Abstract For the last decades vibration based damage detection of engineering structures has become an important issue for maintenance operations on transport infrastructure. Research in vibration based structural damage detection has been rapidly expanding from classic modal parameter estimation to modern operational monitoring. Since structures are subject to unknown ambient excitation in operation conditions, all estimates from the finite data measurements are of statistical nature. The intrinsic uncertainty due to finite data length, colored noise, non-stationary excitations, model order reduction or other operational influences needs to be considered for robust and automated structural health monitoring methods. In this paper, two subspace-based methods are considered that take these statistical uncertainties into account, first modal parameter and their confidence interval estimation for a direct comparison of the structural states, and second a statistical null space based damage detection test that completely avoids the identification step. The performance of both methods is evaluated on a large scale progressive damage test of a prestressed concrete road bridge, the S101 Bridge in Austria. In an on-site test, ambient vibration data of the S101 Bridge was recorded while different damage scenarios were introduced on the bridge as a benchmark for damage identification. It is shown that the proposed damage detection methodology is able to clearly indicate the presence of structural damage, if the damage leads to a change of the structural system.