Servizio Comunitario di Informazione in materia di Ricerca e Sviluppo - CORDIS

Development of real time RT-PCR test for detecting SAV

A real-time RT-PCR test has been developed within the project by Partner 1. This test was developed to provide increased sensitivity for detecting SAV RNA because the conventional RT-PCR test developed by Partner 2 did not appear to have maximal sensitivity. The real time test was also developed to allow quantitative or semi-quantitative estimates of the amounts of virus RNA in experimental and clinical samples, thereby providing useful information about pathogenesis and virus pathogenicity.
The primers used for the real-time test were selected following comparison of partial gene sequences of 18 SAV isolates sourced from clinical outbreaks of PD and SD in Ireland, UK, France and Norway, with conserved regions being selected to ensure maximal chance of detecting all SAVs. The test was optimised and validated using RNA extracted from dilutions of cell culture-grown virus pools and serum samples of known infectious virus titre. The developed test was capable of detecting equal to or less than 1.5 TCID 50. It was more sensitive than the cell culture growth method for detecting infectious virus in serum. In addition it was successful in detecting SAV RNA in the heart tissue samples from the majority of SPDV experimentally infected fish from 7 to 70 days post-infection.

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Queen's University of Belfast
Veterinary Sciences Division, Stormont
BT4 3SD Belfast
United Kingdom