A dust collection box and a robot vacuum cleaner are disclosed. The dust collection box is used in the robot vacuum cleaner. The dust collection box includes a box body, a filter holder arranged in the box body, a dust separation structure configured to block dust, and a filter. The box body defines an air inlet and an air outlet. The filter holder defines an opening in a side surface thereof. The dust separation structure is arranged in the box body and located at an air inlet side of the opening. The filter is arranged to the filter holder and located at an air outlet side of the opening.