The windowed interpolation DFT methods have been utilized to estimate the parameters of a single frequency and multi-frequency signal. Nevertheless, they do not work well for the real-valued sinusoids with closely spaced positive- and negative- frequency. In this paper, we describe a novel three-point windowed interpolation DFT method for frequency measurement of real-valued sinusoid signal. The exact representation of the windowed DFT with maximum sidelobe decay window (MSDW) is constructed. The spectral superposition of positive- and negative-frequency is considered and calculated to improve the estimation performance. The simulation results match with the theoretical values well. In addition, computer simulations demonstrate that the proposed algorithm provides high estimation accuracy and good noise suppression capability.