Detection of Edwardsiella ictaluri and Citrobacter freundii causing disease on Catfish ( Pangasianodon hypophthalmus ) by Multiplex PCR

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Abstract Intensive farming practices of striped catfish ( Pangasianodon hypophthalmus ) in Vietnam, for example, high stocking densities and accumulated waste, create conditions conducive to co-infection by Edwardsiella ictaluri and Citrobacter freundii , resulting in severe diseases like Bacillary Necrosis of Pangasius (BNP) and hemorrhagic enteritis. To establish a foundation for the simultaneous detection of these two pathogens, a multiplex PCR assay was developed targeting the species-specific t3ss gene of E. ictaluri and the tpl gene of C. freundii , generating amplicons of 208 bp and 508 bp, respectively. The assay successfully amplified both targets with high specificity and no cross-reactivity, demonstrating its potential as a rapid and cost-effective molecular tool for routine laboratory diagnosis and disease surveillance in striped catfish aquaculture.

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