A filter paper method for the detection of Plasmodium falciparum gametocytes by reverse transcription polymerase chain reaction.

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/microL. 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.

[1]  S. Nakazawa,et al.  Utility of the dried blood on filter paper as a source of cytokine mRNA for the analysis of immunoreactions in Plasmodium yoelii infection. , 2003, Acta tropica.

[2]  B. Hall,et al.  CD36 and intercellular adhesion molecule 1 mediate adhesion of developing Plasmodium falciparum gametocytes , 1996, Infection and immunity.

[3]  O. Doumbo,et al.  Pyrimethamine and proguanil resistance-conferring mutations in Plasmodium falciparum dihydrofolate reductase: polymerase chain reaction methods for surveillance in Africa. , 1995, The American journal of tropical medicine and hygiene.

[4]  T. Bousema,et al.  (Sub)microscopic Plasmodium falciparum gametocytaemia in Kenyan children after treatment with sulphadoxine-pyrimethamine monotherapy or in combination with artesunate. , 2006, International journal for parasitology.

[5]  J. Verhave,et al.  Age-dependent distribution of Plasmodium falciparum gametocytes quantified by Pfs25 real-time QT-NASBA in a cross-sectional study in Burkina Faso. , 2007, The American journal of tropical medicine and hygiene.

[6]  W. Trager,et al.  Human malaria parasites in continuous culture. , 1976, Science.

[7]  M. Smalley.  Plasmodium falciparum gametocytogenesis in vitro , 1976, Nature.

[8]  H. Babiker,et al.  Dynamics of gametocytes among Plasmodium falciparum clones in natural infections in an area of highly seasonal transmission. , 2002, The Journal of infectious diseases.

[9]  Petra Schneider,et al.  Quantification of Plasmodium falciparum gametocytes in differential stages of development by quantitative nucleic acid sequence-based amplification. , 2004, Molecular and biochemical parasitology.

[10]  J. Beeson,et al.  Selective Accumulation of Mature Asexual Stages of Plasmodium falciparum-Infected Erythrocytes in the Placenta , 2002, Infection and Immunity.

[11]  R. Carter,et al.  Detection of low level Plasmodium falciparum gametocytes using reverse transcriptase polymerase chain reaction. , 1999, Molecular and biochemical parasitology.