Speech reception thresholds were measured to investigate the influence of a room on speech segregation between a spatially separated target and interferer. The listening tests were realized under headphones. A room simulation allowed selected positioning of the interferer and target, as well as varying the absorption coefficient of the room internal surfaces. The measurements involved target sentences and speech-shaped noise or 2-voice interferers. Four experiments revealed that speech segregation in rooms was not only dependent on the azimuth separation of sound sources, but also on their direct-to-reverberant energy ratio at the listening position. This parameter was varied for interferer and target independently. Speech intelligibility decreased as the direct-to-reverberant ratio of sources was degraded by sound reflections in the room. The influence of the direct-to-reverberant ratio of the interferer was in agreement with binaural unmasking theories, through its effect on interaural coherence. The effect on the target occurred at higher levels of reverberation and was explained by the intrinsic degradation of speech intelligibility in reverberation.