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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
Detection of Wuchereria bancrofti L3 larvae in mosquitoes: A reverse transcriptase PCR assay evaluating infection and infectivity
PLoS Neglected Tropical Diseases, Volume 4, No. 2, Article e602, Year 2010
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Description
Background: Detection of filarial DNA in mosquitoes by PCR cannot differentiate infective mosquitoes from infected mosquitoes. In order to evaluate transmission risk an assay is needed that can specifically detect infective L3 stage parasites. We now report the development of an assay that specifically detects the infective stage of Wuchereria bancrofti in mosquitoes. The assay detects an L3-activated mRNA transcript by reverse-transcriptase PCR (RT-PCR). Methodology/Principal Findings: W. bancrofti cuticle-related genes were selected using bioinformatics and screened as potential diagnostic target genes for L3 detection in mosquitoes. Expression profiles were determined using RT-PCR on RNA isolated from mosquitoes collected daily across a two-week period after feeding on infected blood. Conventional multiplex RT-PCR and real-time multiplex RT-PCR assays were developed using an L3-activated cuticlin transcript for L3 detection and a constitutively expressed transcript, tph-1, for 'any-stage' detection. Conclusions/Significance: This assay can be used to simultaneously detect W. bancrofti infective stage larvae and 'anystage' larvae in pooled vector mosquitoes. This test may be useful as a tool for assessing changes in transmission potential in the context of filariasis elimination programs. © 2010 Laney et al.
Available Materials
https://efashare.b-cdn.net/share/pmc/articles/PMC2821903/bin/pntd.0000602.s001.doc
Authors & Co-Authors
Laney, Sandra J.
United States, Northampton
Smith College
Egypt, Cairo
College of Science
Ramzy, Reda M.R.
Egypt, Cairo
Nutrition Institute
Helmy, Hanan H.
Egypt, Cairo
Ain Shams University
Farid, H. A.
Egypt, Cairo
Ain Shams University
Ashour, A. A.
Saudi Arabia, Taif
Taif University
Weil, Gary J.
United States, St. Louis
Washington University School of Medicine in St. Louis
Williams, Steven A.
United States, Northampton
Smith College
United States, Amherst
University of Massachusetts Amherst
Statistics
Citations: 33
Authors: 7
Affiliations: 7
Identifiers
Doi:
10.1371/journal.pntd.0000602
Research Areas
Genetics And Genomics