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Detection of Brugia Parasite DNA in Human Blood by Real-Time PCR▿

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Detection of Brugia Parasite DNA in Human Blood by Real-Time PCR▿

Auteurs : Ramakrishna U. Rao ; Gary J. Weil ; Kerstin Fischer ; Taniawati Supali ; Peter Fischer

Source :

Journal of Clinical Microbiology [ 0095-1137 ] ; 2006.

RBID : PMC:1698366

Abstract

Brugian filariasis (caused by the nematodes Brugia malayi and B. timori) is an important cause of disability in Southeast Asia. Improved diagnostic tests are needed for filariasis elimination programs (to identify areas of endemicity and to monitor progress) and for diagnosis of the disease in infected individuals. We have developed and evaluated two real-time PCR assays for detecting Brugia DNA in human blood and compared the results of these assays to those of “gold standard” assays. One assay uses a TaqMan probe (TaqM) to amplifiy a 320-bp “HhaI repeat” DNA sequence. The other assay uses a minor groove binding probe (MGB) and modified nucleotides in primers (Eclipse MGB) to amplify a 120-bp fragment of the HhaI repeat. This assay detects 22 copies of the target sequence, and it is more sensitive than the TaqM assay. Both assays were evaluated with human blood samples from two different areas of endemicity. The MGB assay was as sensitive as membrane filtration and microscopy for the detection of B. malayi infection in 57 blood samples recovered at night from patients in Sulawesi, Indonesia. The MGB assay also detected parasite DNA in 17 of 31 (55%) of microfilaria-negative day blood samples from these subjects. This test was more sensitive than the conventional and the TaqM PCRs (and was almost as sensitive as night blood membrane filtration) for the detection of infection in 52 blood samples recovered at night from individuals in an area of B. timori endemicity on Alor Island, Indonesia, where microfilaria-positive individuals had low densities after mass treatment. Thus, the Eclipse MGB real-time PCR assay is a sensitive means of detecting Brugia parasite DNA in human blood.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1698366

DOI: 10.1128/JCM.00969-06
PubMed: 16957038
PubMed Central: 1698366

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PMC:1698366

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 </title>
<author><name sortKey="Rao, Ramakrishna U" sort="Rao, Ramakrishna U" uniqKey="Rao R" first="Ramakrishna U." last="Rao">Ramakrishna U. Rao</name>
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<series><title level="j">Journal of Clinical Microbiology</title>
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<front><div type="abstract" xml:lang="en"><p>Brugian filariasis (caused by the nematodes <italic>Brugia malayi</italic>
 and <italic>B. timori</italic>
) is an important cause of disability in Southeast Asia. Improved diagnostic tests are needed for filariasis elimination programs (to identify areas of endemicity and to monitor progress) and for diagnosis of the disease in infected individuals. We have developed and evaluated two real-time PCR assays for detecting <italic>Brugia</italic>
 DNA in human blood and compared the results of these assays to those of “gold standard” assays. One assay uses a TaqMan probe (TaqM) to amplifiy a 320-bp “HhaI repeat” DNA sequence. The other assay uses a minor groove binding probe (MGB) and modified nucleotides in primers (Eclipse MGB) to amplify a 120-bp fragment of the HhaI repeat. This assay detects 22 copies of the target sequence, and it is more sensitive than the TaqM assay. Both assays were evaluated with human blood samples from two different areas of endemicity. The MGB assay was as sensitive as membrane filtration and microscopy for the detection of <italic>B. malayi</italic>
 infection in 57 blood samples recovered at night from patients in Sulawesi, Indonesia. The MGB assay also detected parasite DNA in 17 of 31 (55%) of microfilaria-negative day blood samples from these subjects. This test was more sensitive than the conventional and the TaqM PCRs (and was almost as sensitive as night blood membrane filtration) for the detection of infection in 52 blood samples recovered at night from individuals in an area of <italic>B. timori</italic>
 endemicity on Alor Island, Indonesia, where microfilaria-positive individuals had low densities after mass treatment. Thus, the Eclipse MGB real-time PCR assay is a sensitive means of detecting <italic>Brugia</italic>
 parasite DNA in human blood.</p>
</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">J Clin Microbiol</journal-id>
<journal-id journal-id-type="publisher-id">jcm</journal-id>
<journal-title>Journal of Clinical Microbiology</journal-title>
<issn pub-type="ppub">0095-1137</issn>
<issn pub-type="epub">1098-660X</issn>
<publisher><publisher-name>American Society for Microbiology</publisher-name>
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<article-meta><article-id pub-id-type="pmid">16957038</article-id>
<article-id pub-id-type="pmc">1698366</article-id>
<article-id pub-id-type="publisher-id">0969-06</article-id>
<article-id pub-id-type="doi">10.1128/JCM.00969-06</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Parasitology</subject>
</subj-group>
</article-categories>
<title-group><article-title>Detection of <italic>Brugia</italic>
 Parasite DNA in Human Blood by Real-Time PCR<xref ref-type="fn" rid="fn1">▿</xref>
 </article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Rao</surname>
<given-names>Ramakrishna U.</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Weil</surname>
<given-names>Gary J.</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Fischer</surname>
<given-names>Kerstin</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Supali</surname>
<given-names>Taniawati</given-names>
</name>
<xref ref-type="aff" rid="aff1">2</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Fischer</surname>
<given-names>Peter</given-names>
</name>
<xref ref-type="aff" rid="aff1">1</xref>
<xref ref-type="corresp" rid="cor1">*</xref>
</contrib>
</contrib-group>
<aff id="aff1">Department of Internal Medicine, Infectious Diseases Division, Washington University School of Medicine, St. Louis, Missouri,<label>1</label>
 Department of Parasitology, Faculty of Medicine, University of Indonesia, Jakarta, Indonesia<label>2</label>
</aff>
<author-notes><fn id="cor1"><label>*</label>
<p>Corresponding author. Mailing address: Department of Internal Medicine, Infectious Diseases Division, Washington University School of Medicine, 660 South Euclid Avenue, Campus Box 8051, St. Louis, MO 63110. Phone: (314) 454-7876. Fax: (314) 454-5293. E-mail: <email>Pufische@im.wustl.edu</email>
.</p>
</fn>
</author-notes>
<pub-date pub-type="ppub"><month>11</month>
<year>2006</year>
</pub-date>
<pub-date pub-type="epub"><day>6</day>
<month>9</month>
<year>2006</year>
</pub-date>
<volume>44</volume>
<issue>11</issue>
<fpage>3887</fpage>
<lpage>3893</lpage>
<history><date date-type="received"><day>9</day>
<month>5</month>
<year>2006</year>
</date>
<date date-type="rev-recd"><day>6</day>
<month>7</month>
<year>2006</year>
</date>
<date date-type="accepted"><day>28</day>
<month>8</month>
<year>2006</year>
</date>
</history>
<copyright-statement>Copyright © 2006, American Society for Microbiology</copyright-statement>
<copyright-year>2006</copyright-year>
<self-uri xlink:title="pdf" xlink:href="zjm01106003887.pdf"></self-uri>
<abstract><p>Brugian filariasis (caused by the nematodes <italic>Brugia malayi</italic>
 and <italic>B. timori</italic>
) is an important cause of disability in Southeast Asia. Improved diagnostic tests are needed for filariasis elimination programs (to identify areas of endemicity and to monitor progress) and for diagnosis of the disease in infected individuals. We have developed and evaluated two real-time PCR assays for detecting <italic>Brugia</italic>
 DNA in human blood and compared the results of these assays to those of “gold standard” assays. One assay uses a TaqMan probe (TaqM) to amplifiy a 320-bp “HhaI repeat” DNA sequence. The other assay uses a minor groove binding probe (MGB) and modified nucleotides in primers (Eclipse MGB) to amplify a 120-bp fragment of the HhaI repeat. This assay detects 22 copies of the target sequence, and it is more sensitive than the TaqM assay. Both assays were evaluated with human blood samples from two different areas of endemicity. The MGB assay was as sensitive as membrane filtration and microscopy for the detection of <italic>B. malayi</italic>
 infection in 57 blood samples recovered at night from patients in Sulawesi, Indonesia. The MGB assay also detected parasite DNA in 17 of 31 (55%) of microfilaria-negative day blood samples from these subjects. This test was more sensitive than the conventional and the TaqM PCRs (and was almost as sensitive as night blood membrane filtration) for the detection of infection in 52 blood samples recovered at night from individuals in an area of <italic>B. timori</italic>
 endemicity on Alor Island, Indonesia, where microfilaria-positive individuals had low densities after mass treatment. Thus, the Eclipse MGB real-time PCR assay is a sensitive means of detecting <italic>Brugia</italic>
 parasite DNA in human blood.</p>
</abstract>
</article-meta>
</front>
</pmc>
</record>

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Detection of Brugia Parasite DNA in Human Blood by Real-Time PCR▿

Detection of Brugia Parasite DNA in Human Blood by Real-Time PCR▿

Source :

Abstract

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