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Detection of Zika virus using reverse-transcription LAMP coupled with reverse dot blot analysis in saliva

Identifieur interne : 000603 ( Pmc/Checkpoint ); précédent : 000602; suivant : 000604

Detection 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 :

RBID : PMC:5798782

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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PMC:5798782

Le document en format XML

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<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>
<author>
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</author>
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</author>
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</author>
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<article-id pub-id-type="pmc">5798782</article-id>
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<subj-group>
<subject>Molecular Probe Techniques</subject>
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<subject>Biochemistry</subject>
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<subject>Nucleic Acids</subject>
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<article-title>Detection of Zika virus using reverse-transcription LAMP coupled with reverse dot blot analysis in saliva</article-title>
<alt-title alt-title-type="running-head">Diagnostics for emerging infectious diseases: Zika virus</alt-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<contrib-id authenticated="true" contrib-id-type="orcid">http://orcid.org/0000-0002-5669-8749</contrib-id>
<name>
<surname>Sabalza</surname>
<given-names>Maite</given-names>
</name>
<role content-type="http://credit.casrai.org/">Conceptualization</role>
<role content-type="http://credit.casrai.org/">Investigation</role>
<role content-type="http://credit.casrai.org/">Visualization</role>
<role content-type="http://credit.casrai.org/">Writing – original draft</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
<xref ref-type="corresp" rid="cor001">*</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Yasmin</surname>
<given-names>Rubina</given-names>
</name>
<role content-type="http://credit.casrai.org/">Conceptualization</role>
<role content-type="http://credit.casrai.org/">Investigation</role>
<role content-type="http://credit.casrai.org/">Visualization</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff002">
<sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Barber</surname>
<given-names>Cheryl A.</given-names>
</name>
<role content-type="http://credit.casrai.org/">Investigation</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Castro</surname>
<given-names>Talita</given-names>
</name>
<role content-type="http://credit.casrai.org/">Investigation</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
<xref ref-type="aff" rid="aff003">
<sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Malamud</surname>
<given-names>Daniel</given-names>
</name>
<role content-type="http://credit.casrai.org/">Conceptualization</role>
<role content-type="http://credit.casrai.org/">Supervision</role>
<role content-type="http://credit.casrai.org/">Visualization</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
<xref ref-type="aff" rid="aff004">
<sup>4</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Kim</surname>
<given-names>Beum Jun</given-names>
</name>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff002">
<sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Zhu</surname>
<given-names>Hui</given-names>
</name>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff002">
<sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Montagna</surname>
<given-names>Richard A.</given-names>
</name>
<role content-type="http://credit.casrai.org/">Conceptualization</role>
<role content-type="http://credit.casrai.org/">Funding acquisition</role>
<role content-type="http://credit.casrai.org/">Project administration</role>
<role content-type="http://credit.casrai.org/">Resources</role>
<role content-type="http://credit.casrai.org/">Supervision</role>
<role content-type="http://credit.casrai.org/">Visualization</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff002">
<sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Abrams</surname>
<given-names>William R.</given-names>
</name>
<role content-type="http://credit.casrai.org/">Conceptualization</role>
<role content-type="http://credit.casrai.org/">Formal analysis</role>
<role content-type="http://credit.casrai.org/">Visualization</role>
<role content-type="http://credit.casrai.org/">Writing – original draft</role>
<role content-type="http://credit.casrai.org/">Writing – review & editing</role>
<xref ref-type="aff" rid="aff001">
<sup>1</sup>
</xref>
</contrib>
</contrib-group>
<aff id="aff001">
<label>1</label>
<addr-line>Department of Basic Sciences, New York University College of Dentistry, New York, New York, United States of America</addr-line>
</aff>
<aff id="aff002">
<label>2</label>
<addr-line>Rheonix, Inc., Ithaca, New York, United States of America</addr-line>
</aff>
<aff id="aff003">
<label>3</label>
<addr-line>Stomatology Department, School of Dentistry, University of São Paulo, São Paulo, São Paulo, Brazil</addr-line>
</aff>
<aff id="aff004">
<label>4</label>
<addr-line>Department of Medicine, New York University School of Medicine, New York, New York, United States of America</addr-line>
</aff>
<contrib-group>
<contrib contrib-type="editor">
<name>
<surname>Kalendar</surname>
<given-names>Ruslan</given-names>
</name>
<role>Editor</role>
<xref ref-type="aff" rid="edit1"></xref>
</contrib>
</contrib-group>
<aff id="edit1">
<addr-line>University of Helsinki, FINLAND</addr-line>
</aff>
<author-notes>
<fn fn-type="COI-statement" id="coi001">
<p>
<bold>Competing Interests: </bold>
We have the following interests: R.M., R.Y., B.J.K. and H.Z. are employed by Rheonix, Inc. In addition, each own shares and/or options to purchase shares in Rheonix, Inc.. The Encompass Optimum workstation and Rheonix CARD
<sup>®</sup>
cartridge are produced by Rheonix, Inc. There are no patents, further products in development or further marketed products to declare. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials.</p>
</fn>
<corresp id="cor001">* E-mail:
<email>ms10129@nyu.edu</email>
</corresp>
</author-notes>
<pub-date pub-type="epub">
<day>5</day>
<month>2</month>
<year>2018</year>
</pub-date>
<pub-date pub-type="collection">
<year>2018</year>
</pub-date>
<volume>13</volume>
<issue>2</issue>
<elocation-id>e0192398</elocation-id>
<history>
<date date-type="received">
<day>23</day>
<month>8</month>
<year>2017</year>
</date>
<date date-type="accepted">
<day>23</day>
<month>1</month>
<year>2018</year>
</date>
</history>
<permissions>
<copyright-statement>© 2018 Sabalza et al</copyright-statement>
<copyright-year>2018</copyright-year>
<copyright-holder>Sabalza et al</copyright-holder>
<license xlink:href="http://creativecommons.org/licenses/by/4.0/">
<license-p>This is an open access article distributed under the terms of the
<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License</ext-link>
, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</license-p>
</license>
</permissions>
<self-uri content-type="pdf" xlink:href="pone.0192398.pdf"></self-uri>
<abstract>
<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>
</abstract>
<funding-group>
<award-group id="award001">
<funding-source>
<institution-wrap>
<institution-id institution-id-type="funder-id">http://dx.doi.org/10.13039/100000072</institution-id>
<institution>National Institute of Dental and Craniofacial Research</institution>
</institution-wrap>
</funding-source>
<award-id>R44DE024456 and 3R44DE024456-03S1</award-id>
</award-group>
<funding-statement>Research reported in this publication was supported by the National Institute of Dental and Craniofacial Research of the National Institutes of Health under Award Numbers R44DE024456 and 3R44DE024456-03S1. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Four of the authors (Richard Montagna, Rubina Yasmin, Beum Jun Kim and Hui Zhu) are employees of Rheonix, Inc. Rheonix, Inc. provided support in the form of salaries for authors RM, RY, BJK and HZ, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the "author contributions" section.</funding-statement>
</funding-group>
<counts>
<fig-count count="5"></fig-count>
<table-count count="5"></table-count>
<page-count count="17"></page-count>
</counts>
<custom-meta-group>
<custom-meta id="data-availability">
<meta-name>Data Availability</meta-name>
<meta-value>All relevant data are within the paper.</meta-value>
</custom-meta>
</custom-meta-group>
</article-meta>
<notes>
<title>Data Availability</title>
<p>All relevant data are within the paper.</p>
</notes>
</front>
</pmc>
<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>
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<name sortKey="Abrams, William R" sort="Abrams, William R" uniqKey="Abrams W" first="William R." last="Abrams">William R. Abrams</name>
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<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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