Detection of infectious salmon anaemia virus by real-time nucleic acid sequence based amplification

Diseases of Aquatic Organisms

David A. Smail¹  William G. Starkey¹  K. Fiona Muir¹  Hogne Bleie¹  Jacqueline H. Ireland¹  Randolph H. Richards¹ 

关键词: Infectious salmon anaemia virus;    Orthomyxovirus;    NASBA;    Diagnostics;    Fish;    Nucleic acid amplification;   

DOI  :  10.3354/dao072107

学科分类：生物科学（综合）

来源: Inter-Research

【 摘 要 】

ABSTRACT: 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.

【 授权许可】

Unknown