Determination of Cell-Free DNA (cfDNA) Methylation by Methylation-Sensitive High-Resolution Melting (MS-HRM) in Human Plasma for the Diagnosis of Gestational Diabetes Mellitus (GDM)

Abstract

Considerable interest has focused on cell-free DNA (cfDNA) methylation for noninvasive prenatal diagnosis. We investigated maternal plasma DNA methylation using a simple protocol. Eighty plasma samples from pregnant women were separated for cfDNA methylation determination, including 13 from patients with gestational diabetes mellitus (GDM). Four CpG sites of the LHX3 gene were detected. The results show that methylation-sensitive high-resolution melting (MS-HRM) may be used for semi-quantitative determination of cfDNA methylation. There are extensive cfDNA methylation modifications at different gestational ages and different patterns were observed at different CpG sites. Hypermethylation was observed in the second trimester of pregnancy and GDM. Significant differences were observed in the two CpG sites in LHX3 between those with GDM and in the second trimester. It is suggested that cfDNA methylation and its level may be biomarkers for noninvasive prenatal diagnosis.