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Am. J. Trop. Med. Hyg., 78(1), 2008, pp. 114-116
Copyright © 2008 by The American Society of Tropical Medicine and Hygiene

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SHORT REPORT


A Filter Paper Method for the Detection of Plasmodium falciparum Gametocytes by Reverse Transcription–Polymerase Chain Reaction

Godfree Mlambo, Yessika Vasquez, Ralph LeBlanc, David Sullivan, AND Nirbhay Kumar*
Malaria Research Institute, Department of Molecular Microbiology and Immunology, Johns Hopkins University, Bloomberg School of Public Health, Baltimore, Maryland

 

ABSTRACT

Plasmodium falciparum gametocytes are obligate parasite sexual stages required for transmission of malaria from human hosts to the mosquito vector. Assessment of gametocyte carriers in the population is critical in understanding malaria transmission dynamics and in epidemiology studies. We applied a reverse transcription–polymerase chain reaction (RT-PCR)–based approach to detect pfs25 transcripts from blood dried on different filter papers in the laboratory. The detection limit was 1–2 gametocytes/µL. We further validated this assay by analyzing RNA in 10 matched blood samples (liquid blood and blood spotted on filter papers) collected from subjects under field conditions in Zambia. These results thus establish feasibility of detection of Plasmodium falciparum gametocytes by RT-PCR method from dried blood on filter paper. This assay will greatly facilitate bulk analysis of gametocyte RNA transcripts on filter paper, especially in areas where collection and preservation of liquid blood is not feasible.

Received July 9, 2007. Accepted for publication September 30, 2007.

Acknowledgments: We thank the Johns Hopkins Malaria Research Institute core facility for P. falciparum gametocyte cultures and Phil Thuma for facilitating Zambian field sample collection.

Financial support: Samples from Zambia were collected for a pilot project funded by the Johns Hopkins Malaria Research Institute under an approved protocol. Research in the laboratory of Nirbhay Kumar is supported by grants from the National Institutes of Health (NIH). The supply of human erythrocytes RBC is supported by NIH grant RR00052.

* Address correspondence to Nirbhay Kumar, Malaria Research Institute, Department of Molecular Microbiology and Immunology, Johns Hopkins University, Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, MD 21205. E-mail: nkumar{at}jhsph.edu

Authors’ address: Godfree Mlambo, Yessika Vasquez, Ralph Le-Blanc, David Sullivan and Nirbhay Kumar, Malaria Research Institute, Department of Molecular Microbiology and Immunology, Johns Hopkins University, Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, MD 21205.

Reprint requests: Nirbhay Kumar, Malaria Research Institute, Department of Molecular Microbiology and Immunology, Johns Hopkins University, Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, MD 21205, Telephone: 410-955-7177, Fax: 410-955-0105, E-mail: nkumar{at}jhsph.edu.






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Copyright © 2008 by the American Society of Tropical Medicine and Hygiene.