Diagnosis of Brugian Filariasis by Loop-Mediated Isothermal Amplification
Identifieur interne : 004328 ( Pmc/Curation ); précédent : 004327; suivant : 004329Diagnosis of Brugian Filariasis by Loop-Mediated Isothermal Amplification
Auteurs : Catherine B. Poole ; Nathan A. Tanner ; Yinhua Zhang ; Thomas C. Evans ; Clotilde K. S. CarlowSource :
- PLoS Neglected Tropical Diseases [ 1935-2727 ] ; 2012.
Abstract
In this study we developed and evaluated a
Url:
DOI: 10.1371/journal.pntd.0001948
PubMed: 23272258
PubMed Central: 3521703
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Diagnosis of Brugian Filariasis by Loop-Mediated Isothermal Amplification</title><author><name sortKey="Poole, Catherine B" sort="Poole, Catherine B" uniqKey="Poole C" first="Catherine B." last="Poole">Catherine B. Poole</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Tanner, Nathan A" sort="Tanner, Nathan A" uniqKey="Tanner N" first="Nathan A." last="Tanner">Nathan A. Tanner</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Zhang, Yinhua" sort="Zhang, Yinhua" uniqKey="Zhang Y" first="Yinhua" last="Zhang">Yinhua Zhang</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Evans, Thomas C" sort="Evans, Thomas C" uniqKey="Evans T" first="Thomas C." last="Evans">Thomas C. Evans</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Carlow, Clotilde K S" sort="Carlow, Clotilde K S" uniqKey="Carlow C" first="Clotilde K. S." last="Carlow">Clotilde K. S. Carlow</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">23272258</idno><idno type="pmc">3521703</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3521703</idno><idno type="RBID">PMC:3521703</idno><idno type="doi">10.1371/journal.pntd.0001948</idno><date when="2012">2012</date><idno type="wicri:Area/Pmc/Corpus">004329</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">004329</idno><idno type="wicri:Area/Pmc/Curation">004328</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">004328</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Diagnosis of Brugian Filariasis by Loop-Mediated Isothermal Amplification</title><author><name sortKey="Poole, Catherine B" sort="Poole, Catherine B" uniqKey="Poole C" first="Catherine B." last="Poole">Catherine B. Poole</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Tanner, Nathan A" sort="Tanner, Nathan A" uniqKey="Tanner N" first="Nathan A." last="Tanner">Nathan A. Tanner</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Zhang, Yinhua" sort="Zhang, Yinhua" uniqKey="Zhang Y" first="Yinhua" last="Zhang">Yinhua Zhang</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Evans, Thomas C" sort="Evans, Thomas C" uniqKey="Evans T" first="Thomas C." last="Evans">Thomas C. Evans</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Carlow, Clotilde K S" sort="Carlow, Clotilde K S" uniqKey="Carlow C" first="Clotilde K. S." last="Carlow">Clotilde K. S. Carlow</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">PLoS Neglected Tropical Diseases</title><idno type="ISSN">1935-2727</idno><idno type="eISSN">1935-2735</idno><imprint><date when="2012">2012</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>In this study we developed and evaluated a <italic>Brugia Hha</italic> I repeat loop-mediated isothermal amplification (LAMP) assay for the rapid detection of <italic>Brugia</italic> genomic DNA. Amplification was detected using turbidity or fluorescence as readouts. Reactions generated a turbidity threshold value or a clear visual positive within 30 minutes using purified genomic DNA equivalent to one microfilaria. Similar results were obtained using DNA isolated from blood samples containing <italic>B. malayi</italic> microfilariae. Amplification was specific to <italic>B. malayi</italic> and <italic>B. timori</italic>, as no turbidity was observed using DNA from the related filarial parasites <italic>Wuchereria bancrofti</italic>, <italic>Onchocerca volvulus</italic> or <italic>Dirofilaria immitis</italic>, or from human or mosquito. Furthermore, the assay was most robust using a new strand-displacing DNA polymerase termed <italic>Bst 2.0</italic> compared to wild-type <italic>Bst</italic> DNA polymerase, large fragment. The results indicate that the <italic>Brugia Hha</italic> I repeat LAMP assay is rapid, sensitive and <italic>Brugia</italic>-specific with the potential to be developed further as a field tool for diagnosis and mapping of brugian filariasis.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Ottesen, Ea" uniqKey="Ottesen E">EA Ottesen</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Weil, Gj" uniqKey="Weil G">GJ Weil</name></author><author><name sortKey="Lammie, Pj" uniqKey="Lammie P">PJ Lammie</name></author><author><name sortKey="Weiss, N" uniqKey="Weiss N">N Weiss</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mccarthy, Js" uniqKey="Mccarthy J">JS McCarthy</name></author><author><name sortKey="Lustigman, S" uniqKey="Lustigman S">S Lustigman</name></author><author><name sortKey="Yang, Gj" uniqKey="Yang G">GJ 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article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">PLoS Negl Trop Dis</journal-id><journal-id journal-id-type="iso-abbrev">PLoS Negl Trop Dis</journal-id><journal-id journal-id-type="publisher-id">plos</journal-id><journal-id journal-id-type="pmc">plosntds</journal-id><journal-title-group><journal-title>PLoS Neglected Tropical Diseases</journal-title></journal-title-group><issn pub-type="ppub">1935-2727</issn><issn pub-type="epub">1935-2735</issn><publisher><publisher-name>Public Library of Science</publisher-name><publisher-loc>San Francisco, USA</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">23272258</article-id><article-id pub-id-type="pmc">3521703</article-id><article-id pub-id-type="publisher-id">PNTD-D-12-00911</article-id><article-id pub-id-type="doi">10.1371/journal.pntd.0001948</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group><subj-group subj-group-type="Discipline-v2"><subject>Medicine</subject><subj-group><subject>Diagnostic Medicine</subject></subj-group><subj-group><subject>Infectious Diseases</subject><subj-group><subject>Neglected Tropical Diseases</subject><subject>Parasitic Diseases</subject></subj-group></subj-group></subj-group></article-categories><title-group><article-title>Diagnosis of Brugian Filariasis by Loop-Mediated Isothermal Amplification</article-title><alt-title alt-title-type="running-head">Detection of Brugian Filariasis Using LAMP</alt-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Poole</surname><given-names>Catherine B.</given-names></name><xref ref-type="aff" rid="aff1"></xref></contrib><contrib contrib-type="author"><name><surname>Tanner</surname><given-names>Nathan A.</given-names></name><xref ref-type="aff" rid="aff1"></xref></contrib><contrib contrib-type="author"><name><surname>Zhang</surname><given-names>Yinhua</given-names></name><xref ref-type="aff" rid="aff1"></xref></contrib><contrib contrib-type="author"><name><surname>Evans</surname><given-names>Thomas C.</given-names><suffix>Jr.</suffix></name><xref ref-type="aff" rid="aff1"></xref></contrib><contrib contrib-type="author"><name><surname>Carlow</surname><given-names>Clotilde K. S.</given-names></name><xref ref-type="aff" rid="aff1"></xref><xref ref-type="corresp" rid="cor1"><sup>*</sup></xref></contrib></contrib-group><aff id="aff1"><addr-line>New England Biolabs, Ipswich, Massachusetts, United States of America</addr-line></aff><contrib-group><contrib contrib-type="editor"><name><surname>McCarthy</surname><given-names>James S.</given-names></name><role>Editor</role><xref ref-type="aff" rid="edit1"></xref></contrib></contrib-group><aff id="edit1"><addr-line>Queensland Institute for Medical Research, Australia</addr-line></aff><author-notes><corresp id="cor1">* E-mail: <email>carlow@neb.com</email></corresp><fn fn-type="conflict"><p>CBP, NAT, YZ, TCE and CKSC have read the journal's policy and have the following conflicts: CBP, NAT, YZ, TCE and CKSC are employees of New England Biolabs. The author's affiliation with the funder does not alter adherence to all the PLOS Neglected Tropical Diseases policies on sharing data and materials.</p></fn><fn fn-type="con"><p>Conceived and designed the experiments: CBP NAT YZ TCE CKSC. Performed the experiments: CBP. Analyzed the data: CBP NAT YZ TCE CKSC. Contributed reagents/materials/analysis tools: CBP NAT YZ TCE. Wrote the paper: CBP NAT YZ TCE CKSC.</p></fn></author-notes><pub-date pub-type="collection"><month>12</month><year>2012</year></pub-date><pub-date pub-type="epub"><day>13</day><month>12</month><year>2012</year></pub-date><volume>6</volume><issue>12</issue><elocation-id>e1948</elocation-id><history><date date-type="received"><day>25</day><month>7</month><year>2012</year></date><date date-type="accepted"><day>25</day><month>10</month><year>2012</year></date></history><permissions><copyright-year>2012</copyright-year><copyright-holder>Poole et al</copyright-holder><license><license-p>This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</license-p></license></permissions><abstract><p>In this study we developed and evaluated a <italic>Brugia Hha</italic> I repeat loop-mediated isothermal amplification (LAMP) assay for the rapid detection of <italic>Brugia</italic> genomic DNA. Amplification was detected using turbidity or fluorescence as readouts. Reactions generated a turbidity threshold value or a clear visual positive within 30 minutes using purified genomic DNA equivalent to one microfilaria. Similar results were obtained using DNA isolated from blood samples containing <italic>B. malayi</italic> microfilariae. Amplification was specific to <italic>B. malayi</italic> and <italic>B. timori</italic>, as no turbidity was observed using DNA from the related filarial parasites <italic>Wuchereria bancrofti</italic>, <italic>Onchocerca volvulus</italic> or <italic>Dirofilaria immitis</italic>, or from human or mosquito. Furthermore, the assay was most robust using a new strand-displacing DNA polymerase termed <italic>Bst 2.0</italic> compared to wild-type <italic>Bst</italic> DNA polymerase, large fragment. The results indicate that the <italic>Brugia Hha</italic> I repeat LAMP assay is rapid, sensitive and <italic>Brugia</italic>-specific with the potential to be developed further as a field tool for diagnosis and mapping of brugian filariasis.</p></abstract><abstract abstract-type="summary"><title>Author Summary</title><p>Brugian filariasis is a debilitating neglected tropical disease caused by infection with the filarial parasites <italic>Brugia malayi</italic> or <italic>Brugia timori</italic>. Adult worms live in the lymphatic system and produce large numbers of microfilariae that predominantly circulate in the blood at night. Bloodsucking mosquitoes spread the disease by ingesting microfilariae that develop into infective stage larvae in the insect. In rural areas, diagnosis still relies largely on microscopic examination of night blood and morphological assessment of stained microfilariae. Loop-mediated isothermal amplification (LAMP) is a technique that can amplify DNA with high specificity, sensitivity and rapidity under isothermal conditions. The operational simplicity, versatility and low-cost of the technique make it particularly appealing for use in diagnosis and geographical mapping of neglected tropical diseases. In the present study, we have developed and evaluated a <italic>Brugia Hha</italic> I repeat LAMP assay for the rapid detection of <italic>B. malayi</italic> and <italic>B. timori</italic> genomic DNA. The results indicate that the <italic>Brugia Hha</italic> I repeat LAMP diagnostic assay is sensitive and rapid, detecting a single microfilariae in blood within 30 minutes, and <italic>Brugia-</italic>specific. The test has the potential to be developed further as a field tool for use in the implementation and management of mass drug administration programs for brugian filariasis.</p></abstract><funding-group><funding-statement>This work was funded by New England Biolabs. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.</funding-statement></funding-group><counts><page-count count="9"></page-count></counts></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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