Glucose-6-phosphate dehydrogenase (G6PD) gene mutations detection by improved high-resolution DNA melting assay

Abstract

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common inherited disorder worldwide including southern China. G6PD gene mutations cause deficiency of the enzyme and a large spectrum of diseases. High-resolution DNA melting (HRM) assay was recently introduced as a rapid, inexpensive and effective method for genotyping. But there was a shortcoming of this method that hemizygous and homozygous genotypes were not easily distinguished from wild-types. Here we used improved HRM method for a small-scale screening of G6PD-deficient variants among people of Meizhou region. Then all amplicons were ascertained by direct DNA sequencing. These results indicated that HRM method was a major technical advance for G6PD mutations screening.

Rapid screening for sickle cell disease by polymerase chain reaction-high resolution melting analysis

“…Theoretically, adding one equivalent of reference DNA (wild-type) into homozygous samples produces heteroduplex DNA (heterozygotes), so that the curve profiles of the former and latter are able to be distinguished. This approach has been successfully verified in a previous study by our group (12).…”

Incidence and molecular characterization of Glucose‐6‐Phosphate Dehydrogenase deficiency among neonates for newborn screening in Chaozhou, China

“…In addition, two common double heterozygote mutants, G1376T/IVS XI t93c and G1388A/IVS XI t93c, are located in close proximity to one another. A previous study has reported that there may be some incorrect classification of these heterozygote mutants based on HRM analysis . Therefore, samples with these two mutations detected by HRM analysis were further confirmed by sequencing the segments that contained the specific sites (1376G>C, IVS XI t93c, and 1388G>A).…”

Clinical Significance of UGT1A1 Genetic Analysis in Chinese Neonates with Severe Hyperbilirubinemia

“…For the melting-curve shapes of homozygous mutations, which were similar to those of wild-type, homozygous mutations were detected by a modified HRM-analysis strategy as described our previous study. 16,21 All 58 case samples were correctly genotyped as compared with the sequence results. Therefore, we believed that HRM may be used as a rapid method for a large-scale investigation of UGT1A1 gene polymorphism.…”