The Development of Tools for Identification and Diagnosis
Mortality due to LCD was initially determined from anecdotal observations or inference from clinical signs. To improve estimations, a set of species-specific PCR primers was developed in order to allow correct diagnosis, fungal identification, and environmental prospection of the species involved in LCD outbreaks (Pie et al.
2011; Guerra et al. 2013). All primers sets were based on sequences of rDNA internal transcribed spacer (ITS) regions. PCR products generated from extracts of samples (environment, colonies, crab tissue) differed between the two species and remained negative if the respective DNA targets were not present, allowing unambiguous identification of the agent. Amplicons measured 450 or 396 bp, corresponding to the expected product sizes of E. cancerae (strain CBS 120420) and F. brasiliensis (strain CBS 119710), respectively.11.5
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