Sex identification in birds through molecular methods is an important tool for the management and preservation of species. Advances in real-time PCR-based techniques overcome some limitations of the more classical molecular analysis methodologies. Here, we describe a new approach, based on high-resolution melting (HRM) curve analysis of the CHD1 gene, for avian gender identification. This method was successfully applied to carry out sexual differentiation based on melting curve patterns in common quail (Coturnix c. coturnix) and Japanese quail (Coturnix c. japonica). We clearly demonstrate the efficacy of a simple HRM assay for a rapid and efficient gender differentiation of these subspecies and propose this methodology as a valuable addition to expand the applicability of real-time PCR-based technology in avian molecular sexing.