Wildlife-vehicle collisions (WVC) produce considerable costs in road traffic due to human fatalities as well as ecological and economic losses. Multiple mitigation measures have been developed over the past decades to separate traffic and wildlife, to warn humans, or to prevent wildlife from entering roads. Among these, wildlife warning reflectors (WWR) have been frequently implemented, although their effectiveness remains a subject of discussion due to conflicting study results. Here we present a literature review on the effectiveness of WWR for N = 76 studies, including their methodological differences, such as the type of WWR (model and color), study conditions, and study designs. We used boosted regression trees to analyse WVC-data addressed in the literature to compare WWR effectiveness depending on the study design, study conditions, effective study duration, length of the tested sections, time period of the study, data source, reflector type, and animal species. Our analyses revealed no clear evidence for the effectiveness of WWR in preventing WVC. Instead, our meta-analysis showed that most studies indicating significant effects of WWR on the occurrence of WVC may be biased due to insufficiencies in study design and/or the approach of WVC data acquisition. Our computation of log response ratios (LRRWVC) showed that only studies applying a before-after (BA) design concluded that WWR were effective. Moreover, BRT modeling revealed that only studies of <12 months effective study duration and <5 km test site length indicated that WWR might lower WVC. Based on the vulnerability to confounding factors of WWR-study designs applied in the past, this review suggests the standardization of study conditions, including a before-after control-impact (BACI) or a cross-over study design with spatial and temporal control sections, a minimum test site length and a minimum study duration.