Allium white rot caused by the pathogenic fungus Sclerotium cepivorum is a devastating disease of garlic worldwide. In this study, we developed and evaluated a molecular assay for the detection of S. cepivorumin mycelia and infected garlic cloves, through PCR, using a specific primer set combined with a rapid boiling DNA extraction. The PCR products were amplified in all the samples infected with S. cepivorum, hut not in other samples including phylogenetically closely related fungi. This methodology is specific and has a shorter execution time compared to standard methods.