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PCR and Mosquito dissection as tools to monitor filarial infection levels following mass treatment

David S Goodman1, Jean-Nicolas Orelus2, Jacquelin M Roberts1, Patrick J Lammie1 and Thomas G Streit3

Centers for Disease Control and Prevention, Division of Parasitic Diseases, Atlanta, Georgia, USA

Hopital St. Croix, Leogane, Haiti

University of Notre Dame, Center for Tropical Disease Research & Training, Notre Dame, Indiana, USA

Filaria Journal 2003, 2:11doi:10.1186/1475-2883-2-11

Published: 7 July 2003

Abstract

Background

Entomological methods may provide important tools for monitoring the progress of lymphatic filariasis elimination programs. In this study, we compared dissection of the vector, Culex quinquefasciatus, with the polymerase chain reaction (PCR) to assess filarial infection levels in mosquitoes in the context of a lymphatic filariasis elimination program in Leogane, Haiti.

Methods

Mosquitoes were collected using gravid traps located in 4 sentinel communities with Wuchereria bancrofti microfilaria prevalence that ranged from 0.8% to 15.9%. Captured mosquitoes were divided between dissection, to enumerate W. bancrofti larvae (L1, L2, L3) and desiccation for later analysis by PCR. PCR was conducted on DNA extracts from pooled mosquitoes (1–15 pooled females) utilizing a competitive PCR system with primers specific for the Ssp I repeat. PCR products were analyzed with a hybridization ELISA using probes specific for a control sequence and the Ssp I repeat.

Results

The prevalence of mosquito infection with W. bancrofti ranged from 0%–3.66% by dissection (L1–L3) and point estimates of infection prevalence, as assayed by PCR, ranged from 0.25% – 9.16%. Following mass treatment, W. bancrofti infection prevalence dropped significantly as determined by PCR and dissection in 2 of the 4 sentinel sites (Leogane and Barrier Jeudi, P = 0.04 and P = 0.005, respectively). Although transmission declined in the other two sites, larval recoveries were low and these changes were not statistically significant.

Discussion

Our results suggest that a single round of mass treatment can have an impact on transmission of lymphatic filariasis. The use of entomologic methods as a tool to monitor filariasis programs and the statistical limitations of mosquito trapping are discussed.


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