as normal. This study aimed to develop multiplex high-resolution melting (HRM) analyses to enable the accurate detection of G6PD mutations, especially among females with heterozygous deficiency. Multiplex HRM assays were developed to detect six G6PD variants, i.e., G6PD Gaohe (c.95A>G), G6PD Chinese-4 (c.392G>T), G6PD Mahidol (c.487G>A), G6PD Viangchan (c.871G>A), G6PD Chinese-5 (c.1024C>T), and G6PD Union (c.1360C>T) in two reactions. The assays were validated and then applied to genotype G6PD mutations in 248 Thai females. The sensitivity of the HRM assays developed was 100% [95% confidence interval (CI): 94.40%–100%] with a specificity of 100% (95% CI: 88.78%–100%) for detecting these six mutations. The prevalence of G6PD deficiency was estimated as 3.63% (9/248) for G6PD deficiency and 31.05% (77/248) for intermediate deficiency by phenotypic assay. The developed HRM assays identified three participants with normal enzyme activity as heterozygous for G6PD Viangchan. Interestingly, a deletion in intron 5 nucleotide position 637/638 (c.486-34delT) was also detected by the developed HRM assays. G6PD genotyping revealed a total of 12 G6PD genotypes, with a high prevalence of intronic variants. Our results suggested that HRM analysis-based genotyping is a simple and reliable approach for detecting G6PD mutations, and could be used to prevent the misdiagnosis of heterozygous females by phenotypic assay. This study also sheds light on the possibility of overlooking intronic variants, which could affect G6PD expression and contribute to enzyme deficiency.