Real-Time PCR Assays for Rapid Identification of Common Aphanomyces astaci Genotypes

The oomycete Aphanomyces astaci is the etiologic agent of crayfish plague, a disease that has seriously impacted populations European native species. introduction non-indigenous North American origin and their wide distribution across Europe have largely contributed to spread plague in areas populated by indigenous crayfish. Tracking A. genotypes may thus be useful tool for investigating natural history its range, as well sources pathways pathogen. In this study, we describe development real-time PCR TaqMan assays aiming distinguish five genotype groups (A–E) previously defined distinct RAPD patterns. method was evaluated using DNA extracts from pure cultures representing known groups, -positive clinical samples collected mostly during outbreaks recently occurred Central Italy Czechia. do not cross-react with each other, those targeting A, B, D, E seem sufficiently specific pathogen infected invaded (particularly Europe). unusual “SSR-Up” documented Czechia chronically Pontastacus leptodactylus Danube detected group B PCR. assay originally developed detect C (one yet outbreaks) showed cross-reactivity fennicus ; “rust1” described United States Faxonius rusticus well. Analyses additional markers (such sequencing nuclear internal transcribed spacer or mitochondrial ribosomal subunits) complement such cases when PCR-based genotyping conclusive. Despite some limitations, robust fast common Europe, both latent infections.