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Evaluation of PCR-ELISA as a tool for monitoring transmission of Wuchereria bancrofti in District of Gampaha, Sri Lanka

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Evaluation of PCR-ELISA as a tool for monitoring transmission of Wuchereria bancrofti in District of Gampaha, Sri Lanka

Auteurs : Asha Dilrukshi Wijegunawardana [Sri Lanka] ; Nilmini Silva Gunawardane [Sri Lanka] ; Chanditha Hapuarachchi [Singapour] ; Aresha Manamperi [Sri Lanka] ; Kithsiri Gunawardena [Sri Lanka] ; Wimaladharma Abeyewickrama [Sri Lanka]

Source :

Asian Pacific Journal of Tropical Biomedicine [ 2221-1691 ] ; 2013.

RBID : PMC:3642448

Abstract

Objective

To compare Wuchereria bancrofti (W. bancrofti) infection rates of Culex quinquefasciatus, using dissection and PCR-ELISA in two consecutive time periods (from 2007 to 2008 and from 2008 to 2009).

Methods

Mosquitoes were collected in 30 sentinel and 15 non-sentinel sites in 15 Medical Officer of Health areas of Gampaha District known for the presence of W. bancrofti transmission in two consecutive time period of 2007 to 2008 and 2008 to 2009. Captured mosquitoes were dissected to determine the W. bancrofti larvae (L₁, L₂, L₃). PCR was carried out using DNA extracted from mosquito pools (15 body parts/pool) utilizing the primers specific for Wb-SspI repeat. PCR products were analyzed by hybridization ELISA using fluorescein-labeled wild type specific probes. The prevalence of infected/infective mosquitoes in PCR pools (3 pools/site) was estimated using the PoolScreen™ algorithm and a novel probability-based method.

Results

Of 45 batches of mosquitoes dissected, W. bancrofti infected mosquitoes were found in 19 and 13 batches, with an infection rate of 13.29% and 3.10% with mean larval density of 8.7 and 1.0 larvae per mosquito for two study periods in the Gampaha District. Total of 405 pools of head, thorax and abdomen were processed by PCR-ELISA for each year. Of these, 51 and 31 pools were positive for W. bancrofti in the two study periods respectively. The association of dissection based prevalence rates with PCR based rates as determined by the Pearson correlation coefficient were 0.176 and 0.890 respectively for the two periods.

Conclusions

Data indicate that PCR-ELISA is more sensitive than the traditional dissection techniques for monitoring transmission intensity.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3642448

DOI: 10.1016/S2221-1691(13)60081-7
PubMed: 23646302
PubMed Central: 3642448

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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Evaluation of PCR-ELISA as a tool for monitoring transmission of <italic>Wuchereria bancrofti</italic>
 in District of Gampaha, Sri Lanka</title>
<author><name sortKey="Wijegunawardana, Asha Dilrukshi" sort="Wijegunawardana, Asha Dilrukshi" uniqKey="Wijegunawardana A" first="Asha Dilrukshi" last="Wijegunawardana">Asha Dilrukshi Wijegunawardana</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><addr-line>Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya, Ragama, Sri Lanka</addr-line>
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<country xml:lang="fr">Sri Lanka</country>
<wicri:regionArea>Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya, Ragama</wicri:regionArea>
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<author><name sortKey="Gunawardane, Nilmini Silva" sort="Gunawardane, Nilmini Silva" uniqKey="Gunawardane N" first="Nilmini Silva" last="Gunawardane">Nilmini Silva Gunawardane</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><addr-line>Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya, Ragama, Sri Lanka</addr-line>
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<country xml:lang="fr">Sri Lanka</country>
<wicri:regionArea>Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya, Ragama</wicri:regionArea>
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<author><name sortKey="Hapuarachchi, Chanditha" sort="Hapuarachchi, Chanditha" uniqKey="Hapuarachchi C" first="Chanditha" last="Hapuarachchi">Chanditha Hapuarachchi</name>
<affiliation wicri:level="1"><nlm:aff id="aff3"><addr-line>Environmental Authority, Singapore</addr-line>
</nlm:aff>
<country xml:lang="fr">Singapour</country>
<wicri:regionArea>Environmental Authority</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Manamperi, Aresha" sort="Manamperi, Aresha" uniqKey="Manamperi A" first="Aresha" last="Manamperi">Aresha Manamperi</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><addr-line>Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya, Ragama, Sri Lanka</addr-line>
</nlm:aff>
<country xml:lang="fr">Sri Lanka</country>
<wicri:regionArea>Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya, Ragama</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Gunawardena, Kithsiri" sort="Gunawardena, Kithsiri" uniqKey="Gunawardena K" first="Kithsiri" last="Gunawardena">Kithsiri Gunawardena</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><addr-line>Department of Parasitology, Faculty of Medicine, University of Kelaniya, Ragama, Sri Lanka</addr-line>
</nlm:aff>
<country xml:lang="fr">Sri Lanka</country>
<wicri:regionArea>Department of Parasitology, Faculty of Medicine, University of Kelaniya, Ragama</wicri:regionArea>
</affiliation>
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<author><name sortKey="Abeyewickrama, Wimaladharma" sort="Abeyewickrama, Wimaladharma" uniqKey="Abeyewickrama W" first="Wimaladharma" last="Abeyewickrama">Wimaladharma Abeyewickrama</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><addr-line>Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya, Ragama, Sri Lanka</addr-line>
</nlm:aff>
<country xml:lang="fr">Sri Lanka</country>
<wicri:regionArea>Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya, Ragama</wicri:regionArea>
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</nlm:aff>
<country xml:lang="fr">Sri Lanka</country>
<wicri:regionArea>Department of Parasitology, Faculty of Medicine, University of Kelaniya, Ragama</wicri:regionArea>
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<series><title level="j">Asian Pacific Journal of Tropical Biomedicine</title>
<idno type="ISSN">2221-1691</idno>
<imprint><date when="2013">2013</date>
</imprint>
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<front><div type="abstract" xml:lang="en"><sec><title>Objective</title>
<p>To compare <italic>Wuchereria bancrofti</italic>
 (<italic>W. bancrofti</italic>
) infection rates of <italic>Culex quinquefasciatus</italic>
, using dissection and PCR-ELISA in two consecutive time periods (from 2007 to 2008 and from 2008 to 2009).</p>
</sec>
<sec><title>Methods</title>
<p>Mosquitoes were collected in 30 sentinel and 15 non-sentinel sites in 15 Medical Officer of Health areas of Gampaha District known for the presence of <italic>W. bancrofti</italic>
 transmission in two consecutive time period of 2007 to 2008 and 2008 to 2009. Captured mosquitoes were dissected to determine the <italic>W. bancrofti</italic>
 larvae (L<sub>1</sub>
, L<sub>2</sub>
, L<sub>3</sub>
). PCR was carried out using DNA extracted from mosquito pools (15 body parts/pool) utilizing the primers specific for Wb-SspI repeat. PCR products were analyzed by hybridization ELISA using fluorescein-labeled wild type specific probes. The prevalence of infected/infective mosquitoes in PCR pools (3 pools/site) was estimated using the PoolScreen™ algorithm and a novel probability-based method.</p>
</sec>
<sec><title>Results</title>
<p>Of 45 batches of mosquitoes dissected, <italic>W. bancrofti</italic>
 infected mosquitoes were found in 19 and 13 batches, with an infection rate of 13.29% and 3.10% with mean larval density of 8.7 and 1.0 larvae per mosquito for two study periods in the Gampaha District. Total of 405 pools of head, thorax and abdomen were processed by PCR-ELISA for each year. Of these, 51 and 31 pools were positive for <italic>W. bancrofti</italic>
 in the two study periods respectively. The association of dissection based prevalence rates with PCR based rates as determined by the Pearson correlation coefficient were 0.176 and 0.890 respectively for the two periods.</p>
</sec>
<sec><title>Conclusions</title>
<p>Data indicate that PCR-ELISA is more sensitive than the traditional dissection techniques for monitoring transmission intensity.</p>
</sec>
</div>
</front>
</TEI>
<pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
  <front><journal-meta><journal-id journal-id-type="nlm-ta">Asian Pac J Trop Biomed</journal-id>
<journal-id journal-id-type="iso-abbrev">Asian Pac J Trop Biomed</journal-id>
<journal-id journal-id-type="publisher-id">APJTB</journal-id>
<journal-title-group><journal-title>Asian Pacific Journal of Tropical Biomedicine</journal-title>
</journal-title-group>
<issn pub-type="ppub">2221-1691</issn>
<publisher><publisher-name>Asian Pacific Tropical Medicine Press</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">23646302</article-id>
<article-id pub-id-type="pmc">3642448</article-id>
<article-id pub-id-type="publisher-id">apjtb-03-05-381</article-id>
<article-id pub-id-type="doi">10.1016/S2221-1691(13)60081-7</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Basic Researches</subject>
</subj-group>
</article-categories>
<title-group><article-title>Evaluation of PCR-ELISA as a tool for monitoring transmission of <italic>Wuchereria bancrofti</italic>
 in District of Gampaha, Sri Lanka</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Wijegunawardana</surname>
<given-names>Asha Dilrukshi</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Gunawardane</surname>
<given-names>Nilmini Silva</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="corresp" rid="cor1">*</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Hapuarachchi</surname>
<given-names>Chanditha</given-names>
</name>
<xref ref-type="aff" rid="aff3">3</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Manamperi</surname>
<given-names>Aresha</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Gunawardena</surname>
<given-names>Kithsiri</given-names>
</name>
<xref ref-type="aff" rid="aff2">2</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Abeyewickrama</surname>
<given-names>Wimaladharma</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="aff" rid="aff2">2</xref>
</contrib>
</contrib-group>
<aff id="aff1"><label>1</label>
<addr-line>Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya, Ragama, Sri Lanka</addr-line>
</aff>
<aff id="aff2"><label>2</label>
<addr-line>Department of Parasitology, Faculty of Medicine, University of Kelaniya, Ragama, Sri Lanka</addr-line>
</aff>
<aff id="aff3"><label>3</label>
<addr-line>Environmental Authority, Singapore</addr-line>
</aff>
<contrib-group><contrib contrib-type="reviewer"><name><surname>Latif</surname>
<given-names>Baha</given-names>
</name>
</contrib>
</contrib-group>
<aff id="aff4"><addr-line>Professor, Faculty of Medicine, Universiti Teknologi MARA, Sungai Buloh, Selangor, Malaysia</addr-line>
 Tel:<phone>+6 0173145649</phone>
 Fax:<fax>+6 0361267073</fax>
 E-mail:<email>bahalatif@yahoo.com</email>
</aff>
<author-notes><corresp id="cor1">*Corresponding author: Nilmini Silva Gunawardane, Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya, Ragama, Sri Lanka. Tel: <phone>011-2960483</phone>
 Fax: <fax>011-2958337</fax>
 E-mail: <email>nilminis@graduate.hku.hk</email>
</corresp>
</author-notes>
<pub-date pub-type="ppub"><month>5</month>
<year>2013</year>
</pub-date>
<volume>3</volume>
<issue>5</issue>
<fpage>381</fpage>
<lpage>387</lpage>
<history><date date-type="received"><day>4</day>
<month>3</month>
<year>2013</year>
</date>
<date date-type="accepted"><day>5</day>
<month>5</month>
<year>2013</year>
</date>
</history>
<permissions><copyright-statement>© 2013 by the Asian Pacific Journal of Tropical Biomedicine. All rights reserved.</copyright-statement>
<copyright-year>2013</copyright-year>
</permissions>
<abstract><sec><title>Objective</title>
<p>To compare <italic>Wuchereria bancrofti</italic>
 (<italic>W. bancrofti</italic>
) infection rates of <italic>Culex quinquefasciatus</italic>
, using dissection and PCR-ELISA in two consecutive time periods (from 2007 to 2008 and from 2008 to 2009).</p>
</sec>
<sec><title>Methods</title>
<p>Mosquitoes were collected in 30 sentinel and 15 non-sentinel sites in 15 Medical Officer of Health areas of Gampaha District known for the presence of <italic>W. bancrofti</italic>
 transmission in two consecutive time period of 2007 to 2008 and 2008 to 2009. Captured mosquitoes were dissected to determine the <italic>W. bancrofti</italic>
 larvae (L<sub>1</sub>
, L<sub>2</sub>
, L<sub>3</sub>
). PCR was carried out using DNA extracted from mosquito pools (15 body parts/pool) utilizing the primers specific for Wb-SspI repeat. PCR products were analyzed by hybridization ELISA using fluorescein-labeled wild type specific probes. The prevalence of infected/infective mosquitoes in PCR pools (3 pools/site) was estimated using the PoolScreen™ algorithm and a novel probability-based method.</p>
</sec>
<sec><title>Results</title>
<p>Of 45 batches of mosquitoes dissected, <italic>W. bancrofti</italic>
 infected mosquitoes were found in 19 and 13 batches, with an infection rate of 13.29% and 3.10% with mean larval density of 8.7 and 1.0 larvae per mosquito for two study periods in the Gampaha District. Total of 405 pools of head, thorax and abdomen were processed by PCR-ELISA for each year. Of these, 51 and 31 pools were positive for <italic>W. bancrofti</italic>
 in the two study periods respectively. The association of dissection based prevalence rates with PCR based rates as determined by the Pearson correlation coefficient were 0.176 and 0.890 respectively for the two periods.</p>
</sec>
<sec><title>Conclusions</title>
<p>Data indicate that PCR-ELISA is more sensitive than the traditional dissection techniques for monitoring transmission intensity.</p>
</sec>
</abstract>
<kwd-group><kwd><italic>Wuchereria bancrofti</italic>
</kwd>
<kwd><italic>Culex quinquefasciatus</italic>
</kwd>
<kwd>Dissection</kwd>
<kwd>PCR-ELISA</kwd>
<kwd>Sri Lanka</kwd>
</kwd-group>
</article-meta>
</front>
</pmc>
</record>

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Evaluation of PCR-ELISA as a tool for monitoring transmission of Wuchereria bancrofti in District of Gampaha, Sri Lanka

Evaluation of PCR-ELISA as a tool for monitoring transmission of Wuchereria bancrofti in District of Gampaha, Sri Lanka

Source :

Abstract

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