Detection of Zika virus using reverse-transcription LAMP coupled with reverse dot blot analysis in saliva
Identifieur interne : 000A11 ( Main/Exploration ); précédent : 000A10; suivant : 000A12Detection of Zika virus using reverse-transcription LAMP coupled with reverse dot blot analysis in saliva
Auteurs : Maite Sabalza [États-Unis] ; Rubina Yasmin [États-Unis] ; Cheryl A. Barber [États-Unis] ; Talita Castro [États-Unis, Brésil] ; Daniel Malamud [États-Unis] ; Beum Jun Kim [États-Unis] ; Hui Zhu [États-Unis] ; Richard A. Montagna [États-Unis] ; William R. Abrams [États-Unis]Source :
- PLoS ONE [ 1932-6203 ] ; 2018.
Descripteurs français
- KwdFr :
- MESH :
- génétique : ARN viral, Virus Zika.
- isolement et purification : Virus Zika.
- virologie : Salive.
- Humains, Techniques d'amplification d'acides nucléiques, Transcription inverse.
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : RNA, Viral.
- genetics : Zika Virus.
- isolation & purification : Zika Virus.
- methods : Nucleic Acid Amplification Techniques.
- virology : Saliva.
- Humans, Reverse Transcription.
Abstract
In recent years, there have been increasing numbers of infectious disease outbreaks that spread rapidly to population centers resulting from global travel, population vulnerabilities, environmental factors, and ecological disasters such as floods and earthquakes. Some examples of the recent outbreaks are the Ebola epidemic in West Africa, Middle East respiratory syndrome coronavirus (MERS-Co) in the Middle East, and the Zika outbreak through the Americas. We have created a generic protocol for detection of pathogen RNA and/or DNA using loop-mediated isothermal amplification (LAMP) and reverse dot-blot for detection (RDB) and processed automatically in a microfluidic device. In particular, we describe how a microfluidic assay to detect HIV viral RNA was converted to detect Zika virus (ZIKV) RNA. We first optimized the RT-LAMP assay to detect ZIKV RNA using a benchtop isothermal amplification device. Then we implemented the assay in a microfluidic device that will allow analyzing 24 samples simultaneously and automatically from sample introduction to detection by RDB technique. Preliminary data using saliva samples spiked with ZIKV showed that our diagnostic system detects ZIKV RNA in saliva. These results will be validated in further experiments with well-characterized ZIKV human specimens of saliva. The described strategy and methodology to convert the HIV diagnostic assay and platform to a ZIKV RNA detection assay provides a model that can be readily utilized for detection of the next emerging or re-emerging infectious disease.
Url:
DOI: 10.1371/journal.pone.0192398
PubMed: 29401479
PubMed Central: 5798782
Affiliations:
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Le document en format XML
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<front><div type="abstract" xml:lang="en"><p>In recent years, there have been increasing numbers of infectious disease outbreaks that spread rapidly to population centers resulting from global travel, population vulnerabilities, environmental factors, and ecological disasters such as floods and earthquakes. Some examples of the recent outbreaks are the Ebola epidemic in West Africa, Middle East respiratory syndrome coronavirus (MERS-Co) in the Middle East, and the Zika outbreak through the Americas. We have created a generic protocol for detection of pathogen RNA and/or DNA using loop-mediated isothermal amplification (LAMP) and reverse dot-blot for detection (RDB) and processed automatically in a microfluidic device. In particular, we describe how a microfluidic assay to detect HIV viral RNA was converted to detect Zika virus (ZIKV) RNA. We first optimized the RT-LAMP assay to detect ZIKV RNA using a benchtop isothermal amplification device. Then we implemented the assay in a microfluidic device that will allow analyzing 24 samples simultaneously and automatically from sample introduction to detection by RDB technique. Preliminary data using saliva samples spiked with ZIKV showed that our diagnostic system detects ZIKV RNA in saliva. These results will be validated in further experiments with well-characterized ZIKV human specimens of saliva. The described strategy and methodology to convert the HIV diagnostic assay and platform to a ZIKV RNA detection assay provides a model that can be readily utilized for detection of the next emerging or re-emerging infectious disease.</p>
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<author><name sortKey="Bretzel, G" uniqKey="Bretzel G">G Bretzel</name>
</author>
<author><name sortKey="Hoelscher, M" uniqKey="Hoelscher M">M Hoelscher</name>
</author>
</analytic>
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</author>
<author><name sortKey="Supasa, P" uniqKey="Supasa P">P Supasa</name>
</author>
<author><name sortKey="Wongwiwat, W" uniqKey="Wongwiwat W">W Wongwiwat</name>
</author>
<author><name sortKey="Rouvinski, A" uniqKey="Rouvinski A">A Rouvinski</name>
</author>
<author><name sortKey="Barba Spaeth, G" uniqKey="Barba Spaeth G">G Barba-Spaeth</name>
</author>
<author><name sortKey="Duangchinda, T" uniqKey="Duangchinda T">T Duangchinda</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Sabalza, M" uniqKey="Sabalza M">M Sabalza</name>
</author>
<author><name sortKey="Barber, Ca" uniqKey="Barber C">CA Barber</name>
</author>
<author><name sortKey="Abrams, Wr" uniqKey="Abrams W">WR Abrams</name>
</author>
<author><name sortKey="Montagna, R" uniqKey="Montagna R">R Montagna</name>
</author>
<author><name sortKey="Malamud, D" uniqKey="Malamud D">D Malamud</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Chotiwan, N" uniqKey="Chotiwan N">N Chotiwan</name>
</author>
<author><name sortKey="Brewster, Cd" uniqKey="Brewster C">CD Brewster</name>
</author>
<author><name sortKey="Magalhaes, T" uniqKey="Magalhaes T">T Magalhaes</name>
</author>
<author><name sortKey="Weger Lucarelli, J" uniqKey="Weger Lucarelli J">J Weger-Lucarelli</name>
</author>
<author><name sortKey="Duggal, Nk" uniqKey="Duggal N">NK Duggal</name>
</author>
<author><name sortKey="Ruckert, C" uniqKey="Ruckert C">C Rückert</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Song, J" uniqKey="Song J">J Song</name>
</author>
<author><name sortKey="Mauk, Mg" uniqKey="Mauk M">MG Mauk</name>
</author>
<author><name sortKey="Hackett, Ba" uniqKey="Hackett B">BA Hackett</name>
</author>
<author><name sortKey="Cherry, S" uniqKey="Cherry S">S Cherry</name>
</author>
<author><name sortKey="Bau, Hh" uniqKey="Bau H">HH Bau</name>
</author>
<author><name sortKey="Liu, C" uniqKey="Liu C">C Liu</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Nicolini, Am" uniqKey="Nicolini A">AM Nicolini</name>
</author>
<author><name sortKey="Mccracken, Ke" uniqKey="Mccracken K">KE McCracken</name>
</author>
<author><name sortKey="Yoon, J Y" uniqKey="Yoon J">J-Y Yoon</name>
</author>
</analytic>
</biblStruct>
<biblStruct></biblStruct>
</listBibl>
</div1>
</back>
</TEI>
<affiliations><list><country><li>Brésil</li>
<li>États-Unis</li>
</country>
<region><li>État de New York</li>
<li>État de São Paulo</li>
</region>
<settlement><li>São Paulo</li>
</settlement>
<orgName><li>Université de São Paulo</li>
</orgName>
</list>
<tree><country name="États-Unis"><region name="État de New York"><name sortKey="Sabalza, Maite" sort="Sabalza, Maite" uniqKey="Sabalza M" first="Maite" last="Sabalza">Maite Sabalza</name>
</region>
<name sortKey="Abrams, William R" sort="Abrams, William R" uniqKey="Abrams W" first="William R." last="Abrams">William R. Abrams</name>
<name sortKey="Barber, Cheryl A" sort="Barber, Cheryl A" uniqKey="Barber C" first="Cheryl A." last="Barber">Cheryl A. Barber</name>
<name sortKey="Castro, Talita" sort="Castro, Talita" uniqKey="Castro T" first="Talita" last="Castro">Talita Castro</name>
<name sortKey="Kim, Beum Jun" sort="Kim, Beum Jun" uniqKey="Kim B" first="Beum Jun" last="Kim">Beum Jun Kim</name>
<name sortKey="Malamud, Daniel" sort="Malamud, Daniel" uniqKey="Malamud D" first="Daniel" last="Malamud">Daniel Malamud</name>
<name sortKey="Malamud, Daniel" sort="Malamud, Daniel" uniqKey="Malamud D" first="Daniel" last="Malamud">Daniel Malamud</name>
<name sortKey="Montagna, Richard A" sort="Montagna, Richard A" uniqKey="Montagna R" first="Richard A." last="Montagna">Richard A. Montagna</name>
<name sortKey="Yasmin, Rubina" sort="Yasmin, Rubina" uniqKey="Yasmin R" first="Rubina" last="Yasmin">Rubina Yasmin</name>
<name sortKey="Zhu, Hui" sort="Zhu, Hui" uniqKey="Zhu H" first="Hui" last="Zhu">Hui Zhu</name>
</country>
<country name="Brésil"><region name="État de São Paulo"><name sortKey="Castro, Talita" sort="Castro, Talita" uniqKey="Castro T" first="Talita" last="Castro">Talita Castro</name>
</region>
</country>
</tree>
</affiliations>
</record>
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