Detection of natural infection of infectious spleen and kidney necrosis virus in farmed tilapia by hydroxynapthol blue‐loop‐mediated isothermal amplification assay
Aims Infectious spleen and kidney necrosis virus (ISKNV) has recently been recognized as a causative agent of serious systemic disease in tilapia. Our objective was to establish a new colorimetric loop‐mediated isothermal amplification (LAMP) assay with pre‐addition of hydroxynapthol blue (blue‐LAMP...
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| Veröffentlicht in: | Journal of applied microbiology 2016-07, Vol.121 (1), p.55-67 |
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| creator | Suebsing, R. Pradeep, P.J. Jitrakorn, S. Sirithammajak, S. Kampeera, J. Turner, W.A. Saksmerprome, V. Withyachumnarnkul, B. Kiatpathomchai, W. |
| description | Aims
Infectious spleen and kidney necrosis virus (ISKNV) has recently been recognized as a causative agent of serious systemic disease in tilapia. Our objective was to establish a new colorimetric loop‐mediated isothermal amplification (LAMP) assay with pre‐addition of hydroxynapthol blue (blue‐LAMP) to investigate ISKNV transmission in tilapia.
Methods and Results
The blue‐LAMP, targeting a major capsid protein gene of ISKNV, was conducted at 65°C for 45 min, allowing unaided visual detection of the pathogen based on colour change without cross‐amplification of other known fish pathogens tested. Comparison of blue‐LAMP and PCR assays revealed a higher detection level for blue‐LAMP assay (41·33%) in a population of farmed tilapia infected with ISKNV. The investigation of ISKNV transmission pattern in farmed red tilapia using the blue‐LAMP revealed a possible matroclinical form. The presence of ISKNV in the gonad samples was confirmed by in situ LAMP assay. Positive signals only appeared in ovarian follicles, and not in oocytes. Moreover, tissue tropism assay revealed that the brain was the main target organ in both farmed red tilapia (40%) and Nile tilapia (20%).
Conclusions
The developed blue‐LAMP assay has the potential to be used as a viable tool for screening covert and natural infections of ISKNV in tilapia. The evidence of vertical transmission of ISKNV infection in tilapia indicates the seriousness of this disease and will require a close attention and collaboration between tilapia hatcheries and disease experts in order to find a solution.
Significance and Impact of the Study
The new blue‐LAMP assay is a time‐saving and economically viable detection tool, which allows unaided visual detection for ISKNV in tilapia, and it could be applicable for field applications. Evidence on the vertical transmission of ISKNV in farmed tilapia suggests a need for developing farm management practices to control the spread of virus in aquaculture industries. |
| doi_str_mv | 10.1111/jam.13165 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1808727614</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1799213678</sourcerecordid><originalsourceid>FETCH-LOGICAL-c5575-d27bf810ff611c03c2dd507d708bcc32c0beff7c26c5f1956e6dc731efef7d283</originalsourceid><addsrcrecordid>eNqFkc9uFSEUxonRtLXtwhcwJG7qYlpgLjB32bT1X2q6adcTBg65XBkYYUY7Ox_Bt_C9fBK597aamBjZHCA_Ps75PoReUHJKyzpbq_6U1lTwJ-iA1oJXTEj2dLtfVJxIto-e57wmhNaEiz20zyStGyIWB-jHJYygRxcDjhYHNU5JeeyC_XP5eJgyzoMHCFgFgz85E2DGAXSK2WX8xaUCuICtSj0YPDqvBqdwN-PVbFK8n4MaxlX0uPMT_Pz23cc4lFJYp8bywOU4riD15XvVD95Zp9W2BZWzmo_QM6t8huOHeoju3lzdXryrrm_evr84v64055JXhsnONpRYKyjVpNbMmGKAkaTptK6ZJh1YKzUTmlu65AKE0bKmYMFKw5r6EJ3sdIcUP0-Qx7Z3WYP3KkBxoKUNaSSTgi7-j8rlkpUI5Eb11V_oOk4plEE2VCOYbLbU6x21sTQnsO2QXK_S3FLSboJuS9DtNujCvnxQnLpi4W_yMdkCnO2Ar87D_G-l9sP5x53kL3icuE0</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1798627878</pqid></control><display><type>article</type><title>Detection of natural infection of infectious spleen and kidney necrosis virus in farmed tilapia by hydroxynapthol blue‐loop‐mediated isothermal amplification assay</title><source>MEDLINE</source><source>Oxford University Press Journals All Titles (1996-Current)</source><source>Wiley Online Library All Journals</source><creator>Suebsing, R. ; Pradeep, P.J. ; Jitrakorn, S. ; Sirithammajak, S. ; Kampeera, J. ; Turner, W.A. ; Saksmerprome, V. ; Withyachumnarnkul, B. ; Kiatpathomchai, W.</creator><creatorcontrib>Suebsing, R. ; Pradeep, P.J. ; Jitrakorn, S. ; Sirithammajak, S. ; Kampeera, J. ; Turner, W.A. ; Saksmerprome, V. ; Withyachumnarnkul, B. ; Kiatpathomchai, W.</creatorcontrib><description>Aims
Infectious spleen and kidney necrosis virus (ISKNV) has recently been recognized as a causative agent of serious systemic disease in tilapia. Our objective was to establish a new colorimetric loop‐mediated isothermal amplification (LAMP) assay with pre‐addition of hydroxynapthol blue (blue‐LAMP) to investigate ISKNV transmission in tilapia.
Methods and Results
The blue‐LAMP, targeting a major capsid protein gene of ISKNV, was conducted at 65°C for 45 min, allowing unaided visual detection of the pathogen based on colour change without cross‐amplification of other known fish pathogens tested. Comparison of blue‐LAMP and PCR assays revealed a higher detection level for blue‐LAMP assay (41·33%) in a population of farmed tilapia infected with ISKNV. The investigation of ISKNV transmission pattern in farmed red tilapia using the blue‐LAMP revealed a possible matroclinical form. The presence of ISKNV in the gonad samples was confirmed by in situ LAMP assay. Positive signals only appeared in ovarian follicles, and not in oocytes. Moreover, tissue tropism assay revealed that the brain was the main target organ in both farmed red tilapia (40%) and Nile tilapia (20%).
Conclusions
The developed blue‐LAMP assay has the potential to be used as a viable tool for screening covert and natural infections of ISKNV in tilapia. The evidence of vertical transmission of ISKNV infection in tilapia indicates the seriousness of this disease and will require a close attention and collaboration between tilapia hatcheries and disease experts in order to find a solution.
Significance and Impact of the Study
The new blue‐LAMP assay is a time‐saving and economically viable detection tool, which allows unaided visual detection for ISKNV in tilapia, and it could be applicable for field applications. Evidence on the vertical transmission of ISKNV in farmed tilapia suggests a need for developing farm management practices to control the spread of virus in aquaculture industries.</description><identifier>ISSN: 1364-5072</identifier><identifier>EISSN: 1365-2672</identifier><identifier>DOI: 10.1111/jam.13165</identifier><identifier>PMID: 27138064</identifier><identifier>CODEN: JAMIFK</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>Animals ; Aquaculture ; Aquaculture - methods ; Bacterial infections ; Bioassays ; Colorimetry - methods ; Fish Diseases - transmission ; Fish Diseases - virology ; hydroxynapthol blue ; infectious spleen and kidney necrosis virus ; in situ LAMP ; loop‐mediated isothermal amplification ; Microbiology ; Nucleic Acid Amplification Techniques ; Oreochromis niloticus ; Retroviridae Infections - diagnosis ; Retroviridae Infections - transmission ; Retroviridae Infections - veterinary ; Retroviridae Infections - virology ; Sensitivity and Specificity ; Tilapia ; Tilapia - virology ; tissue tropism ; Trager duck spleen necrosis virus - genetics ; Trager duck spleen necrosis virus - isolation & purification ; vertical transmission</subject><ispartof>Journal of applied microbiology, 2016-07, Vol.121 (1), p.55-67</ispartof><rights>2016 The Society for Applied Microbiology</rights><rights>2016 The Society for Applied Microbiology.</rights><rights>Copyright © 2016 The Society for Applied Microbiology</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5575-d27bf810ff611c03c2dd507d708bcc32c0beff7c26c5f1956e6dc731efef7d283</citedby><cites>FETCH-LOGICAL-c5575-d27bf810ff611c03c2dd507d708bcc32c0beff7c26c5f1956e6dc731efef7d283</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fjam.13165$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fjam.13165$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27138064$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Suebsing, R.</creatorcontrib><creatorcontrib>Pradeep, P.J.</creatorcontrib><creatorcontrib>Jitrakorn, S.</creatorcontrib><creatorcontrib>Sirithammajak, S.</creatorcontrib><creatorcontrib>Kampeera, J.</creatorcontrib><creatorcontrib>Turner, W.A.</creatorcontrib><creatorcontrib>Saksmerprome, V.</creatorcontrib><creatorcontrib>Withyachumnarnkul, B.</creatorcontrib><creatorcontrib>Kiatpathomchai, W.</creatorcontrib><title>Detection of natural infection of infectious spleen and kidney necrosis virus in farmed tilapia by hydroxynapthol blue‐loop‐mediated isothermal amplification assay</title><title>Journal of applied microbiology</title><addtitle>J Appl Microbiol</addtitle><description>Aims
Infectious spleen and kidney necrosis virus (ISKNV) has recently been recognized as a causative agent of serious systemic disease in tilapia. Our objective was to establish a new colorimetric loop‐mediated isothermal amplification (LAMP) assay with pre‐addition of hydroxynapthol blue (blue‐LAMP) to investigate ISKNV transmission in tilapia.
Methods and Results
The blue‐LAMP, targeting a major capsid protein gene of ISKNV, was conducted at 65°C for 45 min, allowing unaided visual detection of the pathogen based on colour change without cross‐amplification of other known fish pathogens tested. Comparison of blue‐LAMP and PCR assays revealed a higher detection level for blue‐LAMP assay (41·33%) in a population of farmed tilapia infected with ISKNV. The investigation of ISKNV transmission pattern in farmed red tilapia using the blue‐LAMP revealed a possible matroclinical form. The presence of ISKNV in the gonad samples was confirmed by in situ LAMP assay. Positive signals only appeared in ovarian follicles, and not in oocytes. Moreover, tissue tropism assay revealed that the brain was the main target organ in both farmed red tilapia (40%) and Nile tilapia (20%).
Conclusions
The developed blue‐LAMP assay has the potential to be used as a viable tool for screening covert and natural infections of ISKNV in tilapia. The evidence of vertical transmission of ISKNV infection in tilapia indicates the seriousness of this disease and will require a close attention and collaboration between tilapia hatcheries and disease experts in order to find a solution.
Significance and Impact of the Study
The new blue‐LAMP assay is a time‐saving and economically viable detection tool, which allows unaided visual detection for ISKNV in tilapia, and it could be applicable for field applications. Evidence on the vertical transmission of ISKNV in farmed tilapia suggests a need for developing farm management practices to control the spread of virus in aquaculture industries.</description><subject>Animals</subject><subject>Aquaculture</subject><subject>Aquaculture - methods</subject><subject>Bacterial infections</subject><subject>Bioassays</subject><subject>Colorimetry - methods</subject><subject>Fish Diseases - transmission</subject><subject>Fish Diseases - virology</subject><subject>hydroxynapthol blue</subject><subject>infectious spleen and kidney necrosis virus</subject><subject>in situ LAMP</subject><subject>loop‐mediated isothermal amplification</subject><subject>Microbiology</subject><subject>Nucleic Acid Amplification Techniques</subject><subject>Oreochromis niloticus</subject><subject>Retroviridae Infections - diagnosis</subject><subject>Retroviridae Infections - transmission</subject><subject>Retroviridae Infections - veterinary</subject><subject>Retroviridae Infections - virology</subject><subject>Sensitivity and Specificity</subject><subject>Tilapia</subject><subject>Tilapia - virology</subject><subject>tissue tropism</subject><subject>Trager duck spleen necrosis virus - genetics</subject><subject>Trager duck spleen necrosis virus - isolation & purification</subject><subject>vertical transmission</subject><issn>1364-5072</issn><issn>1365-2672</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc9uFSEUxonRtLXtwhcwJG7qYlpgLjB32bT1X2q6adcTBg65XBkYYUY7Ox_Bt_C9fBK597aamBjZHCA_Ps75PoReUHJKyzpbq_6U1lTwJ-iA1oJXTEj2dLtfVJxIto-e57wmhNaEiz20zyStGyIWB-jHJYygRxcDjhYHNU5JeeyC_XP5eJgyzoMHCFgFgz85E2DGAXSK2WX8xaUCuICtSj0YPDqvBqdwN-PVbFK8n4MaxlX0uPMT_Pz23cc4lFJYp8bywOU4riD15XvVD95Zp9W2BZWzmo_QM6t8huOHeoju3lzdXryrrm_evr84v64055JXhsnONpRYKyjVpNbMmGKAkaTptK6ZJh1YKzUTmlu65AKE0bKmYMFKw5r6EJ3sdIcUP0-Qx7Z3WYP3KkBxoKUNaSSTgi7-j8rlkpUI5Eb11V_oOk4plEE2VCOYbLbU6x21sTQnsO2QXK_S3FLSboJuS9DtNujCvnxQnLpi4W_yMdkCnO2Ar87D_G-l9sP5x53kL3icuE0</recordid><startdate>201607</startdate><enddate>201607</enddate><creator>Suebsing, R.</creator><creator>Pradeep, P.J.</creator><creator>Jitrakorn, S.</creator><creator>Sirithammajak, S.</creator><creator>Kampeera, J.</creator><creator>Turner, W.A.</creator><creator>Saksmerprome, V.</creator><creator>Withyachumnarnkul, B.</creator><creator>Kiatpathomchai, W.</creator><general>Oxford University Press</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>7TM</scope><scope>7U7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>7U9</scope><scope>H94</scope></search><sort><creationdate>201607</creationdate><title>Detection of natural infection of infectious spleen and kidney necrosis virus in farmed tilapia by hydroxynapthol blue‐loop‐mediated isothermal amplification assay</title><author>Suebsing, R. ; Pradeep, P.J. ; Jitrakorn, S. ; Sirithammajak, S. ; Kampeera, J. ; Turner, W.A. ; Saksmerprome, V. ; Withyachumnarnkul, B. ; Kiatpathomchai, W.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5575-d27bf810ff611c03c2dd507d708bcc32c0beff7c26c5f1956e6dc731efef7d283</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Animals</topic><topic>Aquaculture</topic><topic>Aquaculture - methods</topic><topic>Bacterial infections</topic><topic>Bioassays</topic><topic>Colorimetry - methods</topic><topic>Fish Diseases - transmission</topic><topic>Fish Diseases - virology</topic><topic>hydroxynapthol blue</topic><topic>infectious spleen and kidney necrosis virus</topic><topic>in situ LAMP</topic><topic>loop‐mediated isothermal amplification</topic><topic>Microbiology</topic><topic>Nucleic Acid Amplification Techniques</topic><topic>Oreochromis niloticus</topic><topic>Retroviridae Infections - diagnosis</topic><topic>Retroviridae Infections - transmission</topic><topic>Retroviridae Infections - veterinary</topic><topic>Retroviridae Infections - virology</topic><topic>Sensitivity and Specificity</topic><topic>Tilapia</topic><topic>Tilapia - virology</topic><topic>tissue tropism</topic><topic>Trager duck spleen necrosis virus - genetics</topic><topic>Trager duck spleen necrosis virus - isolation & purification</topic><topic>vertical transmission</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Suebsing, R.</creatorcontrib><creatorcontrib>Pradeep, P.J.</creatorcontrib><creatorcontrib>Jitrakorn, S.</creatorcontrib><creatorcontrib>Sirithammajak, S.</creatorcontrib><creatorcontrib>Kampeera, J.</creatorcontrib><creatorcontrib>Turner, W.A.</creatorcontrib><creatorcontrib>Saksmerprome, V.</creatorcontrib><creatorcontrib>Withyachumnarnkul, B.</creatorcontrib><creatorcontrib>Kiatpathomchai, W.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><jtitle>Journal of applied microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Suebsing, R.</au><au>Pradeep, P.J.</au><au>Jitrakorn, S.</au><au>Sirithammajak, S.</au><au>Kampeera, J.</au><au>Turner, W.A.</au><au>Saksmerprome, V.</au><au>Withyachumnarnkul, B.</au><au>Kiatpathomchai, W.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of natural infection of infectious spleen and kidney necrosis virus in farmed tilapia by hydroxynapthol blue‐loop‐mediated isothermal amplification assay</atitle><jtitle>Journal of applied microbiology</jtitle><addtitle>J Appl Microbiol</addtitle><date>2016-07</date><risdate>2016</risdate><volume>121</volume><issue>1</issue><spage>55</spage><epage>67</epage><pages>55-67</pages><issn>1364-5072</issn><eissn>1365-2672</eissn><coden>JAMIFK</coden><abstract>Aims
Infectious spleen and kidney necrosis virus (ISKNV) has recently been recognized as a causative agent of serious systemic disease in tilapia. Our objective was to establish a new colorimetric loop‐mediated isothermal amplification (LAMP) assay with pre‐addition of hydroxynapthol blue (blue‐LAMP) to investigate ISKNV transmission in tilapia.
Methods and Results
The blue‐LAMP, targeting a major capsid protein gene of ISKNV, was conducted at 65°C for 45 min, allowing unaided visual detection of the pathogen based on colour change without cross‐amplification of other known fish pathogens tested. Comparison of blue‐LAMP and PCR assays revealed a higher detection level for blue‐LAMP assay (41·33%) in a population of farmed tilapia infected with ISKNV. The investigation of ISKNV transmission pattern in farmed red tilapia using the blue‐LAMP revealed a possible matroclinical form. The presence of ISKNV in the gonad samples was confirmed by in situ LAMP assay. Positive signals only appeared in ovarian follicles, and not in oocytes. Moreover, tissue tropism assay revealed that the brain was the main target organ in both farmed red tilapia (40%) and Nile tilapia (20%).
Conclusions
The developed blue‐LAMP assay has the potential to be used as a viable tool for screening covert and natural infections of ISKNV in tilapia. The evidence of vertical transmission of ISKNV infection in tilapia indicates the seriousness of this disease and will require a close attention and collaboration between tilapia hatcheries and disease experts in order to find a solution.
Significance and Impact of the Study
The new blue‐LAMP assay is a time‐saving and economically viable detection tool, which allows unaided visual detection for ISKNV in tilapia, and it could be applicable for field applications. Evidence on the vertical transmission of ISKNV in farmed tilapia suggests a need for developing farm management practices to control the spread of virus in aquaculture industries.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>27138064</pmid><doi>10.1111/jam.13165</doi><tpages>13</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Aquaculture Aquaculture - methods Bacterial infections Bioassays Colorimetry - methods Fish Diseases - transmission Fish Diseases - virology hydroxynapthol blue infectious spleen and kidney necrosis virus in situ LAMP loop‐mediated isothermal amplification Microbiology Nucleic Acid Amplification Techniques Oreochromis niloticus Retroviridae Infections - diagnosis Retroviridae Infections - transmission Retroviridae Infections - veterinary Retroviridae Infections - virology Sensitivity and Specificity Tilapia Tilapia - virology tissue tropism Trager duck spleen necrosis virus - genetics Trager duck spleen necrosis virus - isolation & purification vertical transmission |
| title | Detection of natural infection of infectious spleen and kidney necrosis virus in farmed tilapia by hydroxynapthol blue‐loop‐mediated isothermal amplification assay |
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