Rapid discriminative detection of dengue viruses via loop mediated isothermal amplification
Dengue virus (DENV) is one of the life-threatening viruses to the human. In this study, we have designed specific novel primers for rapid discriminative detection of DENV-1, DENV-2, and DENV-4 by real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction. The effect of...
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| Veröffentlicht in: | Talanta (Oxford) 2018-12, Vol.190, p.391-396 |
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| creator | Kim, Jong-Gil Baek, Seung Hoon Kim, Seungrok Kim, Hae In Lee, Seung Woo Phan, Le Minh Tu Kailasa, Suresh Kumar Park, Tae Jung |
| description | Dengue virus (DENV) is one of the life-threatening viruses to the human. In this study, we have designed specific novel primers for rapid discriminative detection of DENV-1, DENV-2, and DENV-4 by real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction. The effect of parameters such as reaction temperature and magnesium sulfate was investigated on the RT-LAMP reaction for detection of DENV RNA. Under the optimal conditions, this method is able to differentiate and to detect DENV within 25 min, exhibiting detection limit of 3.5 copies/μL. Importantly, the novel specific primers-based RT-LAMP assay did not react with other viruses, suggesting the selectivity of the method towards DENV RNA. The RT-LAMP reaction products are easily visualized with naked-eye when irradiated them under UV light at 365 nm. Amplification products could be visualized directly for color changes. This method provides a facile, and accurate molecular amplication technique for the rapid discriminative detection of dengue viruses. The RT-LAMP platform can be used as a promissing diagnostic tool for discriminative detection of DENV without aid of complicated protocols or sophisticated equipment.
[Display omitted]
•RT-LAMP assay by newly designed primers for Dengue virus detection.•Rapid platform for discriminative detection of Dengue virus.•Detection limit of 3.5 copies/μL within only 25 min.•Four primers used in RT-LAMP assay for each molecular detection. |
| doi_str_mv | 10.1016/j.talanta.2018.08.019 |
| format | Article |
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[Display omitted]
•RT-LAMP assay by newly designed primers for Dengue virus detection.•Rapid platform for discriminative detection of Dengue virus.•Detection limit of 3.5 copies/μL within only 25 min.•Four primers used in RT-LAMP assay for each molecular detection.</description><identifier>ISSN: 0039-9140</identifier><identifier>EISSN: 1873-3573</identifier><identifier>DOI: 10.1016/j.talanta.2018.08.019</identifier><identifier>PMID: 30172524</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Aedes aegypi mosquito ; Dengue virus ; Dengue Virus - genetics ; Dengue Virus - isolation & purification ; Limit of Detection ; Molecular diagnosis ; Nucleic Acid Amplification Techniques - methods ; Reverse Transcription ; Reverse trascription loop-mediated isothermal amplification ; RNA, Viral - genetics ; Specific primers ; Time Factors</subject><ispartof>Talanta (Oxford), 2018-12, Vol.190, p.391-396</ispartof><rights>2018 Elsevier B.V.</rights><rights>Copyright © 2018 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c365t-e9974db1c61fbbae0ec99c87182a84998c0f8305e9c1175b43c16708a34b0a9c3</citedby><cites>FETCH-LOGICAL-c365t-e9974db1c61fbbae0ec99c87182a84998c0f8305e9c1175b43c16708a34b0a9c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.talanta.2018.08.019$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30172524$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kim, Jong-Gil</creatorcontrib><creatorcontrib>Baek, Seung Hoon</creatorcontrib><creatorcontrib>Kim, Seungrok</creatorcontrib><creatorcontrib>Kim, Hae In</creatorcontrib><creatorcontrib>Lee, Seung Woo</creatorcontrib><creatorcontrib>Phan, Le Minh Tu</creatorcontrib><creatorcontrib>Kailasa, Suresh Kumar</creatorcontrib><creatorcontrib>Park, Tae Jung</creatorcontrib><title>Rapid discriminative detection of dengue viruses via loop mediated isothermal amplification</title><title>Talanta (Oxford)</title><addtitle>Talanta</addtitle><description>Dengue virus (DENV) is one of the life-threatening viruses to the human. In this study, we have designed specific novel primers for rapid discriminative detection of DENV-1, DENV-2, and DENV-4 by real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction. The effect of parameters such as reaction temperature and magnesium sulfate was investigated on the RT-LAMP reaction for detection of DENV RNA. Under the optimal conditions, this method is able to differentiate and to detect DENV within 25 min, exhibiting detection limit of 3.5 copies/μL. Importantly, the novel specific primers-based RT-LAMP assay did not react with other viruses, suggesting the selectivity of the method towards DENV RNA. The RT-LAMP reaction products are easily visualized with naked-eye when irradiated them under UV light at 365 nm. Amplification products could be visualized directly for color changes. This method provides a facile, and accurate molecular amplication technique for the rapid discriminative detection of dengue viruses. The RT-LAMP platform can be used as a promissing diagnostic tool for discriminative detection of DENV without aid of complicated protocols or sophisticated equipment.
[Display omitted]
•RT-LAMP assay by newly designed primers for Dengue virus detection.•Rapid platform for discriminative detection of Dengue virus.•Detection limit of 3.5 copies/μL within only 25 min.•Four primers used in RT-LAMP assay for each molecular detection.</description><subject>Aedes aegypi mosquito</subject><subject>Dengue virus</subject><subject>Dengue Virus - genetics</subject><subject>Dengue Virus - isolation & purification</subject><subject>Limit of Detection</subject><subject>Molecular diagnosis</subject><subject>Nucleic Acid Amplification Techniques - methods</subject><subject>Reverse Transcription</subject><subject>Reverse trascription loop-mediated isothermal amplification</subject><subject>RNA, Viral - genetics</subject><subject>Specific primers</subject><subject>Time Factors</subject><issn>0039-9140</issn><issn>1873-3573</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkM1LxDAQxYMo7rr6Jyg9euk6afqRnETEL1gQRE8eQppONUvb1CRd8L83y65ehYHHwHszvB8h5xSWFGh5tV4G1akhqGUGlC8hDhUHZE55xVJWVOyQzAGYSAXNYUZOvF8DQMaAHZMZA1plRZbPyfuLGk2TNMZrZ3ozqGA2mDQYUAdjh8S2cRk-Jkw2xk0efVSVdNaOSY-NUQGbxHgbPtH1qktUP3amNVptw6fkqFWdx7O9Lsjb_d3r7WO6en54ur1ZpZqVRUhRiCpvaqpL2ta1QkAthOYV5ZniuRBcQ8sZFCg0pVVR50zTsgKuWF6DEpotyOXu7ujs14Q-yD7WwS7iQTt5mYEQkQTwIlqLnVU7673DVo6xtnLfkoLccpVruecqt1wlxKEi5i72L6Y69v5L_YKMhuudAWPRjUEnvTY46MjIRZSyseafFz8W041P</recordid><startdate>20181201</startdate><enddate>20181201</enddate><creator>Kim, Jong-Gil</creator><creator>Baek, Seung Hoon</creator><creator>Kim, Seungrok</creator><creator>Kim, Hae In</creator><creator>Lee, Seung Woo</creator><creator>Phan, Le Minh Tu</creator><creator>Kailasa, Suresh Kumar</creator><creator>Park, Tae Jung</creator><general>Elsevier B.V</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>7X8</scope></search><sort><creationdate>20181201</creationdate><title>Rapid discriminative detection of dengue viruses via loop mediated isothermal amplification</title><author>Kim, Jong-Gil ; Baek, Seung Hoon ; Kim, Seungrok ; Kim, Hae In ; Lee, Seung Woo ; Phan, Le Minh Tu ; Kailasa, Suresh Kumar ; Park, Tae Jung</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c365t-e9974db1c61fbbae0ec99c87182a84998c0f8305e9c1175b43c16708a34b0a9c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Aedes aegypi mosquito</topic><topic>Dengue virus</topic><topic>Dengue Virus - genetics</topic><topic>Dengue Virus - isolation & purification</topic><topic>Limit of Detection</topic><topic>Molecular diagnosis</topic><topic>Nucleic Acid Amplification Techniques - methods</topic><topic>Reverse Transcription</topic><topic>Reverse trascription loop-mediated isothermal amplification</topic><topic>RNA, Viral - genetics</topic><topic>Specific primers</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kim, Jong-Gil</creatorcontrib><creatorcontrib>Baek, Seung Hoon</creatorcontrib><creatorcontrib>Kim, Seungrok</creatorcontrib><creatorcontrib>Kim, Hae In</creatorcontrib><creatorcontrib>Lee, Seung Woo</creatorcontrib><creatorcontrib>Phan, Le Minh Tu</creatorcontrib><creatorcontrib>Kailasa, Suresh Kumar</creatorcontrib><creatorcontrib>Park, Tae Jung</creatorcontrib><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><jtitle>Talanta (Oxford)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kim, Jong-Gil</au><au>Baek, Seung Hoon</au><au>Kim, Seungrok</au><au>Kim, Hae In</au><au>Lee, Seung Woo</au><au>Phan, Le Minh Tu</au><au>Kailasa, Suresh Kumar</au><au>Park, Tae Jung</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Rapid discriminative detection of dengue viruses via loop mediated isothermal amplification</atitle><jtitle>Talanta (Oxford)</jtitle><addtitle>Talanta</addtitle><date>2018-12-01</date><risdate>2018</risdate><volume>190</volume><spage>391</spage><epage>396</epage><pages>391-396</pages><issn>0039-9140</issn><eissn>1873-3573</eissn><abstract>Dengue virus (DENV) is one of the life-threatening viruses to the human. In this study, we have designed specific novel primers for rapid discriminative detection of DENV-1, DENV-2, and DENV-4 by real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction. The effect of parameters such as reaction temperature and magnesium sulfate was investigated on the RT-LAMP reaction for detection of DENV RNA. Under the optimal conditions, this method is able to differentiate and to detect DENV within 25 min, exhibiting detection limit of 3.5 copies/μL. Importantly, the novel specific primers-based RT-LAMP assay did not react with other viruses, suggesting the selectivity of the method towards DENV RNA. The RT-LAMP reaction products are easily visualized with naked-eye when irradiated them under UV light at 365 nm. Amplification products could be visualized directly for color changes. This method provides a facile, and accurate molecular amplication technique for the rapid discriminative detection of dengue viruses. The RT-LAMP platform can be used as a promissing diagnostic tool for discriminative detection of DENV without aid of complicated protocols or sophisticated equipment.
[Display omitted]
•RT-LAMP assay by newly designed primers for Dengue virus detection.•Rapid platform for discriminative detection of Dengue virus.•Detection limit of 3.5 copies/μL within only 25 min.•Four primers used in RT-LAMP assay for each molecular detection.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>30172524</pmid><doi>10.1016/j.talanta.2018.08.019</doi><tpages>6</tpages></addata></record> |
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| subjects | Aedes aegypi mosquito Dengue virus Dengue Virus - genetics Dengue Virus - isolation & purification Limit of Detection Molecular diagnosis Nucleic Acid Amplification Techniques - methods Reverse Transcription Reverse trascription loop-mediated isothermal amplification RNA, Viral - genetics Specific primers Time Factors |
| title | Rapid discriminative detection of dengue viruses via loop mediated isothermal amplification |
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