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We developed and evaluated real-time polymerase chain reaction (PCR) assays for detecting Wuchereria bancrofti DNA in human blood and in mosquitoes. An assay based on detection of the W. bancrofti "LDR" repeat DNA sequence was more sensitive than an assay for Wolbachia 16S rDNA. The LDR-based assay was sensitive for detecting microfilarial DNA on dried membrane filters or on filter paper. We also compared real-time PCR with conventional PCR (C-PCR) for detecting W. bancrofti DNA in mosquito samples collected in endemic areas in Egypt and Papua New Guinea. Although the two methods had comparable sensitivity for detecting filarial DNA in reference samples, real-time PCR was more sensitive than C-PCR in practice with field samples. Other advantages of real-time PCR include its high-throughput capacity and decreased risk of cross-contamination between test samples. We believe that real-time PCR has great potential as a tool for monitoring progress in large-scale filariasis elimination programs.
Received July 27, 2005. Accepted for publication December 8, 2005.
Acknowledgments: The authors would like to acknowledge the efforts of field teams and technical staff at Ain Shams University, Cairo, Egypt, and at the Papua New Guinea Institute of Medical Research in Madang. We thank K. Curtis for technical help.
Financial support: This work was supported by National Institutes of Health Grant AI 35855.
* Address correspondence to R. U. Rao, Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, Box 8051, 660 S Euclid Avenue, St. Louis, MO 63110. E-mail: rrao{at}im.wustl.edu
Authors’ addresses: Ramakrishna U. Rao, Laura J. Atkinson, and Gary J. Weil, Infectious Diseases Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO 63110, E-mails: rrao{at}wustl.edu, gweil{at}im.wustl.edu, and lja836{at}yahoo.com. Reda M. R. Ramzy, Hanan Helmy, and Hoda A. Farid, Research & Training Center on Vectors of Diseases, Ain Shams University, Abbassia, Cairo 11566, Egypt, E-mails: reda_m{at}masrawy.com, helmy26_2000{at}yahoo.com, and hafarid{at}rtcasuegypt.org. Moses J. Bockarie, Papua New Guinea Institute of Medical Research, Madang, Papua New Guinea, and the Center for Global Health and Diseases, Case-Western Reserve University, Cleveland, OH 44106, E-mail: moses.bockarie{at}case.edu. Melinda Susapu, Papua New Guinea Institute of Medical Research, Madang, Papua New Guinea, E-mail: msusapu{at}datec.net.pg. Sandra J. Laney and Steven A. Williams, Department of Biological Sciences, Smith College, Northampton, MA 01063. E-mails: slaney{at}smith.edu and swilliam{at}smith.edu.
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