Grey mould is a fungal disease responsible for important crop losses in most winemaking regions worldwide. In the present study, we developed immunological techniques for the detection and quantification of Botrytis cinerea conidia germinative material to obtain additional information about epidemic’s development on the vineyard. The study was carried out in a northwest Spain vineyard during the grapevine flowering in 2017 and 2018. An aerobiological study was developed for the identification and quantification of Botrytis cinerea conidia. For the immunological analysis, we developed a specific protein extraction protocol and a standard curve to analyze the cyclone-trap samples by an indirect PTA-ELISA. The airborne B. cinerea protein concentrations showed a similar variation pattern than the airborne conidia concentrations in the atmosphere of the vineyard. We recorded a total of 5673 B. cinerea spores in 2017, and 7562 spores in 2018. Regarding the airborne protein concentrations, we detected 9.692 ng/m3 in 2017 and 7.715 ng/m3 in 2018. Based on the statistical influence of the considered variables, we developed a predictive model able to explain 40% of protein data variability. The resulting methodology based on the combination of immunological techniques and aerobiological monitoring leads to a more reasoned treatment schedule consistent with real phytosanitary vineyard conditions and a more effective responsiveness against the increasing variability associated to climate change on the crop-pathogen system.