Detection of infectious salmon anaemia virus by real-time nucleic acid sequence based amplification
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 beac...
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| Veröffentlicht in: | Diseases of aquatic organisms 2006-10, Vol.72 (2), p.107-113 |
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| container_title | Diseases of aquatic organisms |
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| creator | STARKEY, William G SMAIL, David A BLEIE, Hogne MUIR, K. Fiona IRELAND, Jacqueline H RICHARDS, Randolph H |
| description | 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 degrees 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 100x more sensitive than conventional RT-PCR. These results suggest that real-time NASBA may represent a useful diagnostic procedure for ISAV. |
| doi_str_mv | 10.3354/dao072107 |
| format | Article |
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Fiona ; IRELAND, Jacqueline H ; RICHARDS, Randolph H</creator><creatorcontrib>STARKEY, William G ; SMAIL, David A ; BLEIE, Hogne ; MUIR, K. Fiona ; IRELAND, Jacqueline H ; RICHARDS, Randolph H</creatorcontrib><description>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 degrees 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. 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Fiona</creatorcontrib><creatorcontrib>IRELAND, Jacqueline H</creatorcontrib><creatorcontrib>RICHARDS, Randolph H</creatorcontrib><title>Detection of infectious salmon anaemia virus by real-time nucleic acid sequence based amplification</title><title>Diseases of aquatic organisms</title><addtitle>Dis Aquat Organ</addtitle><description>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 degrees 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 100x more sensitive than conventional RT-PCR. These results suggest that real-time NASBA may represent a useful diagnostic procedure for ISAV.</description><subject>Animal aquaculture</subject><subject>Animal productions</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>DNA Primers - chemistry</subject><subject>Fish Diseases - diagnosis</subject><subject>Fish Diseases - virology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Infectious salmon anemia virus</subject><subject>Isavirus - genetics</subject><subject>Isavirus - isolation & purification</subject><subject>Kidney - virology</subject><subject>Orthomyxoviridae Infections - diagnosis</subject><subject>Orthomyxoviridae Infections - veterinary</subject><subject>Orthomyxoviridae Infections - virology</subject><subject>Pisciculture</subject><subject>Reverse Transcriptase Polymerase Chain Reaction - veterinary</subject><subject>RNA, Viral - isolation & purification</subject><subject>Salmo salar - virology</subject><subject>Salmonidae</subject><subject>Self-Sustained Sequence Replication - methods</subject><subject>Self-Sustained Sequence Replication - veterinary</subject><subject>Sensitivity and Specificity</subject><subject>Time Factors</subject><subject>Vertebrate aquaculture</subject><issn>0177-5103</issn><issn>1616-1580</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0EtLxDAUBeAgio6jC_-AZKPgonpv0jTNUnyD4EbX5Ta9hUgfY9MR5t9btThLVyGHj0NyhDhBuNTapFcV9WAVgt0RC8wwS9DksCsWgNYmBkEfiMMY3wFQOYP74gAtpoBaL4S_5ZH9GPpO9rUMXf1zWUcZqWmnkDriNpD8DMMUlhs5MDXJGFqW3do3HLwkHyoZ-WPNnWdZUuRKUrtqQh08fTcfib2amsjH87kUb_d3rzePyfPLw9PN9XPitXVjUoGzgM4pysvakyPDxLqsakM6Z841pCWm1kyRQ6UzqKjUVeoYvFd5qfVSnP_2roZ-ek0cizZEz01DHU9fKrJcgYIs_Rei09YojRO8-IV-6GMcuC5WQ2hp2BQIxff0xd_0kz2dS9dly9VWzltP4GwGFD019UCdD3HrcpVZB0Z_AQktjJA</recordid><startdate>20061017</startdate><enddate>20061017</enddate><creator>STARKEY, William G</creator><creator>SMAIL, David A</creator><creator>BLEIE, Hogne</creator><creator>MUIR, K. 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Fiona</au><au>IRELAND, Jacqueline H</au><au>RICHARDS, Randolph H</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of infectious salmon anaemia virus by real-time nucleic acid sequence based amplification</atitle><jtitle>Diseases of aquatic organisms</jtitle><addtitle>Dis Aquat Organ</addtitle><date>2006-10-17</date><risdate>2006</risdate><volume>72</volume><issue>2</issue><spage>107</spage><epage>113</epage><pages>107-113</pages><issn>0177-5103</issn><eissn>1616-1580</eissn><coden>DAOREO</coden><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. 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| ispartof | Diseases of aquatic organisms, 2006-10, Vol.72 (2), p.107-113 |
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| source | MEDLINE; Inter-Research; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Animal aquaculture Animal productions Animals Biological and medical sciences DNA Primers - chemistry Fish Diseases - diagnosis Fish Diseases - virology Fundamental and applied biological sciences. Psychology Infectious salmon anemia virus Isavirus - genetics Isavirus - isolation & purification Kidney - virology Orthomyxoviridae Infections - diagnosis Orthomyxoviridae Infections - veterinary Orthomyxoviridae Infections - virology Pisciculture Reverse Transcriptase Polymerase Chain Reaction - veterinary RNA, Viral - isolation & purification Salmo salar - virology Salmonidae Self-Sustained Sequence Replication - methods Self-Sustained Sequence Replication - veterinary Sensitivity and Specificity Time Factors Vertebrate aquaculture |
| title | Detection of infectious salmon anaemia virus by real-time nucleic acid sequence based amplification |
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