Diseases of Aquatic Organisms | |
Detection of infectious salmon anaemia virus by real-time nucleic acid sequence based amplification | |
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
【 预 览 】
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RO201911300283056ZK.pdf | 115KB | download |