All authors read and approved the final manuscript.”
“Background Botrytis cinerea is a pathogen ascomycete, which causes gray mold on a large number of economically important agricultural and horticultural crops [1–4]. This ubiquitous fungal pathogen is present often as latent infection. Latency is generally defined as the period between infection and the appearance AZD9291 of visible symptoms and can in the case of B. cinerea be long and variable [5–8]. Consequently, an apparently healthy fruit can deteriorate suddenly due to the development of this latent infection [9, 10]. Many synthetic fungicides are used as the principal mean of controlling

this important postharvest disease [11]. However, the growing public concern over the health and environmental hazards associated with fungicide use in orchards, the development of fungicide resistant strains of B. cinerea [12], and the deregistration of some of the most effective fungicides [13], have generated a great interest in the development of alternative methods to control the postharvest disease caused by this fungal pathogen. To prevent the indiscriminate use of fungicides, a sensitive and reliable method to early determination of the fungus in fruit tissues becomes crucial. The ability to detect latent infections in fruit

tissues should prove useful not only for early disease management but also for identifying infected fruit in postharvest. In addition, the quantification of the pathogen is necessary for the application of alternative methods of control, such as biological control using antagonist microorganisms because the success Ureohydrolase AR-13324 datasheet of this method depend of the ratio antagonist/pathogen [14]. The detection of fungus in fruit includes classical methods such as isolation on selective media, which is useful but subject to limitations [15] due to many pathogens can be masked by overgrowth of faster growing fungi. Other methods, such as quantitative real-time polymerase chain reaction (Q-PCR), or reverse transcription

polymerase chain reaction (RT-PCR) represent new tools for the detection of the pathogens by determination of their DNA/RNA [16–25]. Unfortunately these methods are expensive and not easy to perform routinely, because they require highly qualified personnel and need sophisticated instrumentation [26, 27]. In addition, to methods mentioned previously, some direct enzyme-linked immunosorbent assays (ELISAs) using microtiter plates have been developed for the detection of B. cinerea in pear steam, grape juice, and plants [28–32], but at present has not been reported any validated method based in an indirect competitive immunoassay for detection and quantification of the mentioned fungus in tissues of fruits. The aim of this study was the development and corroboration of a sensitive and specific ELISA for B.