Botrytis cinerea, the causative agent of Botrytis bunch rot, is one of the most economically significant diseases of grapevines worldwide. The use of fungicides to control B. cinerea is an integral part of disease management in grapevines. Fungicide resistance in B. cinerea for all current single-site fungicides is geographically widespread (Leroux et al 1999, Latorre et al 2012). In this report, isolates (N = 504) were collected between 2020 and 2024 from 53 vineyards across Western Australia, South Australia, Victoria, New South Wales and the Australian Capital Territory. The isolates were screened on discriminatory concentrations of three chemical groups, anilinopyrimidines (group 9), phenylpyroles (group 12) and ketoreductase inhibitors (group 17). Resistance frequencies were 14.5, 9.7 and 8.7% for group 9, 12 and 17, respectively. All isolates resistant to group 9 and 17 were genotyped for the target genes Pos5 or Mdl1 (group 9) and Erg27 (group 17). In Pos5 and Mdl1, the changes V273I, P293S, P319A, G408V or L412F/V and E407K were identified, respectively. In Erg27, the changes F412C/I/S/V were found. Quantitative PCR based assays were designed and optimised to detect L412F/V (Pos5) and F412S (Erg27) mutations for use in lab-based high throughput and field-based rapid qPCR testing systems (Sosnowski et al. 2023). Rotorod spore traps were deployed nationally during the 2023/2024 and 2024/2025 seasons and rods were screened for the L412F mutation in the lab-based high throughput qPCR system. Testing results showed detection of L412F at sites in WA, SA, NSW, Vic and Tas. A field-based method was developed, adapted from a previous report that detected G143A (group 11, CytB) in Blumeria graminis f. sp. triciti on barley leaves in situ (Dodhia et al. 2021). Within a vineyard setting, infected berries were sampled from different blocks and hand ground in a buffer for the quick extraction of DNA. The resulting mixture was used as a template for qPCR in a mobile and lightweight qPCR instrument. The frequencies of L412F and F412S were 8-90% and 4-95%, respectively. Overall, the screening results have provided valuable information on the resistance status of B. cinerea in Australian vineyards and could assist in improving resistance management strategies. The combination of spore trapping and high throughput lab-based qPCR testing provides a means to monitor resistance throughout the growing season while the in-field qPCR method provides a quick and simple early detection approach in vineyards with known control issues. The detection of resistance associated genotypes in field samples using the described molecular assays provides rapid results compared to conventional phenotypic testing which requires culturing of fungal isolates. Further monitoring of resistance to critical Botryticidal chemistry is essential to improving current resistance management strategies.