Development and validation of a real‐time PCR assay for the detection of anguillid herpesvirus 1

Anguillid herpesvirus 1 (Ang HV 1) causes a haemorrhagic disease with increased mortality in wild and farmed European eel, Anguilla anguilla (L.) and Japanese eel Anguilla japonica , Temminck & Schlegel). Detection of Ang HV 1 is currently based on virus isolation in cell culture, antibody‐based typing assays or conventional PCR . We developed, optimized and concisely validated a diagnostic TaqMan probe based real‐time PCR assay for the detection of Ang HV 1. The primers and probe target Ang HV 1 open reading frame 57, encoding the capsid protease and scaffold protein. Compared to conventional PCR , the developed real‐time PCR is faster, less labour‐intensive and has a reduced risk of cross‐contamination. The real‐time PCR assay was shown to be analytically sensitive and specific and has a high repeatability, efficiency and r 2 ‐value. The diagnostic performance of the assay was determined by testing 10% w/v organ suspensions and virus cultures from wild and farmed European eels from the Netherlands by conventional and real‐time PCR . The developed real‐time PCR assay is a useful tool for the rapid and sensitive detection of Ang HV 1 in 10% w/v organ suspensions from wild and farmed European eels.