Mlambo Godfree, Vasquez Yessika, LeBlanc Ralph, Sullivan David, Kumar Nirbhay
Malaria Research Institute, Department of Molecular Microbiology and Immunology, Johns Hopkins University, Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, MD 21205, USA.
Am J Trop Med Hyg. 2008 Jan;78(1):114-6.
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.
恶性疟原虫配子体是疟疾从人类宿主传播至蚊媒所必需的专性寄生虫有性阶段。评估人群中的配子体携带者对于理解疟疾传播动态以及流行病学研究至关重要。我们应用基于逆转录-聚合酶链反应（RT-PCR）的方法在实验室中检测不同滤纸上干燥血液中的pfs25转录本。检测限为1-2个配子体/微升。我们通过分析从赞比亚野外条件下采集的10份配对血液样本（液态血和滤纸上的血斑）中的RNA进一步验证了该检测方法。因此，这些结果确立了通过RT-PCR方法从滤纸上的干燥血液中检测恶性疟原虫配子体的可行性。该检测方法将极大地促进对滤纸上配子体RNA转录本的批量分析，尤其是在无法采集和保存液态血的地区。
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