Starkey W, Smail DA, Bleie H, Muir K, Ireland J & Richards R (2006) Detection of infectious salmon anaemia virus by real-time nucleic acid sequence based amplification. Diseases of Aquatic Organisms, 72 (2), pp. 107-113. https://doi.org/10.3354/dao072107
We have developed a real-time nucleic acid sequence based amplification (NASBA) procedure for detection of infectious salmon anaemia virus (ISAV). Primers were designed to target a 124 nucleotide region of ISAV genome segment 8. Amplification products were detected in real-time with a molecular beacon (carboxyfluorescin [FAM]-labelled and methyl-red quenched) that recognised an internal region of the target amplicon. Amplification and detection were performed at 41°C for 90 min in a Corbett Research Rotorgene. The real-time NASBA assay was compared to a conventional RT-PCR for ISAV detection. From a panel of 45 clinical samples, both assays detected ISAV in the same 19 samples. Based on the detection of a synthetic RNA target, the real-time NASBA procedure was approximately 100× more sensitive than conventional RT-PCR. These results suggest that real-time NASBA may represent a useful diagnostic procedure for ISAV.
Infectious salmon anaemia virus; Orthomyxovirus; NASBA; Diagnostics; Fish; Nucleic acid amplification
Diseases of Aquatic Organisms: Volume 72, Issue 2