Detection of shrimp Taura syndrome virus by loop-mediated isothermal amplification using a designed portable multi-channel turbidimeter
In this study, a portable turbidimetric end-point detection method was devised and tested for the detection of Taura syndrome virus (TSV) using spectroscopic measurement of a loop-mediated isothermal amplification (LAMP) by-product: magnesium pyrophosphate (Mg 2P 2O 7). The device incorporated a hea...
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| Veröffentlicht in: | Journal of virological methods 2011-08, Vol.175 (2), p.141-148 |
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| container_title | Journal of virological methods |
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| creator | Sappat, Assawapong Jaroenram, Wansadaj Puthawibool, Teeranart Lomas, Tanom Tuantranont, Adisorn Kiatpathomchai, Wansika |
| description | In this study, a portable turbidimetric end-point detection method was devised and tested for the detection of Taura syndrome virus (TSV) using spectroscopic measurement of a loop-mediated isothermal amplification (LAMP) by-product: magnesium pyrophosphate (Mg
2P
2O
7). The device incorporated a heating block that maintained an optimal temperature of 63
°C for the duration of the RT-LAMP reaction. Turbidity of the RT-LAMP by-product was measured when light from a light-emitting diode (LED) passed through the tube to reach a light dependent resistance (LDR) detector. Results revealed that turbidity measurement of the RT-LAMP reactions using this device provided the same detection sensitivity as the agarose gel electrophoresis detection of RT-LAMP and nested RT-PCR (IQ2000™) products. Cross reactions with other shrimp viruses were not found, indicating that the RT-LAMP-turbidity measurement was highly specific to TSV. The combination of 10
min for rapid RNA preparation with 30
min for RT-LAMP amplification followed by turbidity measurement resulted in a total assay time of less than 1
h compared to 4–8
h for the nested RT-PCR method. RT-LAMP plus turbidity measurement constitutes a platform for the development of more rapid and user-friendly detection of TSV in the field. |
| doi_str_mv | 10.1016/j.jviromet.2011.05.013 |
| format | Article |
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2P
2O
7). The device incorporated a heating block that maintained an optimal temperature of 63
°C for the duration of the RT-LAMP reaction. Turbidity of the RT-LAMP by-product was measured when light from a light-emitting diode (LED) passed through the tube to reach a light dependent resistance (LDR) detector. Results revealed that turbidity measurement of the RT-LAMP reactions using this device provided the same detection sensitivity as the agarose gel electrophoresis detection of RT-LAMP and nested RT-PCR (IQ2000™) products. Cross reactions with other shrimp viruses were not found, indicating that the RT-LAMP-turbidity measurement was highly specific to TSV. The combination of 10
min for rapid RNA preparation with 30
min for RT-LAMP amplification followed by turbidity measurement resulted in a total assay time of less than 1
h compared to 4–8
h for the nested RT-PCR method. RT-LAMP plus turbidity measurement constitutes a platform for the development of more rapid and user-friendly detection of TSV in the field.</description><identifier>ISSN: 0166-0934</identifier><identifier>EISSN: 1879-0984</identifier><identifier>DOI: 10.1016/j.jviromet.2011.05.013</identifier><identifier>PMID: 21619895</identifier><identifier>CODEN: JVMEDH</identifier><language>eng</language><publisher>Kidlington: Elsevier B.V</publisher><subject>Animals ; Base Sequence ; Biological and medical sciences ; Dicistroviridae - genetics ; Dicistroviridae - isolation & purification ; DNA Primers - genetics ; Fundamental and applied biological sciences. Psychology ; Loop-mediated isothermal amplification ; Microbiology ; Molecular Sequence Data ; Nephelometry and Turbidimetry - methods ; Nucleic Acid Amplification Techniques - methods ; PCR ; Penaeidae - virology ; Sensitivity and Specificity ; Taura syndrome virus ; Techniques used in virology ; Time Factors ; TSV ; Turbidity ; Virology ; Virology - methods</subject><ispartof>Journal of virological methods, 2011-08, Vol.175 (2), p.141-148</ispartof><rights>2011 Elsevier B.V.</rights><rights>2015 INIST-CNRS</rights><rights>Copyright © 2011 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c495t-f8661481ead1252a000e97bd022abb307c7190f48ec59f10724f5207815c58fa3</citedby><cites>FETCH-LOGICAL-c495t-f8661481ead1252a000e97bd022abb307c7190f48ec59f10724f5207815c58fa3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.jviromet.2011.05.013$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=24280705$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21619895$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sappat, Assawapong</creatorcontrib><creatorcontrib>Jaroenram, Wansadaj</creatorcontrib><creatorcontrib>Puthawibool, Teeranart</creatorcontrib><creatorcontrib>Lomas, Tanom</creatorcontrib><creatorcontrib>Tuantranont, Adisorn</creatorcontrib><creatorcontrib>Kiatpathomchai, Wansika</creatorcontrib><title>Detection of shrimp Taura syndrome virus by loop-mediated isothermal amplification using a designed portable multi-channel turbidimeter</title><title>Journal of virological methods</title><addtitle>J Virol Methods</addtitle><description>In this study, a portable turbidimetric end-point detection method was devised and tested for the detection of Taura syndrome virus (TSV) using spectroscopic measurement of a loop-mediated isothermal amplification (LAMP) by-product: magnesium pyrophosphate (Mg
2P
2O
7). The device incorporated a heating block that maintained an optimal temperature of 63
°C for the duration of the RT-LAMP reaction. Turbidity of the RT-LAMP by-product was measured when light from a light-emitting diode (LED) passed through the tube to reach a light dependent resistance (LDR) detector. Results revealed that turbidity measurement of the RT-LAMP reactions using this device provided the same detection sensitivity as the agarose gel electrophoresis detection of RT-LAMP and nested RT-PCR (IQ2000™) products. Cross reactions with other shrimp viruses were not found, indicating that the RT-LAMP-turbidity measurement was highly specific to TSV. The combination of 10
min for rapid RNA preparation with 30
min for RT-LAMP amplification followed by turbidity measurement resulted in a total assay time of less than 1
h compared to 4–8
h for the nested RT-PCR method. RT-LAMP plus turbidity measurement constitutes a platform for the development of more rapid and user-friendly detection of TSV in the field.</description><subject>Animals</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Dicistroviridae - genetics</subject><subject>Dicistroviridae - isolation & purification</subject><subject>DNA Primers - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Loop-mediated isothermal amplification</subject><subject>Microbiology</subject><subject>Molecular Sequence Data</subject><subject>Nephelometry and Turbidimetry - methods</subject><subject>Nucleic Acid Amplification Techniques - methods</subject><subject>PCR</subject><subject>Penaeidae - virology</subject><subject>Sensitivity and Specificity</subject><subject>Taura syndrome virus</subject><subject>Techniques used in virology</subject><subject>Time Factors</subject><subject>TSV</subject><subject>Turbidity</subject><subject>Virology</subject><subject>Virology - methods</subject><issn>0166-0934</issn><issn>1879-0984</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc9u3CAQh1HVqtmkfYWIS9WTt4DBhlur9E8qReolPSOMhywrbFzAkfYJ-tplu5v2mBNz-GaG-X0IXVOypYR2H_bb_aNPcYKyZYTSLRFbQtsXaENlrxqiJH-JNhXsat3yC3SZ854QIvq2fY0uGO2okkps0O_PUMAWH2ccHc675KcF35s1GZwP83jcgOuiNePhgEOMSzPB6E2BEfscyw7SZAI20xK889b8HbRmPz9gg0fI_mGu5BJTMUMAPK2h-MbuzDxDwGVNgx99vQHSG_TKmZDh7fm9Qj-_frm_uW3ufnz7fvPprrFcidI42XWUSwpmpEwwU08C1Q8jYcwMQ0t621NFHJdghXKU9Iw7wUgvqbBCOtNeofenuUuKv1bIRU8-WwjBzBDXrBXhXBIh2LOk7FuuKttWsjuRNsWcEzi91BhNOmhK9NGW3usnW_poSxOhq63aeH1esQ411n9tT3oq8O4MmGxNcMnM1uf_HGeS9OTIfTxxUKN79JB0th5mW1WlaleP0T_3lz-C6rkX</recordid><startdate>20110801</startdate><enddate>20110801</enddate><creator>Sappat, Assawapong</creator><creator>Jaroenram, Wansadaj</creator><creator>Puthawibool, Teeranart</creator><creator>Lomas, Tanom</creator><creator>Tuantranont, Adisorn</creator><creator>Kiatpathomchai, Wansika</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><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>7X8</scope><scope>7TN</scope><scope>7U9</scope><scope>F1W</scope><scope>H94</scope><scope>H95</scope><scope>L.G</scope></search><sort><creationdate>20110801</creationdate><title>Detection of shrimp Taura syndrome virus by loop-mediated isothermal amplification using a designed portable multi-channel turbidimeter</title><author>Sappat, Assawapong ; Jaroenram, Wansadaj ; Puthawibool, Teeranart ; Lomas, Tanom ; Tuantranont, Adisorn ; Kiatpathomchai, Wansika</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c495t-f8661481ead1252a000e97bd022abb307c7190f48ec59f10724f5207815c58fa3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Animals</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Dicistroviridae - genetics</topic><topic>Dicistroviridae - isolation & purification</topic><topic>DNA Primers - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Loop-mediated isothermal amplification</topic><topic>Microbiology</topic><topic>Molecular Sequence Data</topic><topic>Nephelometry and Turbidimetry - methods</topic><topic>Nucleic Acid Amplification Techniques - methods</topic><topic>PCR</topic><topic>Penaeidae - virology</topic><topic>Sensitivity and Specificity</topic><topic>Taura syndrome virus</topic><topic>Techniques used in virology</topic><topic>Time Factors</topic><topic>TSV</topic><topic>Turbidity</topic><topic>Virology</topic><topic>Virology - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sappat, Assawapong</creatorcontrib><creatorcontrib>Jaroenram, Wansadaj</creatorcontrib><creatorcontrib>Puthawibool, Teeranart</creatorcontrib><creatorcontrib>Lomas, Tanom</creatorcontrib><creatorcontrib>Tuantranont, Adisorn</creatorcontrib><creatorcontrib>Kiatpathomchai, Wansika</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Oceanic Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><jtitle>Journal of virological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sappat, Assawapong</au><au>Jaroenram, Wansadaj</au><au>Puthawibool, Teeranart</au><au>Lomas, Tanom</au><au>Tuantranont, Adisorn</au><au>Kiatpathomchai, Wansika</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of shrimp Taura syndrome virus by loop-mediated isothermal amplification using a designed portable multi-channel turbidimeter</atitle><jtitle>Journal of virological methods</jtitle><addtitle>J Virol Methods</addtitle><date>2011-08-01</date><risdate>2011</risdate><volume>175</volume><issue>2</issue><spage>141</spage><epage>148</epage><pages>141-148</pages><issn>0166-0934</issn><eissn>1879-0984</eissn><coden>JVMEDH</coden><abstract>In this study, a portable turbidimetric end-point detection method was devised and tested for the detection of Taura syndrome virus (TSV) using spectroscopic measurement of a loop-mediated isothermal amplification (LAMP) by-product: magnesium pyrophosphate (Mg
2P
2O
7). The device incorporated a heating block that maintained an optimal temperature of 63
°C for the duration of the RT-LAMP reaction. Turbidity of the RT-LAMP by-product was measured when light from a light-emitting diode (LED) passed through the tube to reach a light dependent resistance (LDR) detector. Results revealed that turbidity measurement of the RT-LAMP reactions using this device provided the same detection sensitivity as the agarose gel electrophoresis detection of RT-LAMP and nested RT-PCR (IQ2000™) products. Cross reactions with other shrimp viruses were not found, indicating that the RT-LAMP-turbidity measurement was highly specific to TSV. The combination of 10
min for rapid RNA preparation with 30
min for RT-LAMP amplification followed by turbidity measurement resulted in a total assay time of less than 1
h compared to 4–8
h for the nested RT-PCR method. RT-LAMP plus turbidity measurement constitutes a platform for the development of more rapid and user-friendly detection of TSV in the field.</abstract><cop>Kidlington</cop><pub>Elsevier B.V</pub><pmid>21619895</pmid><doi>10.1016/j.jviromet.2011.05.013</doi><tpages>8</tpages></addata></record> |
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| ispartof | Journal of virological methods, 2011-08, Vol.175 (2), p.141-148 |
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| language | eng |
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| subjects | Animals Base Sequence Biological and medical sciences Dicistroviridae - genetics Dicistroviridae - isolation & purification DNA Primers - genetics Fundamental and applied biological sciences. Psychology Loop-mediated isothermal amplification Microbiology Molecular Sequence Data Nephelometry and Turbidimetry - methods Nucleic Acid Amplification Techniques - methods PCR Penaeidae - virology Sensitivity and Specificity Taura syndrome virus Techniques used in virology Time Factors TSV Turbidity Virology Virology - methods |
| title | Detection of shrimp Taura syndrome virus by loop-mediated isothermal amplification using a designed portable multi-channel turbidimeter |
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