Real-time PCR detection of the HhaI tandem DNA repeat in pre- and post-patent Brugia malayi infections: a study in Indonesian transmigrants
Identifieur interne : 006501 ( Ncbi/Merge ); précédent : 006500; suivant : 006502Real-time PCR detection of the HhaI tandem DNA repeat in pre- and post-patent Brugia malayi infections: a study in Indonesian transmigrants
Auteurs : Anna Albers [Allemagne] ; Erliyani Sartono [Pays-Bas] ; Sitti Wahyuni [Indonésie] ; Maria Yazdanbakhsh [Pays-Bas] ; Rick M. Maizels [Royaume-Uni] ; Ute Klarmann-Schulz [Allemagne] ; Kenneth Pfarr [Allemagne] ; Achim Hoerauf [Allemagne]Source :
- Parasites & Vectors [ 1756-3305 ] ; 2014.
Abstract
Lymphatic filariasis caused by
An optimized real-time PCR targeting the brugian
The PCR detected
The results demonstrate that the
Url:
DOI: 10.1186/1756-3305-7-146
PubMed: 24685183
PubMed Central: 4021971
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PMC:4021971Le document en format XML
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tandem DNA repeat in pre- and post-patent <italic>Brugia malayi</italic>
infections: a study in Indonesian transmigrants</title>
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Real-time PCR detection of the <italic>HhaI</italic>
tandem DNA repeat in pre- and post-patent <italic>Brugia malayi</italic>
infections: a study in Indonesian transmigrants</title>
<author><name sortKey="Albers, Anna" sort="Albers, Anna" uniqKey="Albers A" first="Anna" last="Albers">Anna Albers</name>
<affiliation wicri:level="1"><nlm:aff id="I1">Institute of Medical Microbiology, Immunology and Parasitology, University Hospital of Bonn, Sigmund-Freud-Str. 25, D-53105 Bonn, Germany</nlm:aff>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Institute of Medical Microbiology, Immunology and Parasitology, University Hospital of Bonn, Sigmund-Freud-Str. 25, D-53105 Bonn</wicri:regionArea>
<wicri:noRegion>53105 Bonn</wicri:noRegion>
<wicri:noRegion>53105 Bonn</wicri:noRegion>
<wicri:noRegion>D-53105 Bonn</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Sartono, Erliyani" sort="Sartono, Erliyani" uniqKey="Sartono E" first="Erliyani" last="Sartono">Erliyani Sartono</name>
<affiliation wicri:level="1"><nlm:aff id="I2">Department of Parasitology, Leiden University Medical Center, P.O Box 9600, 2300, RC Leiden, The Netherlands</nlm:aff>
<country xml:lang="fr">Pays-Bas</country>
<wicri:regionArea>Department of Parasitology, Leiden University Medical Center, P.O Box 9600, 2300, RC Leiden</wicri:regionArea>
<wicri:noRegion>RC Leiden</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Wahyuni, Sitti" sort="Wahyuni, Sitti" uniqKey="Wahyuni S" first="Sitti" last="Wahyuni">Sitti Wahyuni</name>
<affiliation wicri:level="1"><nlm:aff id="I3">Parasitology Department, Medical Faculty, Hasanuddin University, Jl. Perintis Kemerdekaan 10 Tamalanrea, Makassar 90245, Indonesia</nlm:aff>
<country xml:lang="fr">Indonésie</country>
<wicri:regionArea>Parasitology Department, Medical Faculty, Hasanuddin University, Jl. Perintis Kemerdekaan 10 Tamalanrea, Makassar 90245</wicri:regionArea>
<wicri:noRegion>Makassar 90245</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Yazdanbakhsh, Maria" sort="Yazdanbakhsh, Maria" uniqKey="Yazdanbakhsh M" first="Maria" last="Yazdanbakhsh">Maria Yazdanbakhsh</name>
<affiliation wicri:level="1"><nlm:aff id="I2">Department of Parasitology, Leiden University Medical Center, P.O Box 9600, 2300, RC Leiden, The Netherlands</nlm:aff>
<country xml:lang="fr">Pays-Bas</country>
<wicri:regionArea>Department of Parasitology, Leiden University Medical Center, P.O Box 9600, 2300, RC Leiden</wicri:regionArea>
<wicri:noRegion>RC Leiden</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Maizels, Rick M" sort="Maizels, Rick M" uniqKey="Maizels R" first="Rick M" last="Maizels">Rick M. Maizels</name>
<affiliation wicri:level="4"><nlm:aff id="I4">Institute for Immunology & Infection Research, University of Edinburgh, Ashworth Laboratories, Edinburgh EH9 3JT, UK</nlm:aff>
<country xml:lang="fr">Royaume-Uni</country>
<wicri:regionArea>Institute for Immunology & Infection Research, University of Edinburgh, Ashworth Laboratories, Edinburgh EH9 3JT</wicri:regionArea>
<orgName type="university">Université d'Édimbourg</orgName>
<placeName><settlement type="city">Édimbourg</settlement>
<region type="country">Écosse</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Klarmann Schulz, Ute" sort="Klarmann Schulz, Ute" uniqKey="Klarmann Schulz U" first="Ute" last="Klarmann-Schulz">Ute Klarmann-Schulz</name>
<affiliation wicri:level="1"><nlm:aff id="I1">Institute of Medical Microbiology, Immunology and Parasitology, University Hospital of Bonn, Sigmund-Freud-Str. 25, D-53105 Bonn, Germany</nlm:aff>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Institute of Medical Microbiology, Immunology and Parasitology, University Hospital of Bonn, Sigmund-Freud-Str. 25, D-53105 Bonn</wicri:regionArea>
<wicri:noRegion>53105 Bonn</wicri:noRegion>
<wicri:noRegion>53105 Bonn</wicri:noRegion>
<wicri:noRegion>D-53105 Bonn</wicri:noRegion>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="I5">Institute for Medical Biometry, Informatics, and Epidemiology, University Hospital of Bonn, Sigmund-Freud-Str. 25, D-53105 Bonn, Germany</nlm:aff>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Institute for Medical Biometry, Informatics, and Epidemiology, University Hospital of Bonn, Sigmund-Freud-Str. 25, D-53105 Bonn</wicri:regionArea>
<wicri:noRegion>53105 Bonn</wicri:noRegion>
<wicri:noRegion>53105 Bonn</wicri:noRegion>
<wicri:noRegion>D-53105 Bonn</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Pfarr, Kenneth" sort="Pfarr, Kenneth" uniqKey="Pfarr K" first="Kenneth" last="Pfarr">Kenneth Pfarr</name>
<affiliation wicri:level="1"><nlm:aff id="I1">Institute of Medical Microbiology, Immunology and Parasitology, University Hospital of Bonn, Sigmund-Freud-Str. 25, D-53105 Bonn, Germany</nlm:aff>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Institute of Medical Microbiology, Immunology and Parasitology, University Hospital of Bonn, Sigmund-Freud-Str. 25, D-53105 Bonn</wicri:regionArea>
<wicri:noRegion>53105 Bonn</wicri:noRegion>
<wicri:noRegion>53105 Bonn</wicri:noRegion>
<wicri:noRegion>D-53105 Bonn</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Hoerauf, Achim" sort="Hoerauf, Achim" uniqKey="Hoerauf A" first="Achim" last="Hoerauf">Achim Hoerauf</name>
<affiliation wicri:level="1"><nlm:aff id="I1">Institute of Medical Microbiology, Immunology and Parasitology, University Hospital of Bonn, Sigmund-Freud-Str. 25, D-53105 Bonn, Germany</nlm:aff>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Institute of Medical Microbiology, Immunology and Parasitology, University Hospital of Bonn, Sigmund-Freud-Str. 25, D-53105 Bonn</wicri:regionArea>
<wicri:noRegion>53105 Bonn</wicri:noRegion>
<wicri:noRegion>53105 Bonn</wicri:noRegion>
<wicri:noRegion>D-53105 Bonn</wicri:noRegion>
</affiliation>
</author>
</analytic>
<series><title level="j">Parasites & Vectors</title>
<idno type="eISSN">1756-3305</idno>
<imprint><date when="2014">2014</date>
</imprint>
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<front><div type="abstract" xml:lang="en"><sec><title>Background</title>
<p>Lymphatic filariasis caused by <italic>Wuchereria bancrofti</italic>
or <italic>Brugia</italic>
spp. is a public health problem in developing countries. To monitor bancroftian filariasis infections, Circulating Filarial Antigen (CFA) test is commonly used, but for brugian infections only microfilariae (Mf) microscopy and indirect IgG4 antibody analyses are available. Improved diagnostics for detecting latent infections are required.</p>
</sec>
<sec><title>Methods</title>
<p>An optimized real-time PCR targeting the brugian <italic>HhaI</italic>
repeat was validated with plasma from microfilariae negative Mongolian gerbils (jirds) infected with <italic>B. malayi</italic>
. Plasma samples from microfilaremic patients infected with <italic>B. malayi</italic>
or <italic>W. bancrofti</italic>
were used as positive and negative controls, respectively. PCR results of plasma samples from a transmigrant population in a <italic>B. malayi</italic>
endemic area were compared to those of life-long residents in the same endemic area; and to IgG4 serology results from the same population. To discriminate between active infections and larval exposure a threshold was determined by correlation and Receiver-Operating Characteristics (ROC) curve analyses.</p>
</sec>
<sec><title>Results</title>
<p>The PCR detected <italic>HhaI</italic>
in pre-patent (56 dpi) <italic>B. malayi</italic>
infected jirds and <italic>B. malayi</italic>
Mf-positive patients from Central Sulawesi, Indonesia. <italic>HhaI</italic>
was also detected in 9/9 elephantiasis patients. In South Sulawesi 87.4% of the transmigrants and life-long residents (94% Mf-negative) were <italic>HhaI</italic>
PCR positive. Based on ROC-curve analysis a threshold for active infections was set to >53 <italic>HhaI</italic>
copies/μl (AUC: 0.854).</p>
</sec>
<sec><title>Conclusions</title>
<p>The results demonstrate that the <italic>HhaI</italic>
PCR detects brugian infections with greater sensitivity than the IgG4 test, most notably in Mf-negative patients (<italic>i.e</italic>
. pre-patent or latent infections).</p>
</sec>
</div>
</front>
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<pmc article-type="research-article" xml:lang="en"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Parasit Vectors</journal-id>
<journal-id journal-id-type="iso-abbrev">Parasit Vectors</journal-id>
<journal-title-group><journal-title>Parasites & Vectors</journal-title>
</journal-title-group>
<issn pub-type="epub">1756-3305</issn>
<publisher><publisher-name>BioMed Central</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">24685183</article-id>
<article-id pub-id-type="pmc">4021971</article-id>
<article-id pub-id-type="publisher-id">1756-3305-7-146</article-id>
<article-id pub-id-type="doi">10.1186/1756-3305-7-146</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Research</subject>
</subj-group>
</article-categories>
<title-group><article-title>Real-time PCR detection of the <italic>HhaI</italic>
tandem DNA repeat in pre- and post-patent <italic>Brugia malayi</italic>
infections: a study in Indonesian transmigrants</article-title>
</title-group>
<contrib-group><contrib contrib-type="author" id="A1"><name><surname>Albers</surname>
<given-names>Anna</given-names>
</name>
<xref ref-type="aff" rid="I1">1</xref>
<email>albers@microbiology-bonn.de</email>
</contrib>
<contrib contrib-type="author" id="A2"><name><surname>Sartono</surname>
<given-names>Erliyani</given-names>
</name>
<xref ref-type="aff" rid="I2">2</xref>
<email>E.Sartono@lumc.nl</email>
</contrib>
<contrib contrib-type="author" id="A3"><name><surname>Wahyuni</surname>
<given-names>Sitti</given-names>
</name>
<xref ref-type="aff" rid="I3">3</xref>
<email>wahyunim@indosat.net.id</email>
</contrib>
<contrib contrib-type="author" id="A4"><name><surname>Yazdanbakhsh</surname>
<given-names>Maria</given-names>
</name>
<xref ref-type="aff" rid="I2">2</xref>
<email>M.Yazdanbakhsh@lumc.nl</email>
</contrib>
<contrib contrib-type="author" id="A5"><name><surname>Maizels</surname>
<given-names>Rick M</given-names>
</name>
<xref ref-type="aff" rid="I4">4</xref>
<email>rick.maizels@ed.ac.uk</email>
</contrib>
<contrib contrib-type="author" id="A6"><name><surname>Klarmann-Schulz</surname>
<given-names>Ute</given-names>
</name>
<xref ref-type="aff" rid="I1">1</xref>
<xref ref-type="aff" rid="I5">5</xref>
<email>Ute.Klarmann@ukb.uni-bonn.de</email>
</contrib>
<contrib contrib-type="author" corresp="yes" id="A7"><name><surname>Pfarr</surname>
<given-names>Kenneth</given-names>
</name>
<xref ref-type="aff" rid="I1">1</xref>
<email>pfarr@microbiology-bonn.de</email>
</contrib>
<contrib contrib-type="author" id="A8"><name><surname>Hoerauf</surname>
<given-names>Achim</given-names>
</name>
<xref ref-type="aff" rid="I1">1</xref>
<email>hoerauf@microbiology-bonn.de</email>
</contrib>
</contrib-group>
<aff id="I1"><label>1</label>
Institute of Medical Microbiology, Immunology and Parasitology, University Hospital of Bonn, Sigmund-Freud-Str. 25, D-53105 Bonn, Germany</aff>
<aff id="I2"><label>2</label>
Department of Parasitology, Leiden University Medical Center, P.O Box 9600, 2300, RC Leiden, The Netherlands</aff>
<aff id="I3"><label>3</label>
Parasitology Department, Medical Faculty, Hasanuddin University, Jl. Perintis Kemerdekaan 10 Tamalanrea, Makassar 90245, Indonesia</aff>
<aff id="I4"><label>4</label>
Institute for Immunology & Infection Research, University of Edinburgh, Ashworth Laboratories, Edinburgh EH9 3JT, UK</aff>
<aff id="I5"><label>5</label>
Institute for Medical Biometry, Informatics, and Epidemiology, University Hospital of Bonn, Sigmund-Freud-Str. 25, D-53105 Bonn, Germany</aff>
<pub-date pub-type="collection"><year>2014</year>
</pub-date>
<pub-date pub-type="epub"><day>31</day>
<month>3</month>
<year>2014</year>
</pub-date>
<volume>7</volume>
<fpage>146</fpage>
<lpage>146</lpage>
<history><date date-type="received"><day>12</day>
<month>9</month>
<year>2013</year>
</date>
<date date-type="accepted"><day>12</day>
<month>3</month>
<year>2014</year>
</date>
</history>
<permissions><copyright-statement>Copyright © 2014 Albers et al.; licensee BioMed Central Ltd.</copyright-statement>
<copyright-year>2014</copyright-year>
<copyright-holder>Albers et al.; licensee BioMed Central Ltd.</copyright-holder>
<license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by/2.0"><license-p>This is an Open Access article distributed under the terms of the Creative Commons Attribution License (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/2.0">http://creativecommons.org/licenses/by/2.0</ext-link>
), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/publicdomain/zero/1.0/">http://creativecommons.org/publicdomain/zero/1.0/</ext-link>
) applies to the data made available in this article, unless otherwise stated.</license-p>
</license>
</permissions>
<self-uri xlink:href="http://www.parasitesandvectors.com/content/7/1/146"></self-uri>
<abstract><sec><title>Background</title>
<p>Lymphatic filariasis caused by <italic>Wuchereria bancrofti</italic>
or <italic>Brugia</italic>
spp. is a public health problem in developing countries. To monitor bancroftian filariasis infections, Circulating Filarial Antigen (CFA) test is commonly used, but for brugian infections only microfilariae (Mf) microscopy and indirect IgG4 antibody analyses are available. Improved diagnostics for detecting latent infections are required.</p>
</sec>
<sec><title>Methods</title>
<p>An optimized real-time PCR targeting the brugian <italic>HhaI</italic>
repeat was validated with plasma from microfilariae negative Mongolian gerbils (jirds) infected with <italic>B. malayi</italic>
. Plasma samples from microfilaremic patients infected with <italic>B. malayi</italic>
or <italic>W. bancrofti</italic>
were used as positive and negative controls, respectively. PCR results of plasma samples from a transmigrant population in a <italic>B. malayi</italic>
endemic area were compared to those of life-long residents in the same endemic area; and to IgG4 serology results from the same population. To discriminate between active infections and larval exposure a threshold was determined by correlation and Receiver-Operating Characteristics (ROC) curve analyses.</p>
</sec>
<sec><title>Results</title>
<p>The PCR detected <italic>HhaI</italic>
in pre-patent (56 dpi) <italic>B. malayi</italic>
infected jirds and <italic>B. malayi</italic>
Mf-positive patients from Central Sulawesi, Indonesia. <italic>HhaI</italic>
was also detected in 9/9 elephantiasis patients. In South Sulawesi 87.4% of the transmigrants and life-long residents (94% Mf-negative) were <italic>HhaI</italic>
PCR positive. Based on ROC-curve analysis a threshold for active infections was set to >53 <italic>HhaI</italic>
copies/μl (AUC: 0.854).</p>
</sec>
<sec><title>Conclusions</title>
<p>The results demonstrate that the <italic>HhaI</italic>
PCR detects brugian infections with greater sensitivity than the IgG4 test, most notably in Mf-negative patients (<italic>i.e</italic>
. pre-patent or latent infections).</p>
</sec>
</abstract>
<kwd-group><kwd><italic>Brugia malayi</italic>
</kwd>
<kwd>amicrofilaremic</kwd>
<kwd>real-time PCR</kwd>
<kwd><italic>HhaI</italic>
</kwd>
<kwd>pre-patent</kwd>
<kwd>detection</kwd>
</kwd-group>
</article-meta>
</front>
</pmc>
<affiliations><list><country><li>Allemagne</li>
<li>Indonésie</li>
<li>Pays-Bas</li>
<li>Royaume-Uni</li>
</country>
<region><li>Écosse</li>
</region>
<settlement><li>Édimbourg</li>
</settlement>
<orgName><li>Université d'Édimbourg</li>
</orgName>
</list>
<tree><country name="Allemagne"><noRegion><name sortKey="Albers, Anna" sort="Albers, Anna" uniqKey="Albers A" first="Anna" last="Albers">Anna Albers</name>
</noRegion>
<name sortKey="Hoerauf, Achim" sort="Hoerauf, Achim" uniqKey="Hoerauf A" first="Achim" last="Hoerauf">Achim Hoerauf</name>
<name sortKey="Klarmann Schulz, Ute" sort="Klarmann Schulz, Ute" uniqKey="Klarmann Schulz U" first="Ute" last="Klarmann-Schulz">Ute Klarmann-Schulz</name>
<name sortKey="Klarmann Schulz, Ute" sort="Klarmann Schulz, Ute" uniqKey="Klarmann Schulz U" first="Ute" last="Klarmann-Schulz">Ute Klarmann-Schulz</name>
<name sortKey="Pfarr, Kenneth" sort="Pfarr, Kenneth" uniqKey="Pfarr K" first="Kenneth" last="Pfarr">Kenneth Pfarr</name>
</country>
<country name="Pays-Bas"><noRegion><name sortKey="Sartono, Erliyani" sort="Sartono, Erliyani" uniqKey="Sartono E" first="Erliyani" last="Sartono">Erliyani Sartono</name>
</noRegion>
<name sortKey="Yazdanbakhsh, Maria" sort="Yazdanbakhsh, Maria" uniqKey="Yazdanbakhsh M" first="Maria" last="Yazdanbakhsh">Maria Yazdanbakhsh</name>
</country>
<country name="Indonésie"><noRegion><name sortKey="Wahyuni, Sitti" sort="Wahyuni, Sitti" uniqKey="Wahyuni S" first="Sitti" last="Wahyuni">Sitti Wahyuni</name>
</noRegion>
</country>
<country name="Royaume-Uni"><region name="Écosse"><name sortKey="Maizels, Rick M" sort="Maizels, Rick M" uniqKey="Maizels R" first="Rick M" last="Maizels">Rick M. Maizels</name>
</region>
</country>
</tree>
</affiliations>
</record>
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