Colorimetric Detection of Dengue by Single Tube Reverse-Transcription-Loop-Mediated Isothermal Amplification
Dengue is usually diagnosed by isolation of the virus, serology or molecular diagnostic methods. Several commercial kits for the diagnosis of dengue are existing, but concerns have arisen regarding to the affordability and performance characteristics of these kits. Hence, the loop-mediated isotherma...
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| Veröffentlicht in: | PloS one 2015-09, Vol.10 (9), p.e0138694-e0138694 |
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| creator | Lau, Yee-Ling Lai, Meng-Yee Teoh, Boon-Teong Abd-Jamil, Juraina Johari, Jefree Sam, Sing-Sin Tan, Kim-Kee AbuBakar, Sazaly |
| description | Dengue is usually diagnosed by isolation of the virus, serology or molecular diagnostic methods. Several commercial kits for the diagnosis of dengue are existing, but concerns have arisen regarding to the affordability and performance characteristics of these kits. Hence, the loop-mediated isothermal amplification (LAMP) is potentially ideal to be used especially in resource limited environments. Serum was collected from healthy donors and patients diagnosed with dengue infection. RNA extracted from the serum samples were tested by reverse-transcription-LAMP assay developed based on 3'-NCR gene sequences for DENV 1-4. Results were interpreted by a turbidity meter in real time or visually at the end of the assay. Sensitivity and specificity of RT-LAMP results were calculated and compared to qRT-PCR and ELISA. RT-LAMP is highly sensitive with the detection limit of 10 RNA copies for all serotypes. Dengue virus RNA was detected in all positive samples using RT-LAMP and none of the negative samples within 30-45 minutes. With continuing efforts in the optimization of this assay, RT-LAMP may provide a simple and reliable test for detecting DENV in areas where dengue is prevalent. |
| doi_str_mv | 10.1371/journal.pone.0138694 |
| format | Article |
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Several commercial kits for the diagnosis of dengue are existing, but concerns have arisen regarding to the affordability and performance characteristics of these kits. Hence, the loop-mediated isothermal amplification (LAMP) is potentially ideal to be used especially in resource limited environments. Serum was collected from healthy donors and patients diagnosed with dengue infection. RNA extracted from the serum samples were tested by reverse-transcription-LAMP assay developed based on 3'-NCR gene sequences for DENV 1-4. Results were interpreted by a turbidity meter in real time or visually at the end of the assay. Sensitivity and specificity of RT-LAMP results were calculated and compared to qRT-PCR and ELISA. RT-LAMP is highly sensitive with the detection limit of 10 RNA copies for all serotypes. Dengue virus RNA was detected in all positive samples using RT-LAMP and none of the negative samples within 30-45 minutes. With continuing efforts in the optimization of this assay, RT-LAMP may provide a simple and reliable test for detecting DENV in areas where dengue is prevalent.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0138694</identifier><identifier>PMID: 26384248</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Assaying ; Case-Control Studies ; Chikungunya virus ; Colorimetry ; Colorimetry - methods ; Dengue ; Dengue - blood ; Dengue - diagnosis ; Dengue fever ; Deoxyribonucleic acid ; Development and progression ; Diagnosis ; Diagnostic systems ; Diagnostic tests ; DNA ; Education ; Encephalitis ; Enzyme-linked immunosorbent assay ; Ethics ; Gene sequencing ; Genetic aspects ; Humans ; Immunoglobulins ; Infections ; Infectious diseases ; Medicine ; Methods ; Nucleic Acid Amplification Techniques - methods ; Optimization ; Parasitology ; Patient outcomes ; Patients ; Ribonucleic acid ; RNA ; RNA viruses ; Sensitivity and Specificity ; Serology ; Serotypes ; Surveillance ; Transcription (Genetics) ; Turbidity ; Vector-borne diseases ; Viral diseases ; Viruses ; West Nile virus</subject><ispartof>PloS one, 2015-09, Vol.10 (9), p.e0138694-e0138694</ispartof><rights>COPYRIGHT 2015 Public Library of Science</rights><rights>2015 Lau et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2015 Lau et al 2015 Lau et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-254caad4fd38682dffee9bb4625f6cec33b46bb19359d178201bbe26253c754b3</citedby><cites>FETCH-LOGICAL-c692t-254caad4fd38682dffee9bb4625f6cec33b46bb19359d178201bbe26253c754b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4575147/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4575147/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79342,79343</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26384248$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lau, Yee-Ling</creatorcontrib><creatorcontrib>Lai, Meng-Yee</creatorcontrib><creatorcontrib>Teoh, Boon-Teong</creatorcontrib><creatorcontrib>Abd-Jamil, Juraina</creatorcontrib><creatorcontrib>Johari, Jefree</creatorcontrib><creatorcontrib>Sam, Sing-Sin</creatorcontrib><creatorcontrib>Tan, Kim-Kee</creatorcontrib><creatorcontrib>AbuBakar, Sazaly</creatorcontrib><title>Colorimetric Detection of Dengue by Single Tube Reverse-Transcription-Loop-Mediated Isothermal Amplification</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Dengue is usually diagnosed by isolation of the virus, serology or molecular diagnostic methods. Several commercial kits for the diagnosis of dengue are existing, but concerns have arisen regarding to the affordability and performance characteristics of these kits. Hence, the loop-mediated isothermal amplification (LAMP) is potentially ideal to be used especially in resource limited environments. Serum was collected from healthy donors and patients diagnosed with dengue infection. RNA extracted from the serum samples were tested by reverse-transcription-LAMP assay developed based on 3'-NCR gene sequences for DENV 1-4. Results were interpreted by a turbidity meter in real time or visually at the end of the assay. Sensitivity and specificity of RT-LAMP results were calculated and compared to qRT-PCR and ELISA. RT-LAMP is highly sensitive with the detection limit of 10 RNA copies for all serotypes. Dengue virus RNA was detected in all positive samples using RT-LAMP and none of the negative samples within 30-45 minutes. With continuing efforts in the optimization of this assay, RT-LAMP may provide a simple and reliable test for detecting DENV in areas where dengue is prevalent.</description><subject>Assaying</subject><subject>Case-Control Studies</subject><subject>Chikungunya virus</subject><subject>Colorimetry</subject><subject>Colorimetry - methods</subject><subject>Dengue</subject><subject>Dengue - blood</subject><subject>Dengue - diagnosis</subject><subject>Dengue fever</subject><subject>Deoxyribonucleic acid</subject><subject>Development and progression</subject><subject>Diagnosis</subject><subject>Diagnostic systems</subject><subject>Diagnostic tests</subject><subject>DNA</subject><subject>Education</subject><subject>Encephalitis</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Ethics</subject><subject>Gene sequencing</subject><subject>Genetic aspects</subject><subject>Humans</subject><subject>Immunoglobulins</subject><subject>Infections</subject><subject>Infectious diseases</subject><subject>Medicine</subject><subject>Methods</subject><subject>Nucleic Acid Amplification Techniques - methods</subject><subject>Optimization</subject><subject>Parasitology</subject><subject>Patient outcomes</subject><subject>Patients</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA viruses</subject><subject>Sensitivity and Specificity</subject><subject>Serology</subject><subject>Serotypes</subject><subject>Surveillance</subject><subject>Transcription (Genetics)</subject><subject>Turbidity</subject><subject>Vector-borne diseases</subject><subject>Viral diseases</subject><subject>Viruses</subject><subject>West Nile virus</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNk11v0zAUhiMEYqPwDxBEQkJwkRJ_5OsGqSpflYombYVby3FOUldOnNnOxP49zppNDdoF8kUc-znv8XntEwSvUbxEJEOfDnowHVfLXnewjBHJ04I-Cc5RQXCU4pg8PZmfBS-sPcRx4qn0eXCGU5JTTPPzQK210ka24IwU4RdwIJzUXahr_9M1A4TlbXglu0ZBuBtKCC_hBoyFaGd4Z4WR_YhHW6376CdUkjuowo3Vbg-m5Spctb2StRR8xF4Gz2quLLyavovg17evu_WPaHvxfbNebSORFthFOKGC84rWlT9ujqu6BijKkqY4qVMBghA_L0tfXVJUKMtxjMoSsN8mIktoSRbB26Nur7Rlk1GWoQwVOM9HQxbB5khUmh9Y7w3g5pZpLtndgjYN48ZJoYClFEBQEVMogSaC51laJTXGHKVFznPstT5P2YayhUpA5wxXM9H5Tif3rNE3jCZZgmjmBT5MAkZfD2Ada6UVoBTvQA93504LUhA65nr3D_p4dRPVcF-A7Grt84pRlK0oLuKkyCnx1PIRyo8KWin8q6qlX58FfJwFeMbBH9fwwVq2ubr8f_bi95x9f8LugSu3t1oN45Oxc5AeQWG0tQbqB5NRzMamuHeDjU3BpqbwYW9OL-gh6L4LyF9Gbghr</recordid><startdate>20150918</startdate><enddate>20150918</enddate><creator>Lau, Yee-Ling</creator><creator>Lai, Meng-Yee</creator><creator>Teoh, Boon-Teong</creator><creator>Abd-Jamil, Juraina</creator><creator>Johari, Jefree</creator><creator>Sam, Sing-Sin</creator><creator>Tan, Kim-Kee</creator><creator>AbuBakar, Sazaly</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>COVID</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20150918</creationdate><title>Colorimetric Detection of Dengue by Single Tube Reverse-Transcription-Loop-Mediated Isothermal Amplification</title><author>Lau, Yee-Ling ; Lai, Meng-Yee ; Teoh, Boon-Teong ; Abd-Jamil, Juraina ; Johari, Jefree ; Sam, Sing-Sin ; Tan, Kim-Kee ; AbuBakar, Sazaly</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-254caad4fd38682dffee9bb4625f6cec33b46bb19359d178201bbe26253c754b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Assaying</topic><topic>Case-Control Studies</topic><topic>Chikungunya virus</topic><topic>Colorimetry</topic><topic>Colorimetry - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lau, Yee-Ling</au><au>Lai, Meng-Yee</au><au>Teoh, Boon-Teong</au><au>Abd-Jamil, Juraina</au><au>Johari, Jefree</au><au>Sam, Sing-Sin</au><au>Tan, Kim-Kee</au><au>AbuBakar, Sazaly</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Colorimetric Detection of Dengue by Single Tube Reverse-Transcription-Loop-Mediated Isothermal Amplification</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2015-09-18</date><risdate>2015</risdate><volume>10</volume><issue>9</issue><spage>e0138694</spage><epage>e0138694</epage><pages>e0138694-e0138694</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Dengue is usually diagnosed by isolation of the virus, serology or molecular diagnostic methods. Several commercial kits for the diagnosis of dengue are existing, but concerns have arisen regarding to the affordability and performance characteristics of these kits. Hence, the loop-mediated isothermal amplification (LAMP) is potentially ideal to be used especially in resource limited environments. Serum was collected from healthy donors and patients diagnosed with dengue infection. RNA extracted from the serum samples were tested by reverse-transcription-LAMP assay developed based on 3'-NCR gene sequences for DENV 1-4. Results were interpreted by a turbidity meter in real time or visually at the end of the assay. Sensitivity and specificity of RT-LAMP results were calculated and compared to qRT-PCR and ELISA. RT-LAMP is highly sensitive with the detection limit of 10 RNA copies for all serotypes. Dengue virus RNA was detected in all positive samples using RT-LAMP and none of the negative samples within 30-45 minutes. With continuing efforts in the optimization of this assay, RT-LAMP may provide a simple and reliable test for detecting DENV in areas where dengue is prevalent.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>26384248</pmid><doi>10.1371/journal.pone.0138694</doi><oa>free_for_read</oa></addata></record> |
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| subjects | Assaying Case-Control Studies Chikungunya virus Colorimetry Colorimetry - methods Dengue Dengue - blood Dengue - diagnosis Dengue fever Deoxyribonucleic acid Development and progression Diagnosis Diagnostic systems Diagnostic tests DNA Education Encephalitis Enzyme-linked immunosorbent assay Ethics Gene sequencing Genetic aspects Humans Immunoglobulins Infections Infectious diseases Medicine Methods Nucleic Acid Amplification Techniques - methods Optimization Parasitology Patient outcomes Patients Ribonucleic acid RNA RNA viruses Sensitivity and Specificity Serology Serotypes Surveillance Transcription (Genetics) Turbidity Vector-borne diseases Viral diseases Viruses West Nile virus |
| title | Colorimetric Detection of Dengue by Single Tube Reverse-Transcription-Loop-Mediated Isothermal Amplification |
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