To meet consumer requirements for comfort in wooden structure construction, test mode methods (the environmental excitation method and impact excitation method) have been used to test six measuring points of the flooring in a two-story residential light-wood structure. The following tests were performed: the fundamental frequency test of floor structure under environmental excitation mode; the ball excitation dynamic vibration test of single and rhythmic running of a basketball and tennis ball under impact excitation mode; and the pedestrian dynamic vibration test of jump, single-step, steady walking, and rhythmic movement. The comfort analysis was validated based on the test results of peak value and effective value of fundamental frequency, acceleration, and speed. ANSYS was used to verify the calculation mode of the floor structure. Research showed that the fundamental frequencies of the building structure obtained through the calculation mode and the test mode were consistent, and both were higher than 4.5 Hz. The maximum measured acceleration peak value under the impact excitation mode was 407.2 mm/s2. The maximum speed peak value was 5.606 mm/s. The maximum acceleration effective value (RMS) was less than 450 mm/s2. The floor structure results met the building comfort requirements. The research has value in engineering applications, as it advances understanding concerning the vibration characteristics and comfort optimization of light-wood frame construction.