A qPCR-based multiplex assay for the detection of Wuchereria bancrofti, Plasmodium falciparum and Plasmodium vivax DNA

Ramakrishna U. Rao, Yuefang Huang, Moses J. Bockarie, Melinda Susapu, Sandra J. Laney, Gary J. Weil

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

The purpose of this study was to develop real-time multiplex quantitative PCR (qPCR) assays for the simultaneous detection of Wuchereria bancrofti (Wb), Plasmodium falciparum (Pf) and P. vivax (Pv) in mosquitoes. We optimized the assays with purified DNA samples and then used these assays to test DNA samples isolated from Anopheles punctulatus mosquitoes collected in villages in Papua New Guinea where these infections are co-endemic. Singleplex assays detected Wb, Pf and Pv DNA in 32%, 19% and 15% of the mosquito pools, respectively, either alone or together with other parasites. Multiplex assay results agreed with singleplex results in most cases. Overall parasite DNA rates in mosquitoes, estimated by PoolScreen 2 software, for Wb, Pf and Pv were 4.9%, 2.7% and 2.1%, respectively. Parasite DNA rates were consistently higher in blood-fed mosquitoes than in host-seeking mosquitoes. Our results show that multiplex qPCR can be used to detect and estimate prevalence rates for multiple parasite species in arthropod vectors. We believe that multiplex molecular xenodiagnosis has great potential as a tool for non-invasively assessing the distribution and prevalence of vector-borne pathogens such as W. bancrofti and Plasmodium spp. in human populations and for assessing the impact of interventions aimed at controlling or eliminating these diseases.

Original languageEnglish
Pages (from-to)365-370
Number of pages6
JournalTransactions of the Royal Society of Tropical Medicine and Hygiene
Volume103
Issue number4
DOIs
StatePublished - Apr 2009

Keywords

  • Mosquitoes
  • Plasmodium falciparum
  • Plasmodium vivax
  • Real-time PCR
  • Wuchereria bancrofti
  • Xenomonitoring

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