When used for the evaluation of drug efficacy against
Psoroptes ovis
, the diagnostic performance of different sampling strategies for a mite count reduction test (MCRT) remains unclear. In the present study, a novel simulation framework was constructed that accounted for relevant biological features of
P. ovis
infestations in cattle and that was parameterized with field data from 16 farms (154 animals). Second, this framework was applied to explore the impact of study specific factors (number of animals, number of sampled lesions, and number of scrapings per lesion) and biological factors (mite infestation intensity and size of lesions) on the diagnostic performance of MCRT. Its outcome provided a basis to determine the diagnostic performance of MCRT when it was applied according to the World Association for the Advancement in Veterinary Parasitology (WAAVP) and the European Medicine Agency (EMA) guidelines, and to formulate recommendations to ensure a good diagnostic performance of the MCRT. For both guidelines, the MCRT allowed to correctly detect (power 80%) reduced and normal efficacy when the therapeutic efficacy was <70%, and ≥95%, respectively. The results highlighted a reliable diagnostic performance of the MCRT when performed as recommended by WAAVP and EMA for the detection of normal drug efficacy. When used for the detection of reduced efficacy, therapeutic efficacies between 70% and 90% could not be detected with sufficient reliability. The diagnostic performance can be improved by increasing the total number of skin scrapings (increasing the number of animals, number of sampled lesions and/or number of samples per lesion). In order to help researchers and veterinarians to optimize the design of the MCRT to their field settings, the findings were translated into a simple tool.