Development of a Real-time Polymerase Chain Reaction for the Detection of Fusarium oxysporum f. sp. basilici from Basil Seed and Roots

A real-time polymerase chain reaction (PCR) assay using a TaqMan probe was developed to detect the causal agent of wilt and crown rot of basil from infected plants and seed in Italy. The aim of the study was to diminish testing time, previously performed using nested-PCR, and to create the conditions for future automation. The sensitivity of the assay was shown to be similar to the detection limit of the available nested- PCR procedure. The advantages of real-time PCR system include halving of the testing time, as well as the ability to identify both internally and externally infected seed to the sensitivity of 1 pg of genomic DNA. The assay was able to detect the presence of the pathogen in infected seed up to a sensitivity of 24 (SD: ±10) CFU per 100 seed